Common Species Research Articles

The use of earwax to determine livestock exposure to teratogenic lupine

Lupines (Lupinus spp.) are a common plant species on western U.S. rangelands with several lupine species containing alkaloids that can be toxic and/or teratogenic to livestock. In North America, more than 150 lupine species are recognized with some ranches or grazing allotments containing multiple species. One or more of these lupine species may contain alkaloids that are teratogenic to cattle. Previous work has shown that lupine alkaloids can be detected in earwax of cattle grazing lupine infested rangelands. Our hypothesis is that earwax can be used to determine if cattle have been exposed to teratogenic alkaloids from multiple lupine species. Two lupine species, L. sericeus and L. polyphyllus, were present on a rangeland in east-central Idaho. The teratogen, anagyrine, was detected in L. sericeus and the teratogen, ammodendrine, was detected in L. polyphyllus plants collected on this rangeland. In this study, earwax was collected from 69 pregnant cows that had previously grazed a rangeland containing two different lupine species containing alkaloids that cause crooked calf syndrome (CCS). Anagyrine was detected in the earwax of all 69 cows sampled. Ammodendrine, was detected in the earwax of 28 of the 69 cows sampled. Earwax is a good non-invasive sample to aid in the diagnosis of cattle that have consumed lupine and does appear, in this case, to be a good diagnostic tool to differentiate between more than one lupine species that may be the cause of CCS. Concentrations of anagyrine or ammodendrine did not correlate with the incidence of CCS.

Effect of air pollution on the leaf morphology and anatomy of common plant species of Kathmandu Valley

Effect of air pollution on the leaf morphology and anatomy of common plant species of Kathmandu Valley

Mechanisms of coexistence: Exploring species sorting and character displacement in woody plants to alleviate belowground competition.

Rarely do we observe competitive exclusion within plant communities, even though plants compete for a limited pool of resources. Thus, our understanding of the mechanisms sustaining plant biodiversity might be limited. In this study, we explore two common ecological strategies, species sorting and character displacement, that promote coexistence by reducing competition. We assess the degree to which woody plants may implement these two strategies to lower belowground competition for nutrients which occurs via nutritional (mostly mycorrhizal) mutualisms. First, we compile data on plant traits and the mycorrhizal association state of woody angiosperms using a global inventory of indigenous flora. Our analysis reveals that species in locations with high mycorrhizal diversity exhibit distinct mean values in leaf area and wood density based on their mycorrhizal type, indicating species sorting. Second, we reanalyse a large dataset on leaf area to demonstrate that in areas with high mycorrhizal diversity, trees maintain divergent leaf area values, showcasing character displacement. Character displacement among plants is considered rare, making our observation significant. In summary, our study uncovers a rare occurrence of character displacement and identifies a common mechanism employed by plants to alleviate competition, shedding light on the complexities of plant coexistence in diverse ecosystems.

An exploratory analysis of the development and implementation of urban animal welfare policy in public spaces for four common species: Cats, dogs, pigeons, and foxes in the Brussels Capital Region

ABSTRACT This study aims to understand urban animal welfare policy development and implementation for four species in public spaces: cats (Felis catus), dogs (Canis familiaris), pigeons (Columbia livia domestica), and foxes (Vulpes vulpes). Our exploratory research offers an overview of the perspectives of all involved municipal and police officers and their challenges in a metropolitan urban context, the Brussels Capital Region in Belgium. We conducted semi-structured interviews with 35 participants from 19 municipalities and six police zones of the region. Additionally, we organized two focus groups consisting of representatives from municipalities, police zones, and nongovernmental organizations. Afterward, we analyzed the data thematically, leading to the identification of six policy and six practice recommendations. These recommendations can help advance the notion of urban animal welfare for the four species from a multispecies perspective.

Algal Biodiversity of Nine Megaliths in South-East Bulgaria.

This paper presents the first data on the biodiversity of lithophytic algae from Bulgarian megaliths obtained after the application of the direct sampling method, subsequent cultivation, and processing by light microscopy. A rich algal flora was found: 90 species and 1 variety of 65 genera from Cyanoprokaryota/Cyanobacteria (29 species, 13 genera), Chlorophyta (40 species and 1 variety, 38 genera), Streptophyta (5 species, 1 genus), and Ochrophyta (16 species, 13 genera). Among them were the globally rare Pseudodictyochloris multinucleata (Chlorophyta), found for the first time in such lowland and warm habitats, and Scotiella tuberculata (Chlorophyta), for which this is the first finding in the country. Three of the recorded species are conservationally important. The low floristic similarity between the sites (0-33%) shows the diversity of the algal flora, with no common species found for all the megaliths studied. The most widespread were the strongly adaptive and competitive Stichococcus bacillaris, Apatococcus lobatus, and Chloroidium ellipsoidium (Chlorophyta). The correlations estimated between the species number and substrate temperature (18.1-49.6 °C) suggest the prospect of future research related to the impact of global warming. In addition, the study points to the safety aspects as it revealed species from nine potentially toxin-producing cyanoprokaryotic genera that could be harmful to visitors' health.

Antimicrobial Activity of Bark from Four North American Tree Species.

Although many backcountry first aid kits contain antibiotic ointment, the supply can be quickly exhausted if a patient has extensive wounds or if there are multiple patients. We assessed the antibacterial properties of bark extract from four North American woody plant species known to native Missourians as medicinal plants (Quercus macrocarpa, Salix humilis, Pinus echinata, and Hamamelis vernalis). We tested their antimicrobial properties, with the disc diffusion technique, against four common pathogenic bacterial species: Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterobacter aerogenes (now known as Klebsiella aerogenes). We report evidence of antibacterial activity of bark extract from all four plant species. Our results confirm that traditional uses of these species may be useful in fighting infection and could be especially useful in a wilderness setting when modern antibiotics are exhausted.

Exploring the interspecific relationships among Dendrobium species via electrochemical fingerprinting techniques

This study investigated the interspecific relationships among 29 Dendrobium species using electrochemical fingerprinting techniques. Differential pulse voltammetry was employed to record species-specific electrochemical profiles in both phosphate buffer solution (PBS) and acetic acid buffer solution (ABS). Distinct voltammetric patterns were observed for each species, with ABS yielding higher peak currents compared to PBS. On average, peak currents in ABS were 37.2 % ± 5.8 % higher than in PBS across all species. For example, D. nobile showed a peak current of 12.3 μA in ABS compared to 8.9 μA in PBS, a 38.2 % increase. Normalized two-dimensional plots of the electrochemical fingerprints served as unique signatures for species differentiation. Cluster analysis of the electrochemical data revealed phylogenetic relationships largely consistent with previous molecular studies, while also highlighting novel groupings. For instance, D. parishii and D. pulchellum formed separate branches, indicating distant relationships with other species. Common species such as D. officinale, D. nobile, D. primulinum, and D. loddigesii clustered together, confirming their close relationships. Interestingly, species from Sect. Dendrobium showed mixed clustering with species from four other sections, reflecting the genus's complex evolutionary history. While some inconsistencies with previous molecular phylogenetic studies were noted, such as the distant relationship between D. parishii and D. anosmum, the overall congruence between electrochemical and molecular data validates the potential of this approach. This study demonstrates that electrochemical fingerprinting, combined with appropriate data analysis techniques, can provide valuable insights into the taxonomic relationships and chemical diversity of Dendrobium species, offering a complementary tool to traditional morphological and molecular methods for plant systematics and conservation efforts.

Comparative study on metabolite variations of two rose teas by plant metabolomics and revealing their skin-whitening candidates by spectrum-effect relationship analysis.

Rosa rugosa var. plena Rehd (CBR) and Rosa chinensis cv. "JinBian" (JBR) are two common species used in rose tea among different original species. CBR, the officially documented original plant of the rose species for food and medicinal purposes, is more costly than JBR. With increasing demand for different rose teas, it is meaningful to compare the chemical constituents for their quality control and reveal their skin-whitening components that will provide in-depth insights for the expansion of the rose tea industry. This study aims to reveal the chemical variances between CBR and JBR and determine their skin-whitening components. A strategy obtained by combining MS-based plant-metabolomics with spectrum-effect relationship analysis has been proposed for unveiling chemical differences between CBR and JBR and further exploring their potential skin-whitening components. A total of 2030 metabolites were found that revealed considerable differences between CBR and JBR. The results of bioactivity assay demonstrated that JBR exhibited stronger tyrosinase inhibition activity than CBR. Six potential skin-whitening compounds (di-O-galloyl-HHDP-glucoside, tri-O-galloyl-HHDP-glucoside, spiraeoside, quinic acid, rugosin A, and 1,2,3,6-tetra-O-galloyl-glucose) were discovered as potential tyrosinase inhibitors, via spectrum-effect relationship analysis. This is the first time that di-O-galloyl-HHDP-glucoside, tri-O-galloyl-HHDP-glucoside, rugosin A, and 1,2,3,6-tetra-O-galloyl-glucose have been reported with tyrosinase inhibitory activity. Additionally, molecular docking analysis was used to reveal the inhibition mechanism of these compounds toward tyrosinase. The finding of this study will be of great importance for the quality control of the two types of rose teas, and the revealed active ingredients will provide in-depth insights for the expansion of the rose tea industry.

Biofilm Formation and Antibiotic Resistance Profiles in Carbapenemase-Producing Gram-Negative Rods-A Comparative Analysis between Screening and Pathological Isolates.

(1) Background: Carbapenem-resistant (CR) bacteria pose a significant global public health challenge due to their ability to evade treatment with beta-lactam antibiotics, including carbapenems. This study investigates the biofilm-forming capabilities of CR clinical bacterial isolates and examines the impact of serum on biofilm formation. Additionally, the study evaluates the resistance profiles and genetic markers for carbapenemase production. (2) Methods: Bacterial isolates were collected from the microbiology laboratory of Mures County Clinical Hospital between October 2022 and September 2023. Pharyngeal and rectal swabs were screened for carbapenem-resistant bacteria using selective media. Lower respiratory tract samples were also analyzed for CR Gram-negative bacteria. The isolates were tested for their ability to form biofilms in the presence and absence of fetal bovine serum at 24 and 48 h. Carbapenemase production was detected phenotypically and confirmed via PCR for relevant genes. (3) Results: Out of 846 screened samples, 4.25% from pharyngeal swabs and 6.38% from rectal swabs tested positive for CR bacteria. Acinetobacter baumannii and Klebsiella pneumoniae were the most common species isolated. Biofilm formation varied significantly between clinical isolates and standard strains, with clinical isolates generally showing higher biofilm production. The presence of serum had no significant effect on biofilm formation in Klebsiella spp., but stimulated biofilm formation for Acinetobacter spp. Carbapenemase genes blaKPC, blaOXA-48-like, and blaNDM were detected in various isolates, predominantly in Klebsiella spp., but were not the main determinants of carbapenem resistance, at least in screening isolates. (4) Conclusions: This study highlights the variability in biofilm formation among CR clinical isolates and underscores the differences between the bacteria found as carriage versus infection. Both bacterial species and environmental factors variably influence biofilm formation. These insights are crucial for the development of effective treatment and infection control strategies in clinical settings.

Habitat suitability modelling to predict the distribution of deep coral ecosystems: The case of Linosa Island (southern Mediterranean Sea, Italy)

In areas with limited field data, predictive habitat mapping is a valuable method for elucidating species-environment relationships and enhancing our knowledge of the spatial distribution and complexity of benthic habitats. Species distribution models (SDMs) can be an important tool to support in science-based ecosystem management. The availability of direct observations of mesophotic species, including gorgonians and black corals, during costly surveys is generally limited. Therefore, predicting the distribution of mesophotic species in relation to key physical parameters of the seafloor would help improving conservation strategies in existing and new Marine Protected Areas (MPAs). This study aims to assess the distribution of gorgonians and black corals off Linosa Island, in the Strait of Sicily, a biogeographic boundary area between the western and eastern Mediterranean. The volcanic island of Linosa represents a small, naturally preserved area, with very limited human pressure, hosting rich marine benthic biodiversity on its wide submarine portions. Distribution of the most common coral species off Linosa Island was modelled under an SDM framework, relying on direct observations collected during two research cruises in 2016 and 2017 and a series of terrain parameters acquired through geophysical techniques. We used the so-called “ensemble of small models” approach to calibrate SDMs, which achieved fair-to-excellent results (AUC >0.7). In addition to identifying depth as the primary factor influencing coral distribution, our study also highlighted ruggedness as a significant terrain variable. Specifically, the depth range of 110–230 m emerged as the critical parameter determining habitat suitability for all modelled species, also highlighting peculiar and specie–specific habitat requirements.

Quantitative assessment of reef foraminifera community from metabarcoding data.

Describing living community compositions is essential to monitor ecosystems in a rapidly changing world, but it is challenging to produce fast and accurate depiction of ecosystems due to methodological limitations. Morphological methods provide absolute abundances with limited throughput, whereas metabarcoding provides relative abundances of genes that may not correctly represent living communities from environmental DNA assessed with morphological methods. However, it has the potential to deliver fast descriptions of living communities provided that it is interpreted with validated species-specific calibrations and reference databases. Here, we developed a quantitative approach to retrieve from metabarcoding data the assemblages of living large benthic foraminifera (LBF), photosymbiotic calcifying protists, from Indonesian coral reefs that are under increasing anthropogenic pressure. To depict the diversity, we calculated taxon-specific correction factors to reduce biological biases by comparing surface area, biovolume and calcite volume, and the number of mitochondrial gene copies in seven common LBF species. To validate the approach, we compared calibrated datasets of morphological communities from mock samples with bulk reef sediment; both sample types were metabarcoded. The calibration of the data significantly improved the estimations of genus relative abundance, with a difference of ±5% on average, allowing for comparison of past morphological datasets with future molecular ones. Our results also highlight the application of our quantitative approach to support reef monitoring operations by capturing fine-scale processes, such as seasonal and pollution-driven dynamics, that require high-throughput sampling treatment.

The rising incidence of feline and cat-transmitted sporotrichosis in Latin America.

Sporotrichosis is a subcutaneous mycosis caused by different species of thermodimorphic fungi belonging to the genus Sporothrix. These fungi may affect a wide range of warm-blooded animals, including humans, dogs and, mainly, cats. In this article, we conducted a narrative literature review about the status of feline and cat-transmitted sporotrichosis in Latin American countries, with emphasis on Brazil. From a global perspective, Latin America is one of the main foci of human and feline sporotrichosis. In humans, the disease has been reported in several countries, with endemic transmission occurring in Brazil, Colombia, Costa Rica, Guatemala, Mexico and Uruguay. Sporothrix brasiliensis is the most common species identified in human cases in Brazil, whereas Sporothrix schenckii predominates in other countries. Domestic cats are the main source of infection in the zoonotic transmission cycle of sporotrichosis in Latin America. Cases of feline sporotrichosis have been reported in several countries, including Argentina, Brazil, Chile, Colombia, Mexico, Panama, Paraguay and Peru.

Invasive shrubs differentially alter autumnal activity for three common small-mammal species.

Seasonal variation in animal activity influences fitness and the intensity of ecological interactions (e.g., competition, predation), yet aspects of global change in the Anthropocene may catalyze shifts in seasonal activity. Invasive plants are components of global change and can modify animal daily activity, but their influence on animal seasonal activity is less understood. We examined how invasive woody shrubs (Autumn olive [Elaeagnus umbellata] and Amur honeysuckle [Lonicera maackii]) affect seasonal activity of three common small-mammal species by coupling experimental shrub removal with autumnal camera trapping for two consecutive years at six paired forest sites (total 12 plots). Eastern chipmunks (Tamias striatus) foraged more, and foraging was observed at least 20 days longer, in shrub-invaded forests. White-footed mice (Peromyscus leucopus) foraged more in invaded than cleared plots in one study year, but P. leucopus autumn activity timing did not differ between shrub-removal treatments. Fox squirrel (Sciurus niger) activity displayed year-specific responses to shrub removal suggesting intraannual cues (e.g., temperature) structure S. niger autumnal activity. Our work highlights how plant invasions can have species-specific effects on seasonal animal activity, may modify the timing of physiological processes (e.g., torpor), and could generate variation in animal-mediated interactions such as seed dispersal or granivory.

Relationship between Endotoxin Content in Vaccine Preclinical Formulations and Animal Welfare: An Extensive Study on Historical Data to Set an Informed Threshold.

The most widely known pyrogen impurity in vaccines is the Gram-negative bacterial endotoxin lipopolysaccharide (LPS). When administered at toxic doses, endotoxin triggers inflammatory responses, which lead to endotoxic shock. The literature on endotoxic content (EC) for preclinical vaccines' formulations used in animal studies is very poor, and the recommended thresholds are solely based on commercial vaccine limits set for humans and are, therefore, not connected to the actual impact of EC on animal welfare for species used in preclinical research studies. An extensive study to evaluate the presence of a potential relationship between endotoxin content in formulations administered to mice (the most common species used in preclinical research studies) and their welfare was conducted to calculate an EC threshold for formulations of candidate vaccines. Three years of historical data, from more than 500 formulations of different antigen types (i.e., proteins, glycoconjugates, OMV/GMMA) injected into more than 5000 mice, was evaluated with two alternative statistical methodologies, both demonstrating that there is no significant relationship between actual endotoxin levels and mouse welfare. The calculation of thresholds was, therefore, performed by consistency versus formulations that demonstrated no impact on animal welfare.

Acaricidal activity of Egyptian crude plant extracts against Haemaphysalis longicornis ticks.

Haemaphysalis longicornis is a common Ixodida tick species found in temperate areas of Asian countries. An anti-tick assay was conducted on adult female H. longicornis ticks. Plant extract solutions were prepared at concentrations of 50, 25, and 10 mg/mL. Tick survival and mortality were assessed by counting the number of dead and live ticks at 24 h, 48 h, 72 h, and 96 h posttreatment. Out of 11 plant extracts screened, Artemisia judaica extract exhibited the highest potency with 100% mortality (5/5) at 48 h when applied at high and moderate concentrations (50 and 25 mg/mL). Similar results were observed at 96 h for the 10 mg/mL group compared to the untreated ticks. Cleome droserifolia extract demonstrated partial activity with 60% (3/5) and 20% (1/5) mortality at 96 h posttreatment at concentrations of 50 and 25 mg/mL, respectively. Forsskaolea tenacissima extract showed a weak effect with 100% tick mortality (5/5) only at the highest treatment concentration after 96 h. To confirm the activity of A. judaica, trial 2 was conducted. A. judaica demonstrated potency within 48 h in high dose and 72 h in moderate dose, with 100% mortality (15/15) at 96 h posttreatment compared to untreated ticks. The median lethal time 50 (LT50) values were 30.37 h for the high and 55.08 h for the moderate doses. Liquid chromatography‒mass spectrometry was performed on the most potent candidate (A. judaica) to identify its phytochemical components. The results revealed the presence of 9 compounds identified through manual annotation and 74 compounds from the Global Natural Products Social library. These compounds included terpenoids, steroids, phenylpropanoids, flavonoid glycosides, flavonoids, and benzenoids. Camphor was identified as the major component via both approaches. These findings suggest the potential use of A. judaica extract in the future development of acaricidal therapeutics.

Lateral connectivity maintains higher freshwater mussel biodiversity

Abstract The loss of lateral hydrological connectivity (LLHC) in freshwater ecosystems is increasingly threatening biodiversity; however, studies reporting this situation are limited, especially those assessing multi‐measure biodiversity features at large spatial scales. Here, and using one the most threatened taxonomic groups in freshwater ecosystems, we examined the possible influence of LLHC on the species diversity, density, and biomass of mussel assemblages and the genetic diversity and structure of Nodularia douglasiae (a common and widespread native species) in the Yangtze River floodplain. Results showed that the species diversity of freshwater mussels (α and β diversity) significantly differed between the connected and the disconnected areas, with higher α diversity and lower β diversity in the connected areas. At the same time, relative abundance, density, biomass, and genetic diversity of freshwater mussels were higher in the connected areas. High genetic differentiation and low gene flow were found among the 18 analysed N. douglasiae populations, indicating that LLHC is affecting genetic structure of this particular freshwater mussel species. Overall, results indicated that river–lake connectivity is affecting freshwater mussel biodiversity. In addition, this study highlights that multi‐measure biodiversity features provide distinct information about biodiversity dynamics, and are essential to evaluate the effects of floodplain disconnection. We advocate that an integrative approach to floodplain management embracing species and genetic diversity is needed for effective biodiversity conservation in large river ecosystems.

Regeneration of post‐mining and post‐fire soil function by assessment of tree nutrient status: Evidence from pioneer and N‐fixing species

AbstractNutrient concentrations in the foliage of particular tree species can identify growth‐limiting factors. Therefore, the adaptability and usefulness of certain trees in the afforestation of barren soils after disturbances could be based on nutrient supply and stoichiometry. In this study, we investigated the macronutrient supply to (i.e., nitrogen [N], phosphorus [P], potassium, sulfur, calcium [Ca], and magnesium [Mg]) and C:N:P (carbon:nitrogen:phosphorus) stoichiometry of the foliage of black alder (Alnus glutinosa), common birch (Betula pendula), and Scots pine (Pinus sylvestris) in two different regeneration scenarios—a reclaimed mine site with primary succession and an afforested post‐fire (PF) site with secondary succession. The control plot was on an undisturbed forest site hosting stands of the studied tree species. The lower concentrations of elements in the post‐mining soils developed from “point zero” were enough to supply the trees with a similar level of nutrition to those in the PF and undisturbed soils. The studied tree species differed in their foliage chemistry and had different effects on the soil properties. The alder and birch foliage contained more nutrients than the pine foliage. In particular, the birch foliage contained the highest concentrations of Ca, Mg, and P, thus enhancing the biogeochemical cycling of these elements. The alder foliage had the highest N concentration, albeit the soil total N contents under this species were not higher than those under birch. Thus, its phytomeliorative effect may have been overestimated. The pine foliage contained the lowest concentrations of nutrients, indicating the high nutrient use efficiency of this species. On the sites degraded after disturbance, pine and birch grew under N‐limitation stress, while the alder—a N‐fixing species—grew under P‐limitation stress, especially at the PF site. The PF soils under alder were also characterized by higher acidity, P depletion, and higher C:P and N:P ratios compared to the PF soils under the other studied species. We found that birch should be the most common tree species in the first phase of area regeneration after disturbances. Introducing alder into afforestation should take place in admixtures, especially at the PF site.

Vertical and horizontal environmental DNA (eDNA) patterns of fish in a shallow and well-mixed North Sea area

The integration of eDNA metabarcoding into monitoring programs provides valuable information about fish community structures. Despite the growing body of evidence supporting the method's effectiveness in distinguishing fine-scale eDNA signals, there is a limited understanding of eDNA distribution in shallow, well-mixed environments, especially related to sampling depth. We analyzed 167 samples collected from the surface and bottom water at 17 locations of the Belgian Part of the North Sea (BPNS), where the deepest sampling point was 31 m, and compared this to beam trawl catch data collected simultaneously at the same locations. eDNA metabarcoding identified an additional 22 species compared to beam trawl catch data. Diversity measures and patterns were very similar between surface and bottom samples and revealed community patterns that were previously described by long-term beam trawl catch data. Surface and bottom samples had 39 fish species in common, while six and eight rare species were uniquely detected, respectively. Our results demonstrate that eDNA metabarcoding effectively identifies spatial community patterns of fishes in the highly dynamic environment of the BPNS regardless of sampling depth. Out of the six most common species tested, eDNA metabarcoding read abundances correlated strongly with catch-based abundance data for one species, but moderately for two others, indicating that inferring fish abundance and biomass via eDNA metabarcoding remains challenging.

Ten lessons for rare wildlife species conservation when coming from a common species background

AbstractEfforts to slow biodiversity loss have ushered in an era of proactive conservation for rare species, or species that have any combination of low relative abundance, high habitat specificity, and limited geographic range. Approaches to wildlife management and conservation were often developed for more abundant, generalist species, and frequently do not translate well to rare species. Conservation for rare species therefore requires adapting knowledge, action and inquiry tools for common species to rare species. We therefore present 10 lessons for adapting conservation tools to rare species, each with advice to implement the lessons. Several themes emerge from these lessons. Information scarcity necessitates high standards of information for rare species. Missing rare species information can be complemented with multiple forms of knowledge, particularly when the need for conservation action precludes additional data collection. Investment in depth (over breadth) of information for individuals of rare species can provide alternatives to population trend, which has lower reliability for rare species. Natural history documentation can improve rare species conservation strategies. Finally, curiosity‐driven inquiry can inspire rare species conservation innovation. We hope providing guidance that addresses challenges for rare species will help others bridge the gap between common wildlife approaches and proactive rare species conservation.

Genetic diversity of Plasmodium malariae in sub-Saharan Africa: a two-marker genotyping approach for molecular epidemiological studies.

Plasmodium malariae is the most common non-falciparum species in sub-Saharan Africa. Despite this, data on its genetic diversity is scarce. Therefore, we aimed to establish a P. malariae genotyping approach based on size polymorphic regions that can be easily applied in molecular epidemiological studies. Four potential genotyping markers, Pm02, Pm09, P. malariae thrombospondin-related anonymous protein (pmtrap), and P. malariae merozoite surface protein fragment 2 (pmmsp1 F2) were amplified via nested PCR and analysed using automated capillary gel electrophoresis. We observed the highest allelic diversity for pmtrap (MOI = 1.61) and pmmsp1 F2 (He = 0.81). Further applying the two markers pmtrap and pmmsp1 F2 on a different sample set of 21 P. malariae positive individuals followed up over one week, we saw a high consistency in their performance. The results show a large complexity and high dynamics of P. malariae infections in the asymptomatic Gabonese study population. We successfully implemented a new genotyping panel for P. malariae consisting of only two markers: pmtrap and pmmsp1 F2. It can be easily applied in other settings to investigate the genotype diversity of P. malariae populations, providing further important data on the molecular epidemiology of this parasite species.