Colony Density Research Articles

A simple, robust, broadly applicable insertion mutagenesis method to create random fluorescent protein: target protein fusions.

A simple, broadly applicable method was developed using an in vitro transposition reaction followed by transformation into Escherichia coli and screening plates for fluorescent colonies. The transposition reaction catalyzes the random insertion of a fluorescent protein open reading frame into a target gene on a plasmid. The transposition reaction is employed directly in an E. coli transformation with no further procedures. Plating at high colony density yields fluorescent colonies. Plasmids purified from fluorescent colonies contain random, in-frame fusion proteins into the target gene. The plate screen also results in expressed, stable proteins. A large library of chimeric proteins was produced, which was useful for downstream research. The effect of using different fluorescent proteins was investigated as well as the dependence of the linker sequence between the target and fluorescent protein open reading frames. The utility and simplicity of the method were demonstrated by the fact that it has been employed in an undergraduate biology laboratory class without failure over dozens of class sections. This suggests that the method will be useful in high-impact research at small liberal arts colleges with limited resources. However, in-frame fusion proteins were obtained from 8 different targets suggesting that the method is broadly applicable in any research setting.

Demographic recovery of corals at a wave-exposed reef following catastrophic disturbance

Rapid recovery of coral cover following acute disturbance has been documented on many reefs. Yet measuring coverage alone can mask shifts in community and demographic structure. Here, we quantify long-term changes in population size structure for three common genera (Acropora, Pocillopora, and Stylophora) at an eastern outer reef in Palau, Micronesia, following catastrophic loss of corals due to typhoon Bopha in 2012, based on size measurements from 3648 coral colonies. Mean colony size returned to pre-disturbance levels within 4 and 6 years for Stylophora and Pocillopora, respectively. By 2020, Pocillopora colony density far exceeded pre-disturbance levels, with rapid successful recruitment following typhoon Bopha. Despite recovery of Acropora colony density by 2020, populations remained dominated by smaller colonies. We demonstrate that full demographic recovery can occur more rapidly for pocilloporids (within 6 years) compared to Acropora which had not fully recovered by 8 years post-disturbance, possibly due to fewer annual recruitment events and larger maximum colony sizes. Our results highlight the value of demographic metrics as early indicators of recovery.

Differential expression of antimicrobial peptide encoding genes of honey bee (Apis mellifera) after Varroa mite (Varroa destructor) infestation

Varroa destructor mite is a serious threat to the world apiculture industry, and severely hampered beekeeping practices. Innate immune systems are key defenses of animals and are particularly important in species that lack the sophisticated adaptive immune systems found in vertebrates. In honey bees, antimicrobial immune peptide genes play a significant immune response against pathogens. The present study analyzed the detrimental effects of Varroa infestation on Apis mellifera at all developmental stages in both Varroa challenged and controlled hives. Physical attributes were observed in Varroa hives at different time intervals (0, 30, and 45 days respectively). After 45 days of Varroa exposure, the samples were taken, and the AMPs immune response of bees parasitized by Varroa was detected by studying the expression levels of antibacterial peptides (defensin, apidaecin, abaecin, and hymenoptaecin). Significant alterations in the physical parameters viz. body length, body weight, wing shape, and mean numbers of brood cells were observed compared to the control. Colony density was also decreased with marked negative behavioural changes. Ct values of Real-Time PCR results showed that AMP-specific immune gene transcription level in Varroa-challenged bees was downregulated, except for abaecin and defensin (adult stage). There was a significant (p < 0.05) increase in abaecin gene transcription folds i.e., 1.064 larvae, 1.732 pupae, and 1.264 adults bees. As a whole, all these findings can be taken as critical biomarkers in the advancement of control techniques against Varroa and other pathogen infections as a part of wide-ranging programs of honey bee health and Varroa control.

The use of drone congregation behaviour for population surveys of the honey bee Apis cerana

Honey bees (Apis spp.) are important pollinators in many natural and agro-ecosystems across the world. Effective means of surveying wild populations are therefore key to their conservation and management. One available survey method infers honey bee colony density from the genotype of drones (males) sampled from sites known as Drone Congregation Areas (DCAs). While this approach has been commonly used for the Western honey bee (A. mellifera), its feasibility for other Apis species is unknown. Here, we investigate drone congregation behaviour in the Asian honey bee Apis cerana in north-east Australia and its suitability for inferring colony density. As this A. cerana population is invasive, surveys in this case can aid in monitoring the population’s growth and spread. Over 5 years, we identified 30 DCAs, many of which were stable across time. DCAs were sheltered areas beside tree-lines or openings in the forest canopy. A. cerana drones joined DCAs during 1–2-h afternoon intervals and could be sampled at heights of 2–24 m via adhesive lines attached to helium balloons carrying lures coated in queen pheromone. Drones were more likely to be present at a DCA as temperature increased, though abiotic factors did not predict overall drone abundance. Drones could be sampled in low numbers even where colony density was extremely low. Based on the genotyping and inferred sibship of drones sampled at DCAs between 2016 and 2021, we estimate population density in Australia’s A. cerana to be in the range 1.1–8.1 colonies/km2. This extrapolates to a total population size in the range 11,000–83,000 colonies, with more refined estimates requiring better knowledge of drone flight distance and the effect of habitat on colony density. We conclude that population surveys based on drones from DCAs are possible for A. cerana and propose that this approach be part of a toolkit of methods used to monitor Asian honey bee populations in both their native and invasive ranges.

INVASIVE INSECT SPECIES CYDALIMA PERSPECTALIS (WALKER, 1859): GLOBAL DISTRIBUTION, INVASION IN UKRAINE AND DAMAGE IN CHERNIVTSI REGION

Invasions of species into ecosystems cause their transformation and, in certain cases, their degradation. The study was carried out to determine the extent of the invasion of Cydalima perspectalis (Walker, 1859). For this purpose, the following aspects were analyzed: distribution of the species in different countries of the worldі, history of invasion of the pest in Ukraine, degree of damage to plantations of different box (Buxus L.) species and colony density in certain areas. The chronology of the distribution of C. perspectalis was studied using literature sources and international databases: Global Biodiversity Information Facility (GBIF), Delivering Alien Invasive Species Inventories for Europe (DAISIE), European and Mediterranean Plant Protection Organization (EPPO). The information on the history of the species' invasion in Ukraine and in particular in the Chernivtsi region was found in the literature, using the database of the National Biodiversity Information Network (UkrBIN), through questionnaires and personal observations using the route method (counting the number of C. perspectalis on a certain route). The degree of damage to Buxus sp. shrubs was determined visually using a scale. The density of C. perspectalis in each area was assessed by total counts and direct counts in situ with a parallel assessment of habitat size. It was found that the expansion of C. perspectalis beyond its native range began in 2006 from southwestern Germany. In 15 years, the species has occupied the territories of about 40 mainland and island countries of the North American and European continents. Since 2014, after the invasion of the Zakarpattia region, C. perspectalis has begun to spread throughout the Ukrainian territories. Currently, the secondary range of the species is represented by 14 regions. The presence of C. perspectalis has been detected in 16 settlements of Chernivtsi region, where the degree of damage to box shrubs is mostly severe. The density of caterpillars on most bushes is up to 100 individuals/m2, which indicates the high damage of the species. Key words: Cydalima perspectalis (Walker, 1859), invasive species, damage, expansion.

Gold Nanoparticle Coated Biocompatible Polymeric Matrix Mesh Plasticizers Embedded over Polyvinyl Chloride Plastic Porous Grid: Designed Reusable Microbial Screening Mask and Its Application

Under severe pandemic conditions, any airborne microbe, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), leads to increased fatality if physical screening is compromised. Therefore, an investigation was carried out to design a reusable screening mask that could promise the highest safety and be compatible with users. After going through the analytical and clinical investigation, the design and the work quality are confined to the ground version. To initiate top quality mask screener design of high-tech gold nanoparticles (AuNPs) hydroxyethyl methacrylate (HEMA) composite mask, safety evaluation data from all masks were collected. The porous polyvinyl chloride (PVC) circular grid frame was coated with a gold metallic nano particles layer, and the flexible adhesive polymeric matrix mesh plasticizers polyethylene terephthalate (PET) was used to coat bed on each grid, as a design. In-vitro quantitative analysis process assembly of bio-contaminant and specific matter was developed, in which breathed/vacuumed air was passed via fractional screening cone adhesive edged stumbling block route for final pure contaminant free air. The vacuumed air was collected through the adhesive slide, and microbial growth and particular matter were analysed and discovered to be best compared to conventional filter masks. Gold coat film entrapment tests with colony density were performed through Fourier transform infrared spectroscopy (FTIR), and virus crystal structure analysis was done through an X-ray diffractometer. Clinical investigations of this development were done with 50 volunteers. The research demonstrates the advantages of the AuNPs HEMA composite reusable mask design comparatively best over other mask designs regarding microbiological screening and lab safety, and it may be helpful to stand against biological warfare.

Densities of feral honey bee colonies utilising tree hollows in central Perth, Western Australia

Densities of feral honey bee colonies utilising tree hollows in central Perth, Western Australia

Variable prevalence of diseases and compromised health conditions on hard corals around Mauritius Island, Western Indian Ocean

Although coral diseases and compromised health conditions (CHCs) are one of the most prominent threats to coral reefs globally, they have yet to be thoroughly investigated in the Western Indian Ocean. This study aimed to assess the prevalence and spatial distribution of coral diseases and CHCs around Mauritius. Six coral diseases [Growth Anomaly (GA), Skeletal Eroding Band (SEB), Brown Band (BrBD), Porites White Patch Syndrome (PWPS), White Syndromes (WSs), and Yellow Band (YB)] and four CHCs [Pink Patches (PP), Pink Spots (PS), Pink Line Syndrome (PLS), and Focal Bleaching (FB)] were spatially variable in prevalence among the 17 studied sites. The average prevalence for Mauritius was 4.88% (SD 0.52%), almost half of what has been documented from Caribbean and Pacific reefs (about 10%). Total coral colony density (R = −0.585, P &lt; 0.001) and diseases/CHCs-affected host density (R = −0.249, P &gt; 0.05) were negatively correlated while human population density (R = 0.085, P &lt; 0.05) and coral cover (R = 0.045, P &lt; 0.05) were positively correlated with coral diseases/CHCs prevalence. The prevalence of diseases/CHCs significantly differed between the reef-flat and fore-reef zones, with the latter exhibiting higher prevalence. Cluster analyses revealed that PLS, YB, and FB were mostly distributed in the fore-reef zone, while the other diseases/CHCs were mostly predominant on the reef-flats. However, coral disease/CHCs prevalence did not differ significantly (F = 2.613, P &gt; 0.05) between protected and nonprotected zones. These findings have important implications for informing appropriate coral reef conservation and management strategies for Mauritius.

Effects on local oxygen conditions by the invasive macrophyte Myriophyllum spicatum

Myriophyllum spicatum, Eurasian watermilfoil, is a submerged aquatic plant invasive to North America. Several characteristics found in M. spicatum provide reasoning behind its invasion success such as its ability to spread and grow rapidly as well as displace other surrounding native species. However, Eurasian watermilfoil’s effects on ecosystem functioning (such as dissolved oxygen) and how such functioning differ from effects of native vegetation have seldom been studied. Using data collected in field, we used statistical models including Gaussian multivariate linear effect models and structural equation modelling (SEM), to investigate the effect of vegetation type and cover on dissolved oxygen (DO) and temperature gradients. Here, we show that invasive Eurasian watermilfoil colonies, relative to native submerged vegetation, can have a direct effect on DO gradients. These changes in DO conditions were driven by both an increase in surface oxygen concentrations and a decrease in bottom layer oxygen concentration in dense M. spicatum vegetation. Furthermore, we find that the differences in DO gradients could be predicted from M. spicatum’s direct impact on oxygen concentration and not indirectly via its effects on water temperature. Our results demonstrate that dense colonies of M. spicatum can directly affect DO concentrations and may do so more than native macrophytes which could explain its rapid spread and potential impacts on ecosystem functioning.

A pore-scale reactive transport modeling study for quorum sensing-driven biofilm dispersal in heterogeneous porous media

Microorganisms regulate the expression of energetically expensive phenotypes via a collective decision-making mechanism known as quorum sensing (QS). This study investigates the intricate dynamics of biofilm growth and QS-controlled biofilm dispersal in heterogeneous porous media, employing a pore-scale reactive transport modeling approach. Model simulations carried out under various fluid flow conditions and biofilm growth scenarios reveal that QS processes are influenced not only by the biomass density of biofilm colonies but also by a complex interplay between pore architecture, flow velocity, and the rates of biofilm growth and dispersal. This study demonstrates that pore architecture controls the initiation of QS processes and advection gives rise to oscillatory growth of biofilms. Such oscillation is suppressed if biofilm dynamics are in favor of sustaining a sufficiently high signal concentration, such as fast growth or slow dispersal rates. By establishing a mathematical framework, this study contributes to the fundamental understanding of QS-controlled biofilm dynamics in complex environments.

A novel perfusion bioreactor promotes the expansion of pluripotent stem cells in a 3D-bioprinted tissue chamber

While the field of tissue engineering has progressed rapidly with the advent of 3D bioprinting and human induced pluripotent stem cells (hiPSCs), impact is limited by a lack of functional, thick tissues. One way around this limitation is to 3D bioprint tissues laden with hiPSCs. In this way, the iPSCs can proliferate to populate the thick tissue mass prior to parenchymal cell specification. Here we design a perfusion bioreactor for an hiPSC-laden, 3D-bioprinted chamber with the goal of proliferating the hiPSCs throughout the structure prior to differentiation to generate a thick tissue model. The bioreactor, fabricated with digital light projection, was optimized to perfuse the interior of the hydrogel chamber without leaks and to provide fluid flow around the exterior as well, maximizing nutrient delivery throughout the chamber wall. After 7 days of culture, we found that intermittent perfusion (15 s every 15 min) at 3 ml min−1 provides a 1.9-fold increase in the density of stem cell colonies in the engineered tissue relative to analogous chambers cultured under static conditions. We also observed a more uniform distribution of colonies within the tissue wall of perfused structures relative to static controls, reflecting a homogeneous distribution of nutrients from the culture media. hiPSCs remained pluripotent and proliferative with application of fluid flow, which generated wall shear stresses averaging ∼1.0 dyn cm−2. Overall, these promising outcomes following perfusion of a stem cell-laden hydrogel support the production of multiple tissue types with improved thickness, and therefore increased function and utility.

First Report of Pestalotiopsis trachicarpicola Causing Apothecium Wither on Cultivated Morchella sextelata in Sichuan, China.

Morel (Morchella sextelata) is a valuable medicinal and edible mushroom, but the wild yield is seriously insufficient, so several artificial varieties have been developed to alleviate the shortage of wild yield. However, in 2020-2022, apothecium wither symptoms were observed in Nanchong, Sichuan. A total of 30.80% of the morels in the planting base (1.3 km2) showed these symptoms. The initial symptoms were slight white lesions on the surface of apothecium, then the mycelia gradually invaded the interior of the fruiting body, and eventually leading to black and wilt apothecium (Fig. 1a). Fifteen symptomatic morels were collected and ten isolates were obtained using the single spore isolation technique developed by Chomnunti et al. (2014). The morphological characteristics of the ten isolates were similar, which showed dense aerial white mycelia colony texture on PDA, and later forming concentric black mucus (Fig. 1c). The reverse side was yellow (Fig. 1d). The acervulus was floral-shaped and discrete, with smooth walls and measured 120.0 to 400.5 × 15.5 to 40.0 μm (n=10) (Fig. 1e); the conidia were fusiform and hyaline, 21.0 to 28.6 × 6.0 to 7.6 μm in size (n=50), each contained five cells. The apical cell was hyaline, conic and with 2 to 4 tubular apical appendages on the top. The three intermediate cells were brown to olivaceous, doliform to cylindrical, constricted at septa. The basal cell was conic to acute (Fig. 1f). The morphological characteristics were consistent with the published description of Pestalotiopsis trachicarpicola (Maharachchikumbura et al., 2012). PCR was performed with primers ITS1/ITS4 for internal transcribed spacer (ITS) (White et al., 1990), BT2A/BT2B for β-tubulin gene (TUB) (Glass and Donaldson, 1995), and EF1-526F/EF1-1567R for translation elongation factor 1-alpha (TEF-1α) (Roger et al., 1999). The pairwise alignments of ITS, TUB, and TEF-1α sequences was nearly 100% identical to P. trachicarpicola with GenBank accession numbers MT889666.1 (579/585 bp, 99%), MT884145.1 (445/450 bp, 99%), and MW149930.1 (946/958 bp, 99%), respectively. The resulting sequences were deposited in GenBank (Accession no. ITS: OL362082; TUB: OL828342; and TEF-1α: OL905009). Phylogenetic analysis performed with maximum likelihood method used MEGA 7.0 (1000 bootstrap replications) classified WLM5 into the P. trachicarpicola clade (Fig. 2), so we finally confirmed the identity of WLM5 as P. trachicarpicola. To fulfill Koch's postulates, twenty morels were surface disinfected with 2% sodium hypochlorite and then artificially wounded (diameter of 0.5 mm) prior to inoculation with 200 μL conidial suspension (105 conidia/mL), while an equal amount of sterile distilled water was applied to controls. After 4 days, the inoculated fruiting bodies showed symptoms consistent with field infection (Fig. 1b) and P. trachicarpicola was re-isolated using the same protocol, while the control remained asymptomatic. This first report of P. trachicarpicola causing apothecium wither on morel will help develop robust disease management strategies against this emerging fungal pathogen.

Natural recovery of corals after severe disturbance.

Ecosystem recovery from human-induced disturbances, whether through natural processes or restoration, is occurring worldwide. Yet, recovery dynamics, and their implications for broader ecosystem management, remain unclear. We explored recovery dynamics using coral reefs as a case study. We tracked the fate of 809 individual coral recruits that settled after a severe bleaching event at Lizard Island, Great Barrier Reef. Recruited Acropora corals, first detected in 2020, grew to coral cover levels that were equivalent to global average coral cover within just 2 years. Furthermore, we found that just 11.5 Acropora recruits per square meter were sufficient to reach this cover within 2 years. However, wave exposure, growth form and colony density had a marked effect on recovery rates. Our results underscore the importance of considering natural recovery in management and restoration and highlight how lessons learnt from reef recovery can inform our understanding of recovery dynamics in high-diversity climate-disturbed ecosystems.

Assessing the Relationship between Sea Turtle Strandings and Anthropogenic Impacts in Taiwan

Data acquired from stranded sea turtles can provide awareness of human activities that adversely affect sea turtle populations. We assessed strandings of five sea turtle species between 2017 and 2021. This study utilizes principal component analysis (PCA) and structural equation modeling (SEM) to reveal potential causes of sea turtle strandings linked to anthropogenic effects in Taiwan. Although our study did not observe a statistically significant impact of offshore wind turbines on sea turtle strandings, it did find evidence of a significant direct effect of coral colony density, heavy metals, and fishing disturbance on such strandings. For the conservation of endangered sea turtles, we recommend the incorporation of PCA and SEM in further contexts for validating anthropogenic impact assessments.

Density of wild honey bee, Apis mellifera, colonies worldwide.

The western honey bee, Apis mellifera, lives worldwide in approximately 102 million managed hives but also wild throughout much of its native and introduced range. Despite the global importance of A. mellifera as a crop pollinator, wild colonies have received comparatively little attention in the scientific literature and basic information regarding their density and abundance is scattered. Here, we review 40 studies that have quantified wild colony density directly (n = 33) or indirectly using genetic markers (n = 7) and analyse data from 41 locations worldwide to identify factors that influence wild colony density. We also compare the density of wild and managed colonies at a regional scale using data on managed colonies from the Food and Agriculture Organization (FAO). Wild colony densities varied from 0.1 to 24.2/km2 and were significantly lower in Europe (average of 0.26/km2) than in Northern America (1.4/km2), Oceania (4.4/km2), Latin America (6.7/km2) and Africa (6.8/km2). Regional differences were not significant after controlling for both temperature and survey area, suggesting that cooler climates and larger survey areas may be responsible for the low densities reported in Europe. Managed colony densities were 2.2/km2 in Asia, 1.2/km2 in Europe, 0.2/km2, in Northern America, 0.2/km2 in Oceania, 0.5/km2 in Latin America and 1/km2 in Africa. Wild colony densities exceeded those of managed colonies in all regions except Europe and Asia. Overall, there were estimated to be between two and three times as many wild colonies as managed worldwide. More wild colony surveys, particularly in Asia and South America, are needed to assess the relative density of wild and managed colonies at smaller spatial scales.

Reef structure of the Florida Reef Tract for the period 2005–2020

Shallow-water coral reefs of the Florida Reef Tract compose the third largest reef in the world, but during the last several decades, scleractinian (stony) corals have suffered unprecedented declines from global and local stressors. A program to evaluate the effects of high-temperature bleaching events was initiated by The Nature Conservancy’s Florida Reef Resilience Program in 2005 and surveys have been completed across at least some portion of the entire region every year since. The program adopted a demographic (colony-based) assessment approach, which records colony species, size (height and maximum diameter), and estimated partial mortality (percent barren skeleton). Because reef structure is critical to ecosystem functioning and services, data from 2005 to 2020 were analyzed to describe the abundance, size, and morphological complexity of stony coral colonies forming the biogenic reef. Colony height, footprint, surface area, and volume summed for 6016 transects were used to describe reef structure and averages were used to characterize the components that contributed to the structure. Nearly 150,000 colonies representing 49 species were reported during this period and results demonstrated both spatial and temporal changes for the region and for geographic subregions. Some subregions showed increasing colony density, especially for three small, hemispheric species, and declining average colony size.

Ecological and environmental predictors of juvenile coral density across the central and western Pacific

The persistence of Pacific coral reefs is threatened by large-scale mortality resulting from heat stress, storms, overfishing, disease, and land-based pollution. While reefs have demonstrated the ability to recover from these disturbances, recovery potential is highly variable; in part driven by the high spatial variability in the abundance of coral juveniles. However, our understanding of the patterns and predictors of juvenile coral density is hampered by the limited geographic scope of previous studies. Our objectives in this study were to: (1) explore the spatial and temporal patterns of juvenile (1- &amp;lt; 5cm in diameter) colony density across the U.S.-affiliated Pacific islands and atolls; and (2) identify the potential role of ecological and environmental factors which may influence coral juvenile abundance. Juvenile density was quantified as part of NOAA’s National Coral Reef Monitoring Program between 2013 and 2019 using a stratified sampling design across 1,405 forereef sites on 34 islands and atolls. Regional mean juvenile density varied from 1.4 to 10.5 colonies m-2 with the highest densities observed in the northern Mariana Islands, Northwestern Hawaiian Islands, and southern Mariana Islands. Juvenile density significantly increased between sampling periods in the Main Hawaiian Islands, Line Islands and northern Mariana Islands. Survey-weighted generalized linear modeling demonstrated that juvenile density was best predicted by the interaction between time since a heat stress event and heat stress severity, depth, site and sector-level percent coral cover, human density, percent unconsolidated substrate, macroalgal cover, herbivore biomass, and the interaction of sector coral cover area and heat stress, and the interaction of wave power (combination of wave height and period) and heat stress. With the unique scale and gradients across the U.S. Pacific, our study found that the high latitude regions have enhanced juvenile density compared to their lower latitude counterparts. Our results also suggest density is most dependent on the timing and severity of heat stress, depth, stock-recruitment with an open adult population, availability of suitable habitat at the site-scale, and the number of humans in the system.

Mine or my neighbours’ offspring: an experimental study on parental discrimination of offspring in a colonial seabird, the little auk Allealle

Individual recognition (IR) abilities may result from various ecological and naturally selected features of a species. Complex IR mechanisms should develop when the risk of misidentification of a chick is high. For colonial seabirds, the ability to identify their own brood is crucial to ensure parental fitness. Vocalizations seem to be a key component of most parent–offspring interactions, although few studies have assessed the interindividual differences in seabird chick calls and their potential usage in IR. The little auk (Allealle), which breeds in dense colonies, constitutes a perfect model for testing IR. In this study, we (1) examined chick calls at different stages of the nesting period, and (2) cross-fostered chicks to examine the rate of acceptance/nonacceptance of chicks by parents. We found significant interindividual differences in chick begging and fledging calls. Surprisingly, all cross-fostered chicks in our experiments were accepted by their foster parents, and male parents were as equally likely to accept cross-fostered chicks as females, even though the sexes would be expected to differ in offspring recognition due to different postfledging interactions with the chick. The revealed individuality of chick calls suggests the potential for chick vocal recognition in the studied species, but parent birds may disregard the individual characteristics enabling chick discrimination. This may take place as long as the chick is found in the nest because of the high likelihood that the chick present there is the focal one. However, IR during and after fledging requires further study. Studying the complexity of IR mechanisms is important for better understanding various avian social relationships and interactions.

The cotton mealybug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is spreading in North Africa: first report in Tunisia

ABSTRACT Biological invasions by alien agricultural insect pests represent nowadays a major threat to the durability of agricultural systems. In this study, we report for the first time the presence of an invasive economic pest of ornamental and solanaceous crops in Tunisia: the cotton mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae). It is native to America and has been spreading in Asia and the Mediterranean Basin. It was found in autumn 2021 forming dense colonies on Lantana camara L. (Verbenaceae) used for ornamental purposes in urban parks in the governorate of Tunis and on tomato Lycopersicon esculentum Mill. (Solanaceae) protected crops in the governorate of Sousse. Phenacoccus solenopsis was identified using an integrative approach based on morphological features and molecular analysis. Phylogenetic analysis using the mitochondrial Cytochrome Oxidase I (mtCOI) sequences of P. solenopsis populations from native and invaded areas retrieved in GenBank showed that Tunisian specimens might derive from an Asian population. We found that Nephus quadrimaculatus Herbst (Coleoptera: Coccinellidae: Scymninae) is the only natural enemy associated with this newly introduced pest so far. This is the first report of P. solenopsis attacking Lantana and tomato plants in Tunisia, and it should be considered as a potential threat to associated crops.

Woven bone formation and mineralization by rat mesenchymal stromal cells imply increased expression of the intermediate filament desmin.

Disordered and hypomineralized woven bone formation by dysfunctional mesenchymal stromal cells (MSCs) characterize delayed fracture healing and endocrine -metabolic bone disorders like fibrous dysplasia and Paget disease of bone. To shed light on molecular players in osteoblast differentiation, woven bone formation, and mineralization by MSCs we looked at the intermediate filament desmin (DES) during the skeletogenic commitment of rat bone marrow MSCs (rBMSCs), where its bone-related action remains elusive. Monolayer cultures of immunophenotypically- and morphologically - characterized, adult male rBMSCs showed co-localization of desmin (DES) with vimentin, F-actin, and runx2 in all cell morphotypes, each contributing to sparse and dense colonies. Proteomic analysis of these cells revealed a topologically-relevant interactome, focused on cytoskeletal and related enzymes//chaperone/signalling molecules linking DES to runx2 and alkaline phosphatase (ALP). Osteogenic differentiation led to mineralized woven bone nodules confined to dense colonies, significantly smaller and more circular with respect to controls. It significantly increased also colony-forming efficiency and the number of DES-immunoreactive dense colonies, and immunostaining of co-localized DES/runx-2 and DES/ALP. These data confirmed pre-osteoblastic and osteoblastic differentiation, woven bone formation, and mineralization, supporting DES as a player in the molecular pathway leading to the osteogenic fate of rBMSCs. Immunocytochemical and morphometric studies coupled with proteomic and bioinformatic analysis support the concept that DES may act as an upstream signal for the skeletogenic commitment of rBMSCs. Thus, we suggest that altered metabolism of osteoblasts, woven bone, and mineralization by dysfunctional BMSCs might early be revealed by changes in DES expression//levels. Non-union fractures and endocrine - metabolic bone disorders like fibrous dysplasia and Paget disease of bone might take advantage of this molecular evidence for their early diagnosis and follow-up.