Phylogenetic Groups Research Articles

Prevalence and molecular characterization of ESBL-producing Escherichia coli isolated from broiler chicken and their respective farms environment in Malaysia.

Extended spectrum beta-lactamase-producing Escherichia coli (ESBL-EC) is an increasing public health threat. This study aimed to determine the prevalence and characterization of ESBL-producing Escherichia coli (E. coli) isolated from broiler chicken and their farm environment, in Kelantan Malaysia. Escherichia coliwas isolated from 453 collected samples, including 210 cloacal swabs and 243 environmental samples. The antimicrobial susceptibility profile of the E. coli isolates was assessed for sixteen antibiotics using the disc diffusion method. The E. coli isolates were evaluated for phenotypic ESBL production using modified double disc synergy. After extraction of genomic DNA, ESBL resistance genes, phylogenetic group, and virulence genes were detected by PCR using appropriate primers. ESBL genes were further confirmed by sequencing. The molecular typing of E. coli strains was determined by Multilocus Sequence Typing (MLST). A total of 93.8% (425/453) E. coli were isolated from the collected samples. Out of 334 E. coli isolates screened, 14.7% (49/334) were phenotypically ESBL producers. All the ESBL-EC were resistant to tetracycline, ciprofloxacin, and ampicillin. Thus, 100% of the ESBL-EC were multidrug resistant. Of the ESBL-EC 81.6% were positive for at least one ESBL encoding gene. The most prevalent ESBL gene detected was blaTEM (77.6%; 38/49) followed by blaCTX-M (32.7%; 16/49) and blaSHV (18.4%; 9/49). The majority of ESBL-EC belonged to phylogenic groups A followed by B1 accounting for 44.9% and 12.2%, respectively. The most frequently identified sequence types were ST10 (n = 3) and ST206 (n = 3). The most detected virulence genes in the E. coli isolates were astA (33.3%; 22/66) followed by iss (15.2%; 10/66). Our results show both broiler chicken and their respective farms environment were reservoirs of multi-drug resistant ESBL-producing E. coli and ESBL resistance genes.

Evolution of copper resistance in Xanthomonas euvesicatoria pv. perforans population.

The widespread use of antimicrobials that target bacterial pathogens has driven evolution of resistance, compromising the efficacy of these bactericides. Understanding the emergence and spread of resistance genes via mobile genetic elements is crucial for combating antimicrobial resistance. Copper resistance (CuR) in Xanthomonas euvesicatoria pv. perforans has severely affected the efficacy of copper-based bactericides for controlling bacterial leaf spot disease of tomato and pepper. Here, we investigated the evolutionary pathways of CuR acquisition and dissemination in X. euvesicatoria pv. perforans using an extensive collection of strains. We determined that chromosomally encoded CuR predominates over plasmid-borne CuR in multiple distinct phylogenetic groups of X. euvesicatoria pv. perforans. Our analysis revealed a single site of chromosomal integration by a CuR genomic island, although the genomic island showed sequence variation among phylogenetic groups. While chromosomal CuR was more prevalent, strains with plasmid-borne resistance conferred greater copper tolerance. Additionally, we identified strains carrying two copies of CuR genes, on plasmid and chromosome, that exhibited increased copper tolerance. Strains of X. euvesicatoria pv. perforans from the USA shared identical CuR gene sequences whether on plasmids or chromosome while different alleles were found in strains from other countries. In contrast to X. euvesicatoria pv. perforans, plasmid-borne CuR predominated in closely related pathovar, X. euvesicatoria pv. euvesicatoria. Overall, these findings contribute to a better understanding of the evolution and persistence of CuR in X. euvesicatoria pv. perforans and its closest relatives.IMPORTANCEThe emergence of antimicrobial resistance is a significant threat to agricultural production as it reduces the efficacy of various antimicrobials including copper-based bactericides that are widely used to control plant diseases. The challenge of increasing antimicrobial resistance entering a production system necessitates a deeper understanding of the dynamics and mechanisms by which pathogens acquire resistance. As a result of this research, we have identified different mechanisms of copper resistance acquisition as well as levels of copper resistance in a devastating plant pathogen, X. euvesicatoria pv. perforans. The evolution and dissemination of copper resistance in strains through plasmid or chromosomally integrated genomic island or both presents barriers to current management approaches, where growers rely heavily on copper-based bactericides to manage disease outbreaks. This knowledge is crucial when considering the continued use of existing antimicrobials or adopting alternative antimicrobials in efforts to implement enhanced antimicrobial stewardship strategies in agriculture.

Individual, ontogenetic, and phylogenetic variation in the dentition of hadrosaurids (Iguanodontia: Ornithischia)

Phylogenetic analyses of Hadrosauroidea are generally well-resolved, but finer resolution within Hadrosauromorpha remains contentious. This lack of resolution is due in part to the inability of discrete traits to illustrate their natural variation, particularly relating to the dental tooth battery. Not only has the variation not been properly examined, but the character definitions fluctuate throughout the literature. Here, we evaluate individual, ontogenetic and phylogenetic variation in four fundamental tooth characters at the subfamily level to better establish their use in future analyses: number of tooth rows, tooth aspect ratio, secondary enamel ridges and marginal enamel denticles. Factor analysis of mixed data and linear discriminant analysis were used to evaluate how accurately these four characters together identified the phylogenetic groups. We then used these data to predict phylogenetic relationships for a select few historically problematic taxa. We found that although the number of tooth rows is phylogenetically informative, it is related to size and is only useful in separating hadrosaurids from non-hadrosaurid iguanodontians. Tooth aspect ratio was found to be highly variable, and phylogenetic groups cannot be separated reliably from one another. Secondary ridges are individually variable, and their presence or absence should be grouped into a single character instead of being separated. Lastly, the presence and shape of marginal denticles in hadrosaurids is similarly variable, though rounded mammillations are almost exclusive to Lambeosaurinae, with some variation in their presence and prevalence within the group. When used in combination, these characters can accurately identify dentary teeth below Hadrosauridae, providing new options for identifying poorly preserved specimens and isolated teeth from microsites. Taken together, these characters may still be informative, but the individual variation described here must be accounted for when constructing character states, and we recommend several modifications to their coding.

Impact of the Seasons on the Evolution of Tomato-infecting Geminivirus and Identification of Plant Reservoirs Harboring Geminivirus in Burkina Faso

Geminiviruses are known as responsible for huge damage to vegetable crops in many tropical, subtropical, and temperate regions. In Burkina Faso, tomato was reported as the most infected vegetable crop with the involvement of several geminivirus species despite its socioeconomic importance. This suggests and underlines the pressing need for additional information on virus population evolution (diversity, prevalence, and host plant reservoirs). Thus, to address this problem, surveys and sample collection were carried out in three localities in Burkina Faso (Goué, Léguéma, and Toussiana). Tomato leaves and those of cultivated and non-cultivated plants around the tomato fields were collected. The total DNA extracted from these samples was subjected to diagnosis based on the polymerase chain reaction (PCR) protocol with or without amplicon sequencing. Results showed that depending on the season and locality, the prevalence of the disease varied from 0 to 53.33% and that of the virus species varied from 0 to 76.67%. The highest prevalence was observed in the dry season. In addition, PepYVMLV was the virus that was widely detected in all three localities in the rainy (18.89%) and dry (52.22%) seasons. The DNA-B molecule associated with this virus was associated with other begomoviruses (ToLCMLV). In addition, out of 200 samples collected in the vicinity of the tomato fields, only 63 were positive based on PCR diagnosis. Amplicon sequencing yielded 63 partial sequences of virus from ten plant species, including six non-cultivated species. Based on phylogenetic analysis, the 63 partial sequences belonged to three phylogenetic groups (the ToLCV group, the PepYVMLV group, and the CLCuGV group). This study allowed a better understanding of the evolution of tomato leaf curl/yellow disease in Burkina Faso and the diversity of plant species serving as reservoirs for involved viruses. This constitutes an important step in the search for adequate control methods.

What constitutes a stigma? A review of isolated pores in raphid diatoms (Bacillariophyceae) and the value of precise terminology

AbstractScanning electron microscopy has revealed variation in the ultrastructure of distinctive isolated pores in or near the central area of raphid diatoms, with different types of pores being restricted to phylogenetic groups. Thus, the widespread use of the term stigma for all such pores not only hides the structural diversity but also obscures the phylogenetic distribution of the different types. This paper provides images of the different types of isolated pores, particularly refining the discrimination of variants within the Cymbellales, and reveals some interesting ecological patterns. Revised definitions of stigmata and stigmoids are proposed, together with the recognition and definition of another type of stigmoid. The restricted distribution of more precisely defined pore types shows the importance of consistent use of terminology and its relevance to phylogenetic studies.

Interrelation Between Pathoadaptability Factors and Crispr-Element Patterns in the Genomes of Escherichia coli Isolates Collected from Healthy Puerperant Women in Ural Region, Russia.

Escherichia coli is a commensal and opportunistic bacterium widely distributed around the world in different niches including intestinal of humans and animals, and its extraordinary genome plasticity led to the emergence of pathogenic strains causing a wide range of diseases. E. coli is one of the monitored species in maternity hospitals, being the main etiological agent of urogenital infections, endometriosis, puerperal sepsis, and neonatal diseases. This study presents a comprehensive analysis of E. coli isolates obtained from the maternal birth canal of healthy puerperant women 3-4 days after labor. According to whole genome sequencing data, 31 sequence types and six phylogenetic groups characterized the collection containing 53 isolates. The majority of the isolates belonged to the B2 phylogroup. The data also includes phenotypic and genotypic antibiotic resistance profiles, virulence factors, and plasmid replicons. Phenotypic and genotypic antibiotic resistance testing did not demonstrate extensive drug resistance traits except for two multidrug-resistant E. coli isolates. The pathogenic factors revealed in silico were assessed with respect to CRISPR-element patterns. Multiparametric and correlation analyses were conducted to study the interrelation of different pathoadaptability factors, including antimicrobial resistance and virulence genomic determinants carried by the isolates under investigation. The data presented will serve as a valuable addition to further scientific investigations in the field of bacterial pathoadaptability, especially in studying the role of CRISPR/Cas systems in the E. coli genome plasticity and evolution.

Extended-spectrum beta-lactamase-producing Enterobacterales in human health: Experience from the tricycle project, Ghana.

Vulnerable groups, such as pregnant women, are at increased risk of potentially life-threatening infections with extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-E) for both mother and newborn. However, data regarding ESBL-E carriage and associated risk factors in Ghanaian pregnant women remain scarce. This study aimed to determine the prevalence of ESBL-E carriage and its associated risk factors among pregnant women attending the antenatal clinic at the Korle Bu Teaching Hospital. A systematic sample of 700 pregnant women with gestational age ≥ 34 weeks attending the antenatal clinic at Korle Bu Teaching Hospital was included in the study. After administering a structured questionnaire to assess potential risk factors associated with ESBL-E carriage, patients were given a sterile stool container to submit at least 1 g of stool specimen. Recovered isolates from faecal specimens were identified using MALDI-TOF-MS technology. These isolates were then subjected to susceptibility testing and ESBL identification. A random subset of 24 ESBL-producing Escherichia coli isolates was whole-genome sequenced on the MiSeq Illumina platform. Risk factors associated with ESBL-E carriage were determined using multivariable logistic regression analysis. Among the 700 pregnant women, 42% (294) carried ESBL-E. The predominant ESBL-producing Enterobacterales were Escherichia coli (95%). Fifty percent (50%) of ESBL-E were multidrug resistant isolates (MDRs). Whole-genome sequencing of 24 ESBL-producing E. coli isolates revealed that blaCTX-M-15 (96%) was the most prevalent ESBL gene type. Notably, most isolates belonged to commensal phylogenetic groups (A, B1, and C; 88%). Having a primary level of education (aOR 1.45, 95% CI 1.05-1.96) and consuming legumes as the main source of protein (aOR 0.17, 0.40-0.83) were significantly associated with intestinal carriage of ESBL-E. This study identified a high prevalence of ESBL-E and MDR-ESBL-E carriage among pregnant women. Our findings underscore the urgent need for public health interventions to control the spread of AMR.

Genome-Wide Identification of Basic Helix-Loop-Helix (bHLH) Family in Peanut: Potential Regulatory Roles in Iron Homeostasis.

The basic helix-loop-helix (bHLH) superfamily is the second-largest transcription factor family that participates in a wide range of biological processes in plants, including iron homeostasis. Although the family has been studied in several plant species, a comprehensive investigation is still needed for peanut (Arachis hypogaea). Here, a genome-wide analysis identified 373 AhbHLH genes in peanut, which were divided into 14 groups or subfamilies according to phylogenetic analysis. Clustered members generally share similar gene/protein structures, supporting the evolutionary relationships among AhbHLH proteins. Most AhbHLHs experienced whole-genome or segmental duplication. The majority of AhbHLH proteins had a typical bHLH domain, while several phylogenetic groups, including Group VI, X, XIII, and XIV, had the HLH domain. The expression of several AhbHLH genes, including AhbHLH001.3, AhbHLH029.1/.2, AhbHLH047.1/.2, AhbHLH115.1/.2, AhbHLH097.1/.2, AhbHLH109.4, and AhbHLH135.1, was induced by Fe deficiency for both cultivars, or at least in Silihong, suggesting an important role in the Fe deficiency response in peanut. Nine genes (AhbHLH001.3, AhbHLH029.1/.2, AhbHLH047.1/.2, AhbHLH097.1/.2, and AhbHLH115.1/.2) were specifically induced by Fe deficiency in Silihong, and their expression was higher in Silihong than that in Fenghua 1. These genes might be responsible for higher tolerance to Fe deficiency in Silihong. Our findings provide comprehensive information for further elucidating the regulatory mechanism of Fe homeostasis in peanut.

Understanding the anatomical diversity of leaves in the Aspicarpoid clade: New insights for Malpighiaceae systematics

The Aspicarpoid clade represents a strongly supported group within the Stigmaphylloid lineage. The group is composed of four neotropical genera: Aspicarpa Rich., Gaudichaudia Kunth, Janusia A.Juss. and Peregrina W.R.Anderson. In this study, we studied the utility of leaf anatomy characters to support groups recovered in molecular phylogenies and define synapomorphies that can be used in future infra- and supra-generic classifications. Camarea axillaris A.St.-Hil. (1823: 10) and Cottsia Dubard & Dop (1908: 20) were used as outgroups for comparison. Dry leaf samples were re-hydrated, sectioned, stained, and analyzed using a microscope and a stereomicroscope. Characters such as presence and absence of glands, their location and morphotype, petiole and midrib contour, presence of trichomes, accessory bundles, vascular system organization and mesophyll type were utilized for species comparison, showing high taxonomic value for the group. Leaf glands, accessory bundles, sheath extension in the mesophyll and presence of druses and prismatic crystals were the most valuable characters in an evolutionary approach for the clade. Our work confirms the power of leaf anatomy as a taxonomic and evolutionary tool in Malpighiaceae Juss. (1789: 252), as well as its contribution to the delimitation of phylogenetic groups in the Aspicarpoid clade.

Characterization of virus species associated with sweet potato virus disease in Costa Rica and riboprobe development for its rapid detection

AbstractSweet potato (Ipomoea batatas) is a crucial crop for food security and economic stability in many regions, including Costa Rica. This study aimed to assess the prevalence and genetic diversity of two major viruses associated with sweet potato virus disease (SPVD) in Costa Rica: sweet potato feathery mottle virus (SPFMV) and sweet potato chlorotic stunt virus (SPCSV). A total of 50 leaf samples displaying typical viral disease symptoms were collected from major sweet potato‐growing regions. Using multiplex reverse transcription (RT)‐PCR, SPFMV was detected in 82% of the samples and SPCSV in 52%, with a coinfection rate of 48%. Genetic analysis revealed that SPFMV isolates belonged predominantly to phylogenetic group B, while SPCSV isolates clustered with the West African (WA) strain. The low genetic diversity within these viral populations suggests a limited number of initial infection events followed by local spread. Riboprobe technology was evaluated as an alternative to RT‐PCR for virus detection. While the riboprobes for SPFMV demonstrated sensitivity comparable to RT‐PCR, the SPCSV riboprobes showed slightly lower sensitivity, particularly when using crude leaf extracts. Despite this, riboprobes offer significant advantages in terms of cost and ease of use for large‐scale screening. Our findings underscore the need for robust diagnostic and management strategies to control SPVD and highlight the importance of continuous monitoring and research to mitigate the impact of viral diseases on sweet potato production in Costa Rica. These insights contribute to the global understanding of SPVD and support collaborative efforts in plant pathology research.

Listeria monocytogenes in Fruits and Vegetables: Antimicrobial Resistance, Biofilm, and Genomic Insights.

Listeria monocytogenes is a foodborne pathogen that can infect both humans and animals and cause noninvasive gastrointestinal listeriosis or invasive listeriosis. The objectives of this study were to determine the genetic diversity of L. monocytogenes; the genes associated with its resistance to antibiotics, benzalkonium chloride (BC), and cadmium chloride (CdCl2); and its biofilm formation. A total of 132 fresh fruits (44 samples) and vegetables (88 samples) were selected for this study. The genetic diversity of the isolates and the genes associated with their antibiotic resistance were determined using PCR amplification; meanwhile, their levels of susceptibility to antibiotics were determined using the agar diffusion method. Their levels of resistance to BC and CdCl2 were determined using the minimum inhibitory concentration method, and their capacity for biofilm formation was evaluated using the crystal violet staining method. A total of 17 L. monocytogenes strains were collected: 12.8% (17/132) from fresh fruits and vegetables in this study. The isolates of L. monocytogenes belonged to phylogenetic groups I.1 (29.4% (5/17); serotype 1/2a) and II.2 (70.5% (12/17); serotype 1/2b); strains containing Listeria pathogenicity islands (LIPIs) were also identified at prevalence rates of 100% for LIPI-1 and LIPI-2 (17/17), 29.4% for LIPI-3 (5/17), and 11.7% for LIPI-4 (2/17). The antibiotic susceptibility tests showed that the L. monocytogenes isolates exhibited six different multiresistant patterns, with multiple antibiotic resistance (MAR) index of ≥0.46 (70.5%; 12/17); additionally, the genes Ide, tetM, and msrA, associated with efflux pump Lde, tetracycline, and ciprofloxacin resistance, were detected at 52.9% (9/17), 29.4% (5/17), and 17.6% (3/17), respectively. The phenotypic tests showed that 58.8% (10/17) of cadmium-resistant L. monocytogenes isolates had a co-resistance of 23.5% (4/17) to BC. Finally, all strains of L. monocytogenes exhibited moderate biofilm production. The results of this study contribute to our understanding of the persistence and genetic diversity of L. monocytogenes strains isolated from fresh fruits and vegetables; in addition, their resistance to CdCl2, which is correlated with co-resistance to BC disinfectant, is helpful for the food industry.

Identifying in vitro toxicity testing approaches for (novel) proteins in the context of food and feed risk assessment

Abstract This report documents the outcomes of the EFSA procurement (OC/EFSA/NIF/2022/01) aimed at identifying in vitro toxicity testing approaches for (novel) proteins in the context of food and feed safety assessment. In the present report, we present an integrated testing strategy for the evaluation of toxicity of novel/toxic proteins. A text‐mining approach was used to create a literature database of toxic outcomes associated with toxic proteins retrieved from the UniProt KB database using the search term “Toxin activity”. It was shown that toxic proteins are produced by a relatively limited phylogenetic subset, including, among others, bacteria, insects, serpents, molluscs, and fungi. Toxicological effects of these proteins are generally conserved within phylogenetic groups. Analysis of toxic effects from these proteins was performed using GO term analysis as well as a text‐mining based approach. Relevant tests to address and quantify these toxicity effects were identified and evaluated for their applicability in an in vitro based toxicity testing strategy. A stepwise approach was developed. As a first step, an initial in silico prediction of toxicity is carried out (Step 1). This is followed by a battery of in vitro assays to address the primary mechanisms of toxicity associated with toxic proteins (Step 2). If concern arises in the Step 2 battery of tests, the use of relevant in vitro model systems to explore potential target organ toxicity are required (Step 3). Knowledge gaps have been identified and recommendations are provided in in vitro toxicity testing strategies, in particular for (novel) proteins. Some of these gaps involve the selection and integration of a standardized, relevant in vitro digestion step, reflective of passage through the digestive tract, within the testing strategy, as well as a thorough assessment of the suitability and applicability of in vitro tests and new approach methodologies for regulatory toxicity assessment of (novel) proteins. To accelerate the incorporation of NAMs in the assessment of protein safety, case studies and proof of concept projects are needed to demonstrate the utility and effectiveness of in vitro toxicity testing strategies in the safety assessment of (novel) proteins.

Comparative genomic analysis and characterization of novel high-quality draft genomes from the coal metagenome.

Coal, a sedimentary rock harbours a complex microbial community that plays a significant role in its formation and characteristics. However, coal metagenome sequencing and studies were less, limiting our understanding of this complex ecosystem. This study aimed to reconstruct high-quality metagenome-assembled genomes (MAGs) from the coal sample collected in the Neyveli mine to explore the unrevealed diversity of the coal microbiome. Using Illumina sequencing, we obtained high-quality raw reads in FASTQ format. Subsequently, de novo assembly and binning with metaWRAP software facilitated the reconstruction of coal MAGs. Quality assessment using CheckM identified 10 High-Quality MAGs (HQ MAGs), 7 medium-quality MAGs (MQ MAGs), and 6 low-quality MAGs (LQ MAGs). Further analysis using GTDB-Tk revealed four HQ MAGs as known species like Dermacoccus abyssi, Sphingomonas aquatilis, Acinetobacter baumannii, and Burkholderia cenocepacia. The remaining six HQ MAGs were classified as Comamonas, Arthrobacter, Noviherbaspirillum, Acidovorax, Oxalicibacterium, and Bordetella and designated as novel genomes by the validation of digital DNA-DNA hybridization (dDDH). Phylogenetic analysis and further pangenome analysis across the phylogenetic groups revealed a similar pattern with a high proportion of cloud genes. We further analysed the functional potential of these MAGs and closely related genomes using COG. The comparative functional genomics revealed that novel genomes are highly versatile, potentially reflecting adaptations to the coal environment. BlastKOALA was used to conduct a detailed analysis of the metabolic pathways associated with the MAGs. This study highlights the comparative genomic analysis of novel coal genomes with their closely related genomes to understand the evolutionary relationships and functional properties.

Virulence Factors, Phylogenetic Groups and Ciprofloxacin Susceptibility of Escherichia coli Isolated from Healthy Students in Chungcheong Province

Virulence Factors, Phylogenetic Groups and Ciprofloxacin Susceptibility of Escherichia coli Isolated from Healthy Students in Chungcheong Province

Identification of AP2/ERF Transcription Factors and Characterization of AP2/ERF Genes Related to Low-Temperature Stress Response and Fruit Development in Luffa

Plant-specific APETALA2/Ethylene-Responsive Factor (AP2/ERF) transcription factors are involved in the regulation of genes associated with the growth and developmental processes of numerous plants. Although AP2/ERF proteins from other species have been intensively studied, no studies have been reported on the AP2/ERF family of Luffa cylindrica, an important vegetable of the cucurbit family, and one of the most popular vegetables in the world. In this study, 133 genes (315–6696 bp) encoding LcAP2/ERF proteins with complete AP2/ERF domains were identified according to the luffa P93075 genome. These LcAP2/ERF genes were subsequently classified and analyzed for their gene structures, chromosomal distribution locations, promoter cis-acting elements, conserved structural domains of encoded proteins, and responses to abiotic stresses. The LcAP2/ERF genes were identified and divided into five phylogenetic groups (AP2, DREBs, ERFs, RAV, and soloists). These genes were unevenly distributed across 13 chromosomes. An analysis of gene structures indicated the LcAP2/ERF genes contained 0–11 introns (average of 4.4). Additionally, 16 motifs were identified in the LcAP2/ERF proteins that were conserved across different phylogenetic groups. Moreover, 11 cis-acting elements associated with response to the environment were analyzed in a 2000 bp region upstream of the LcAP2/ERF gene promoters. A transcriptome analysis involving RNA-seq data revealed tissue-specific LcAP2/ERF expression profiles and the diversity in LcAP2/ERF expression. The effects of low-temperature stress on LcAP2/ERF expression were determined. Furthermore, fruit-development-related and low-temperature-induced expressional changes were verified by RT-qPCR analyses of 14 differentially expressed LcAP2/ERF genes in luffa. Our findings will help clarify the evolution of the luffa AP2/ERF family, while also providing valuable insights for future studies on AP2/ERF functions.

Ethylene and epoxyethane metabolism in methanotrophic bacteria: comparative genomics and physiological studies using Methylohalobius crimeensis.

The genome of the methanotrophic bacterium Methylohalobius crimeensis strain 10Ki contains a gene cluster that encodes a putative coenzyme-M (CoM)-dependent pathway for oxidation of epoxyethane, based on homology to genes in bacteria that grow on ethylene and propylene as sole substrates. An alkene monooxygenase was not detected in the M. crimeensis genome, so epoxyethane is likely produced from co-oxidation of ethylene by the methane monooxygenase enzyme. Similar gene clusters were detected in about 10% of available genomes from aerobic methanotrophic bacteria, primarily strains grown from rice paddies and other wetlands. The sparse occurrence of the gene cluster across distant phylogenetic groups suggests that multiple lateral gene transfer events have occurred in methanotrophs. In support of this, the gene cluster in M. crimeensis was detected within a large genomic island predicted using multiple methods. Growth studies, reverse transcription-quantitative PCR (RT-qPCR) and proteomics were performed to examine the expression of these genes in M. crimeensis. Growth and methane oxidation activity were completely inhibited by the addition of >0.5% (v/v) ethylene to the headspace of cultures, but at 0.125% and below, the inhibition was only partial, and ethylene was gradually oxidized. The etnE gene encoding epoxyalkane:CoM transferase was strongly upregulated in ethylene-exposed cells based on RT-qPCR. Proteomics analysis confirmed that EtnE and nine other proteins encoded in the same gene cluster became much more predominant after cells were exposed to ethylene. The results suggest that ethylene is strongly inhibitory to M. crimeensis, but the bacterium responds to ethylene exposure by expressing an epoxide oxidation system similar to that used by bacteria that grow on alkenes. In the obligate methanotroph M. crimeensis, this system does not facilitate growth on ethylene but likely alleviates toxicity of epoxyethane formed through ethylene co-oxidation by particulate methane monooxygenase. The presence of predicted epoxide detoxification systems in several other wetland methanotrophs suggests that co-oxidation of ambient ethylene presents a stress for methanotrophic bacteria in these environments and that epoxyethane removal has adaptive value.

Interactions between sexual signaling and wing size drive ecology and evolution of wing colors in Odonata

Insect coloration has evolved in response to multiple pressures, and in Odonata (dragonflies and damselflies) a body of work supports a role of wing color in a variety of visual signals and potentially in thermoregulation. Previous efforts have focused primarily on melanistic coloration even though wings are often multicolored, and there has yet to be comprehensive comparative analyses of wing color across broad geographic regions and phylogenetic groups. Percher vs. flier flight-style, a trait with thermoregulatory and signaling consequences, has not yet been studied with regard to color. We used a new color clustering approach to quantify color across a dataset of over 8,000 odonate wing images representing 343 Nearctic species. We then utilized phylogenetically informed Bayesian zero-inflated mixture models to test how color varies with mean ambient temperature, body size, sex and flight-style. We found that wing coloration clustered into two groups across all specimens - light brown-yellow and black-dark brown - with black-dark brown being a much more cohesive grouping. Male perchers have a greater proportion of black-dark brown color on their wings as do species with longer wings. In colder climates, odonates were more likely to have black-dark brown color present, but we found no relationship between the proportion of black and temperature. Light brown-yellow showed similar scaling with wing length, but no relationship with temperature. Our results suggest that black-dark brown coloration may have a limited role in thermoregulation, while light brown-yellow does not have such a role. We also find that the odonate sexes are divergent in wing color in percher species only, suggesting a strong role for color in signaling in more territorial males. Our research contributes to an understanding of complex interactions driving ecological and evolutionary dynamics of color in animals.

Longitudinal study of avian pathogenic Escherichia coli (APEC) serogroups associated with disease in Georgia poultry using molecular serology and virulence gene analysis

ABSTRACT Avian pathogenic Escherichia coli (APEC) is a significant cause of morbidity, mortality, and production loss to the poultry industry worldwide. Here, we characterized 569 E. coli isolates from avian-diagnosed colibacillosis cases from the state of Georgia, USA. In total, 339 isolates were assigned into 32 serogroups with the majority classifying as O78, O2, O25, O8, O1, O86, O18, and O15. Serogroup O25 was found to link with broilers, while broiler breeders were more often associated with serogroup O1 and pet/ hobby birds with serogroup O8. In addition, some serogroups (O1) were more prevalent in the summer and fall. Analysis for virulence-associated genes (VAGs) found 23.20% of isolates did not harbour any genes linked with the APEC pathotype, while ColV plasmid-associated genes (iroN, ompT, hlyF, iss, and aerJ) were frequently detected among most isolates (with 80–96% prevalence) and some of these genes were linked with serogroup. Phylogenetic analysis, classified isolates into phylogenetic groups B2 (34%), F (19%), A (15%), and G (9%). The phylogenetic group B2 isolates also harboured the highest number of VAGs. This study highlights that the current APEC-causing disease in birds in the State of Georgia has identified several emerging serogroups possessing several VAGs that could potentially lead to challenges in colibacillosis control. RESEARCH HIGHLIGHTS Several emerging APEC serogroups were observed in Georgia poultry populations. An association between APEC serogroups and bird type was observed. The prevalence of different APEC serogroups was influenced by season. A multiplex PCR assay targeting common serogroups of APEC in Georgia poultry was developed.

Sample Preparation for Proteogenomics of Aquatic Invertebrates.

Aquatic invertebrates account for many known animal species. Their diversity of form and lifestyle and adaptation to various habitats are outstanding. Several of these animals are considered reliable indicators of watershed health and are thus frequently used as environmental sentinels in ecotoxicology programs. Although aquatic invertebrates cover a large spectrum of the Tree of Life, few representatives have been sufficiently studied at the most intimate molecular level to be considered reference models. Proteogenomics has completely changed the game since the full molecular characterization of a proteome can be obtained straightforwardly for any invertebrate, thus helping to define conserved features or specific characteristics of distinct phylogenetic groups and to propose biomarkers for environmental monitoring. Typically, to obtain the most comprehensive analysis, a database of putative protein sequences is informed by sequencing full transcripts. The extracted proteins are proteolyzed, and the resulting peptides are resolved via reverse-phase liquid chromatography and identified by high-resolution tandem mass spectrometry. The data analysis makes it possible to establish a list of animal proteins and their abundances. Herein, we describe a sample preparation method for protein extraction from small aquatic invertebrates and for performing shotgun proteogenomics suitable for crustaceans and mollusks, which are the most common aquatic invertebrates.

Insights into Genes Encoding LEA_1 Domain-Containing Proteins in Cyperus esculentus, a Desiccation-Tolerant Tuber Plant

LEA_1 domain-containing proteins constitute a class of late-embryogenesis-abundant proteins that are highly hydrophilic and predominantly accumulate in mature seeds. Though LEA_1 proteins have been proven to be essential for seed desiccation tolerance and longevity, little information is available on their roles in non-seed storage organs. In this study, a first genome-wide characterization of the LEA_1 gene family was conducted in tigernut (Cyperus esculentus L., Cyperaceae), whose underground tubers are desiccation tolerant with a moisture content of less than 6%. Five family members identified in tigernut are comparative to four to six found in seven other Cyperaceae plants, but relatively more than three reported in Arabidopsis. Further comparison of 125 members from 29 plant species supports early divergence of the LEA_1 family into two phylogenetic groups before angiosperm radiation, and gene expansion in tigernut was contributed by whole-genome duplications occurring after the split with the eudicot clade. These two phylogenetic groups could be further divided into six orthogroups in the momocot clade, five of which are present in tigernut and the remaining one is Poaceae specific. Frequent structural variation and expression divergence of paralogs were also observed. Significantly, in contrast to seed-preferential expression of LEA_1 genes in Arabidopsis, rice, and maize, transcriptional profiling and qRT-PCR analysis revealed that CeLEA1 genes have evolved to predominantly express in tubers, exhibiting a seed desiccation-like accumulation during tuber development. Moreover, CeLEA1 transcripts in tubers were shown to be considerably more than that of their orthologs in purple nutsedge, another Cyperaceae plant producing desiccation-sensitive tubers. These results imply species-specific activation and key roles of CeLEA1 genes in the acquisition of desiccation tolerance of tigernut tubers as observed in orthodox seeds. Our findings not only improve the understanding of lineage-specific evolution of the LEA_1 family, but also provide valuable information for further functional analysis and genetic improvement in tigernut.