Swarming Activity Research Articles

Complete genome sequence provides information on quorum sensing related spoilage and virulence of Aeromonas salmonicida GMT3 isolated from spoiled sturgeon

Foodborne bacteria can pose a threat to the public health due to their spoilage and virulence potential, which can be regulated by quorum sensing (QS) system. In the study, we isolated a spoilage bacteria strain Aeromonas salmonicida GMT3 from refrigerated sturgeon. The complete genome of A. salmonicida GMT3 was sequenced, and the QS related genes were assigned. QS signal molecules N-acyl-homoserine lactones (AHLs) and AI-2 were detected. Genes regulating the spoilage-related metabolic pathways, including protease and lipase secretion, amines metabolism, sulfur metabolism, motility and biofilm formation were analyzed. Furthermore, genes encoding for several virulence factors, e.g. hemolysin, aerolysin, type II secretion system (T2SS), type VI secretion system (T6SS), antibiotic and multidrug resistance were also identified. In addition, the spoilage and virulence phenotypes associated with QS including protease, swimming and swarming activity, biofilm and hemolytic activity were detected. This study provided new insights into spoilage and virulence mechanisms correlated with QS of A. salmonicida GMT3, which might promote development of new approaches for spoilage and virulence control based on QS target.

Roles of transcriptional factor PsrA in the regulation of quorum sensing in Pseudomonas aeruginosa PAO1.

The transcription factor PsrA regulates fatty acid metabolism, the type III secretion system, and quinolone signaling quorum sensing system in Pseudomonas aeruginosa. To explore additional roles of PsrA in P. aeruginosa, this study engineered a P. aeruginosa PAO1 strain to carry a recombinant plasmid with the psrA gene (pMMBpsrA) and examined the impact of elevated psrA expression to the bacterium. Transcriptomic analysis revealed that PsrA significantly downregulated genes encoding the master quorum-sensing regulators, RhlR and LasR, and influenced many quorum-sensing-associated genes. The role of PsrA in quorum sensing was further corroborated by testing autoinducer synthesis in PAO1 [pMMBpsrA] using two reporter bacteria strains Chromobacterium violaceum CV026 and Escherichia coli [pSB1075], which respond to short- and long-chain acyl homoserine lactones, respectively. Phenotypic comparisons of isogenic ΔpsrA, ΔlasR, and ΔpsrAΔlasR mutants revealed that the reduced elastase, caseinase, and swarming activity in PAO1 [pMMBpsrA] were likely mediated through LasR. Additionally, electrophoretic mobility shift assays demonstrated that recombinant PsrA could bind to the lasR promoter at a 5'-AAACGTTTGCTT-3' sequence, which displays moderate similarity to the previously reported consensus PsrA binding motif. Furthermore, the PsrA effector molecule oleic acid inhibited PsrA binding to the lasR promoter and restored several quorum sensing-related phenotypes to wild-type levels. These findings suggest that PsrA regulates certain quorum-sensing phenotypes by negatively regulating lasR expression, with oleic acid acting as a crucial signaling molecule.

Myrtus communis leaf compounds as novel inhibitors of quorum sensing-regulated virulence factors and biofilm formation: In vitro and in silico investigations

Antibiotic resistance of the Gram-negative bacterium Pseudomonas aeruginosa and its ability to form biofilm through the Quorum Sensing (QS) mechanism are important challenges in the control of infections caused by this pathogen. The extract of Myrtus communis (myrtle) showed strong anti-QS effect on Chromobacterium. violaceum 6267 by inhibiting 80 % of the production of violacein pigment at a sub-MIC concentration of 1/8 (31.25 μg/mL). In addition, the extract exhibited an inhibitory effect on virulence factors of P. aeruginosa PAO1 at half MIC (125 μg/mL), significantly reducing the formation of biofilms (72.02 %), the swarming activity (75 %), and the production of protease (61.83 %) and pyocyanin (97 %). The active fraction also downregulated the expression of selected regulatory genes involved in the biofilm formation and QS in the P. aeruginosa PAO1 strain. These genes included the autoinducer synthase genes (lasI and rhlI), the genes involved in the expression of their corresponding receptors (lasR and rhlR), and the pqsA genes. The analysis of the active fraction by HPLC/UV/MS and NMR allowed the identification of three phenolic compounds, 3,5-di-O-galloylquinic acid, myricetin 3-O-α-l-rhamnopyranoside (myricitrin), and myricetin 3-O-(2″-O-galloyl)-ß-d-galactopyranoside. In silico studies showed that 3,5-di-O-galloylquinic acid, with an affinity score of −9.20 kcal/mol, had the highest affinity to the active site of the CviR protein (3QP8), a QS receptor from C. violaceum. Additionally, myricetin 3-O-α-l-rhamnopyranoside (myricitrin) and myricetin 3-O-(2″-O-galloyl)-ß-d-galactopyranoside interact to a lesser extent with 3QP8. In conclusion, this study contributed significantly to the discovery of new QS inhibitors from M. communis leaves against resistant Gram-negative pathogens.

Enhancing Botrytis disease management in tomato plants: insights from a Pseudomonas putida strain with biocontrol activity.

This study explores the biocontrol potential of Pseudomonas putida Z13 against Botrytis cinerea in tomato plants, addressing challenges posed by the pathogen's fungicide resistance. The aims of the study were to investigate the in vitro and in silico biocontrol traits of Z13, identify its plant-colonizing efficacy, evaluate the efficacy of different application strategies against B. cinerea in planta, and assess the capacity of Z13 to trigger induced systemic resistance (ISR) in plants. The in vitro experiments revealed that Z13 inhibits the growth of B. cinerea, produces siderophores, and exhibits swimming and swarming activity. Additionally, the Z13 genome harbors genes that encode compounds triggering ISR, such as pyoverdine and pyrroloquinoline quinone. The in planta experiments demonstrated Z13's efficacy in effectively colonizing the rhizosphere and leaves of tomato plants. Therefore, three application strategies of Z13 were evaluated against B. cinerea: root drenching, foliar spray, and the combination of root drenching and foliar spray. It was demonstrated that the most effective treatment of Z13 against B. cinerea was the combination of root drenching and foliar spray. Transcriptomic analysis showed that Z13 upregulates the expression of the plant defense-related genes PR1 and PIN2 upon B. cinerea inoculation. The results of the study demonstrated that Z13 possesses significant biocontrol traits, such as the production of siderophores, resulting in significant plant protection against B. cinerea when applied as a single treatment to the rhizosphere or in combination with leaf spraying. Additionally, it was shown that Z13 root colonization primes plant defenses against the pathogen.

The behavior of the Bacillus probiotic species under conditions of co-cultivation

Combining probiotic bacteria is a promising strategy to increase the effectiveness and avoid side effects of probiotic therapy. Bacteria that find themselves in a common environment are able to both change their properties and show new ones under the influence of each other. The change of colonization and antagonistic properties, which provide bacteria with competitive advantages in the development of new spaces, deserves special attention.
 The aim of this research was to study the features of the mutual influence of probiotic bacilli: B. clausii, B. coagulans and B. subtilis on growth, swimming and swarming motility when co-cultivated on agar media of different solidity.
 Materials and methods. The study used commercial strains of bacilli from three probiotic preparations: Enterogermina, Lactovit forte and Subalin. The ability of the studied species of bacilli to influence each other’s growth was investigated by the agar block method (using 1.5 % nutrient agar) and the spot-on-lawn assay (using 0.7 % nutrient agar). The study of the mutual influence on swimming and swarming activity was carried out when bacilli were cultivated on 0.25 % and 0.70 % tryptone agar, respectively. The diameters of swimming halos and migration swarms formed by monoand mixed cultures were measured, and the areas covered by them were compared. The phenotypes of the meeting of swarms formed by cultures spotted on the swarm plates at different locations were also investigated.
 Results. The studied probiotic species of the bacilli did not show a strong ability to inhibit each other’s growth. The B. clausii culture had no inhibitory effect, and the B. coagulans culture demonstrated a moderate inhibitory influence on the growth of the other two species of bacilli when using both diffusion methods. The B. subtilis culture showed moderate or weak inhibitory activity against the B. clausii culture and weak or no inhibitory activity against the B. coagulans culture using the agar block method or spot-on-lawn assay, respectively. The B. coagulans + B. subtilis, B. clausii + B. coagulans and B. clausii + B. subtilis mixed cultures showed moderate, weak and no inhibitory activity against third cultures, respectively. The studied species of bacilli showed different swimming ability and swarming potential as well as the ability to influence each other’s motility. Swimming halos formed by the B. clausii + B. subtilis and B. clausii + B. сoagulans + B. subtilis mixed cultures covered significantly larger plate areas than the swimming halos formed by each culture separately during the same cultivation time. The highest swarming potential was observed in B. coagulans culture and B. cla usii + B. coagulans + B. subtilis mixed culture. The studied bacilli did not show the ability to merge swarms, but, on the contrary, their swarms at the point of contact formed visible “boundary” or “intermediate” lines, demonstrating the ability to identify nonself.
 Conclusions. The obtained results indicate the ability of the probiotic species of bacilli: B. clausii, B. coagulans and B. subtilis to mutually influence growth, swimming and swarm motility. The highest indicators of swimming and swarming of the triple mixed culture indicate an increase in the colonization potential of the studied bacilli when they are combined. The hypothesis of mutual induction of inhibitory compounds production by bacilli, which may increase the overall antagonistic potential of the triple mixed culture, is subject to further confirmation.

Comprehensive genomic analysis of Bacillus paralicheniformis strain BP9, pan-genomic and genetic basis of biocontrol mechanism

Many Bacillus species are essential antibacterial agents, but their antibiosis potential still needs to be elucidated to its full extent. Here, we isolated a soil bacterium, BP9, which has significant antibiosis activity against fungal and bacterial pathogens. BP9 improved the growth of wheat seedlings via active colonization and demonstrated effective biofilm and swarming activity. BP9 sequenced genome contains 4282 genes with a mean G-C content of 45.94% of the whole genome. A single copy concatenated 802 core genes of 28 genomes, and their calculated average nucleotide identity (ANI) discriminated the strain BP9 from Bacillus licheniformis and classified it as Bacillus paralicheniformis. Furthermore, a comparative pan-genome analysis of 40 B. paralicheniformis strains suggested that the genetic repertoire of BP9 belongs to open-type genome species. A comparative analysis of a pan-genome dataset using the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Cluster of Orthologous Gene groups (COG) revealed the diversity of secondary metabolic pathways, where BP9 distinguishes itself by exhibiting a greater prevalence of loci associated with the metabolism and transportation of organic and inorganic substances, carbohydrate and amino acid for effective inhabitation in diverse environments. The primary secondary metabolites and their genes involved in synthesizing bacillibactin, fencing, bacitracin, and lantibiotics were identified as acquired through a recent Horizontal gene transfer (HGT) event, which contributes to a significant part of the strain`s antimicrobial potential. Finally, we report some genes essential for plant-host interaction identified in BP9, which reduce spore germination and virulence of multiple fungal and bacterial species. The effective colonization, diverse predicted metabolic pathways and secondary metabolites (antibiotics) suggest testing the suitability of strain BP9 as a potential bio-preparation in agricultural fields.

Similarities in social calls during autumn swarming may facilitate interspecific communication between Myotis bat species

Bats employ a variety of social calls for communication purposes. However, for most species, social calls are far less studied than echolocation calls and their specific function often remains unclear. We investigated the function of in-flight social calls during autumn swarming in front of a large hibernaculum in Northern Germany, whose main inhabitants are two species ofMyotisbats, Natterer’s bats (Myotis nattereri) and Daubenton’s bats (Myotis daubentonii). We recorded social calls in nights of high swarming activity and grouped the calls based on their spectro-temporal structure into ten types and verified our visual classification by a discriminant function analysis. Whenever possible, we subsequently assigned social calls to eitherM. daubentoniiorM. nattereriby analyzing the echolocation calls surrounding them. As many bats echolocate at the same time during swarming, we did not analyze single echolocation calls but the “soundscape” surrounding each social call instead, encompassing not only spectral parameters but also the timbre (vocal “color”) of echolocation calls. Both species employ comparatively similar social call types in a swarming context, even though there are subtle differences in call parameters between species. To additionally gain information about the general function of social calls produced in a swarming context, we performed playback experiments with free-flying bats in the vicinity of the roost, using three different call types from both species, respectively. In three out of six treatments, bat activity (approximated as echolocation call rate) increased during and after stimulus presentation, indicating that bats inspected or approached the playback site. Using a camera trap, we were sometimes able to identify the species of approaching bats. Based on the photos taken during playbacks, we assume one call type to support interspecific communication while another call type works for intraspecific group cohesion.

Versatile role of Pseudomonas fuscovaginae cyclic lipopeptides in plant and microbial interactions.

Pseudomonas fuscovaginae is the most prominent bacterial sheath rot pathogen, causing sheath brown rot disease in rice. This disease occurs worldwide and it is characterized by typical necrotic lesions on the sheath, as well as a reduction in the number of emitted panicles and filled grains. P. fuscovaginae has been shown to produce syringotoxin and fuscopeptin cyclic lipopeptides (CLPs), which have been linked to pathogenicity. In this study, we investigated the role of P. fuscovaginae UPB0736 CLPs in plant pathogenicity, antifungal activity and swarming motility. To do so, we sequenced the strain to obtain a single-contig genome and we constructed deletion mutants in the biosynthetic gene clusters responsible for the synthesis of CLPs. We show that UPB0736 produces a third CLP of 13 amino acids, now named asplenin, and we link this CLP with the swarming activity of the strain. We could then show that syringotoxin is particularly active against Rhizoctonia solani in vitro. By testing the mutants in planta we investigated the role of both fuscopeptin and syringotoxin in causing sheath rot lesions. We proved that the presence of these two CLPs considerably affected the number of emitted panicles, although their number was still significantly affected in the mutants deficient in both fuscopeptin and syringotoxin. These results reveal the importance of CLPs in P. fuscovaginae pathogenicity, but also suggest that other pathogenicity factors may be involved.

The soundscape of swarming: Proof of concept for a noninvasive acoustic species identification of swarming Myotis bats.

Bats emit echolocation calls to orientate in their predominantly dark environment. Recording of species-specific calls can facilitate species identification, especially when mist netting is not feasible. However, some taxa, such as Myotis bats can be hard to distinguish acoustically. In crowded situations where calls of many individuals overlap, the subtle differences between species are additionally attenuated. Here, we sought to noninvasively study the phenology of Myotis bats during autumn swarming at a prominent hibernaculum. To do so, we recorded sequences of overlapping echolocation calls (N= 564) during nights of high swarming activity and extracted spectral parameters (peak frequency, start frequency, spectral centroid) and linear frequency cepstral coefficients (LFCCs), which additionally encompass the timbre (vocal "color") of calls. We used this parameter combination in a stepwise discriminant function analysis (DFA) to classify the call sequences to species level. A set of previously identified call sequences of single flying Myotis daubentonii and Myotis nattereri, the most common species at our study site, functioned as a training set for the DFA. 90.2% of the call sequences could be assigned to either M. daubentonii or M. nattereri, indicating the predominantly swarming species at the time of recording. We verified our results by correctly classifying the second set of previously identified call sequences with an accuracy of 100%. In addition, our acoustic species classification corresponds well to the existing knowledge on swarming phenology at the hibernaculum. Moreover, we successfully classified call sequences from a different hibernaculum to species level and verified our classification results by capturing swarming bats while we recorded them. Our findings provide a proof of concept for a new noninvasive acoustic monitoring technique that analyses "swarming soundscapes" by combining classical acoustic parameters and LFCCs, instead of analyzing single calls. Our approach for species identification is especially beneficial in situations with multiple calling individuals, such as autumn swarming.

Silicone Foley catheters impregnated with microbial indole derivatives inhibit crystalline biofilm formation by Proteus mirabilis.

Proteus mirabilis is a common causative agent for catheter-associated urinary tract infections (CAUTI). The crystalline biofilm formation by P. mirabilis causes catheter encrustation and blockage leading to antibiotic treatment resistance. Thus, biofilm formation inhibition on catheters becomes a promising alternative for conventional antimicrobial-based treatment that is associated with rapid resistance development. Our previous work has demonstrated the in vitro antibiofilm activity of microbial indole derivatives against clinical isolates of P. mirabilis. Accordingly, we aim to evaluate the capacity of silicone Foley catheters (SFC) impregnated with these indole derivatives to resist biofilm formation by P. mirabilis both phenotypically and on the gene expression level. Silicon Foley catheter was impregnated with indole extract recovered from the supernatant of the rhizobacterium Enterobacter sp. Zch127 and the antibiofilm activity was determined against P. mirabilis (ATCC 12435) and clinical isolate P8 cultured in artificial urine. The indole extract at sub-minimum inhibitory concentration (sub-MIC=0.5X MIC) caused a reduction in biofilm formation as exhibited by a 60-70% reduction in biomass and three log10 in adhered bacteria. Results were confirmed by visualization by scanning electron microscope. Moreover, changes in the relative gene expression of the virulence genes confirmed the antibiofilm activity of the indole extract against P. mirabilis. Differential gene expression analysis showed that extract Zch127 at its sub-MIC concentration significantly down-regulated genes associated with swarming activity: umoC, flhC, flhD, flhDC, and mrpA (p< 0.001). In addition, Zch127 extract significantly down-regulated genes associated with polyamine synthesis: speB and glnA (p< 0.001), as well as the luxS gene associated with quorum sensing. Regulatory genes for capsular polysaccharide formation; rcsB and rcsD were not significantly affected by the presence of the indole derivatives. Furthermore, the impregnated catheters and the indole extract showed minimal or no cytotoxic effect against human fibroblast cell lines indicating the safety of this intervention. Thus, the indole-impregnated catheter is proposed to act as a suitable and safe strategy for reducing P. mirabilis CAUTIs.

Demographic characteristics shape patterns of dawn swarming during roost switching in tree-dwelling Daubenton’s bat

Frequent roost switching in fission–fusion societies of tree-dwelling bats is closely associated with swarming behaviour entailing ritualised night-time displays around the roost tree and/or at the roost entrance to signal its actual location, particularly immediately prior to sunrise. However, effects of demographic characteristics of individuals in this social behaviour remain unanswered. Using passive integrated transponders (PIT) and automatic readers, we recorded swarming activity of members of a Daubenton’s bat (Myotis daubentonii) maternity colony in the vicinity of their roosts. In total, 59,622 activity events of 281 PIT-tagged individuals were recorded on ten monitored roosts during three summer seasons. We found a gradual increase of swarming activity from midnight to sunrise in old adult females, whereas young females and juveniles primarily swarmed later at dawn. We attribute this difference to the learning status of younger bats, which are not yet able to perform a defined pattern of swarming activity, whereas older bats likely take a more active role in signalling the position of the roost. Old males exhibited the least swarming activity at maternity roosts, which mostly occurred between crepuscular periods, presumably due to their solitary lives. A negative correlation between genetic distance and swarming activity suggests an important role of kinship in the formation of the maternity colony as well as group cohesion during roost switching.

Stopping the swarm

Immunology Neutrophils play a major role in the early immune response and are recruited in large numbers into inflamed and infected tissues. By secreting chemoattractants that bind G protein–coupled receptors (GPCRs) on neighboring cells, neutrophils coordinate their behavior as a swarm. Less clear is how this auto-amplifying swarming activity is ultimately turned off. Kienle et al. show that desensitization of these GPCRs by the same chemoattractants by GPCR-kinase 2 (GRK2) is one way in which these swarms are shut down (see the Perspective by Rocha-Gregg and Huttenlocher). Unexpectedly, mice with GRK2-deficient neutrophils showed impaired rather than enhanced bacterial clearance. The heightened scanning ability of GRK2-deficient neutrophils may come at the cost of suboptimal phagocytosis and containment of bacteria. Science , abe7729, this issue p. [eabe7729][1]; see also abj3065, p. [1262][2] [1]: /lookup/doi/10.1126/science.abe7729 [2]: /lookup/doi/10.1126/science.abj3065

Neutrophils self-limit swarming to contain bacterial growth in vivo.

Neutrophils communicate with each other to form swarms in infected organs. Coordination of this population response is critical for the elimination of bacteria and fungi. Using transgenic mice, we found that neutrophils have evolved an intrinsic mechanism to self-limit swarming and avoid uncontrolled aggregation during inflammation. G protein-coupled receptor (GPCR) desensitization acts as a negative feedback control to stop migration of neutrophils when they sense high concentrations of self-secreted attractants that initially amplify swarming. Interference with this process allows neutrophils to scan larger tissue areas for microbes. Unexpectedly, this does not benefit bacterial clearance as containment of proliferating bacteria by neutrophil clusters becomes impeded. Our data reveal how autosignaling stops self-organized swarming behavior and how the finely tuned balance of neutrophil chemotaxis and arrest counteracts bacterial escape.

Adult males in maternity colonies of Daubenton’s bat, Myotis daubentonii: what are they?

Abstract During summer, females of temperate zone bats form maternity colonies consisting almost exclusively of mothers and their offspring. However, in some species the proportion of males in nurseries is relatively high. We explored the presence of these males in Daubenton’s bat, with respect to their age and relatedness to the local maternity colony. Using automatic PIT-tag readers, we recorded individual swarming activity at roosts used for day roosting and discovered that most of the males were yearlings that returned to their nursery roost after hibernation. The elevated occurrence of yearling males in spring could be explained by late mating attempts or toleration in the territory of adults.

Exploring the quorum sensing inhibition of isolated chrysin from Penicillium chrysogenum DXY-1

We recently and for the first time reported that ethyl acetate extracts isolated from Penicillium chrysogenum DXY-1 exhibited anti-quorum sensing (anti-QS) activity. Herein, another active molecule in the extracts was identified as chrysin by NMR and MS. A 20 μg/mL dose of chrysin inhibited violacein production regulated by QS in C. violaceum CV026 by 31.6%. A 40 μg/mL dose of chrysin suppressed pyocyanin production, elastase activity, proteolytic activity, and biofilm formation regulated by QS in P. aeruginosa PA01 by 41.4%, 13.8%, 8.3%, and 42.4%, respectively. And chrysin could inhibit the swarming activity of P. aeruginosa PA01. Further, molecular docking and CD analysis were used to address the mechanism of chrysin's activity in C. violaceum. Molecular docking results revealed that chrysin suppresses QS system by competing with the natural signal molecule C6HSL for binding to the same pocket of CviR receptor. At the same time, CD results also showed that chrysin could change the secondary structure composition of CviR, which greatly prevented the binding of C6HSL/CviR, and further playing its role on inhibiting bacterial QS system. All these data demonstate that chrysin may be used as a potential QS inhibitor to tackle increasing drug resistance.

Entomological Surveillance and Cantharidin Concentrations in Mylabris variabilis and Epicauta rufidorsum Blister Beetles in Slovenia.

Simple SummaryTrue blister beetles (genus Epicauta, family Meloidae) produce cantharidin, which is a potent blistering agent. Cantharidin toxicosis is well documented in humans and animals. Drought and modern harvesting techniques are thought to increase the likelihood of blister beetle contamination of forage and cantharidin intoxication in animals. Local presence, behavioral patterns, and cantharidin concentration were studied in Mylabris variabilis and Epicauta rufidorsum. In this study, E. rufidorsum was found to be the most likely source of forage contamination due to its local abundance, swarming activity, and tendency to reside in the green parts of plants after cutting. Cantharidin was detected in the bodies of both blister beetles species studied. It is likely that modern forage harvesting methods, which involve simultaneous processing of forage after cutting, increase the incidence of cantharidin toxicoses in animals. Delaying these processes by a few minutes would reduce the likelihood of Epicauta contamination of the forage.True blister beetles (genus Epicauta, family Meloidae) produce cantharidin, which can cause toxicosis in humans and animals. Some recent reports suggest that poisoning by the blister beetle has occurred in the Mediterranean part of Slovenia, which has never been reported before. Drought and modern harvesting techniques are thought to increase the likelihood of blister beetle forage contamination and cantharidin intoxication in animals. A survey of fields associated with blister beetle contamination was conducted and the Meloid species present were identified. Entomological surveillance was conducted for Mylabris variabilis and Epicauta rufidorsum. Cantharidin concentrations were also measured in both blister beetle species. Cantharidin concentration in Mylabris variabilis (n = 17) ranged from 0.038 to 0.354 µg/mg (mean 0.151 µg/mg). Cantharidin concentration in Epicauta rufidorsum (n = 36) ranged from 0.055 to 0.341 µg/mg (mean 0.142 µg/mg). Both species exhibited variable concentrations of cantharidin that could not be associated with their biology, sex, age, size, and/or reproductive status. Epicauta rufidorsum have never previously been studied as a possible source of forage contamination, nor have cantharidin concentrations been determined in this species. It is the most likely source of forage contamination due to its abundance in the investigated fields, its swarming activity, and its tendency to reside in the green parts of plants immediately after cutting. Delaying the simultaneous processing and storage of forage after cutting would reduce the likelihood of forage contamination by blister beetles, as they can then retreat to the ground or fly away.

Virulence potential of Aeromonas hydrophila isolated from apparently healthy freshwater food fish

Aeromonas hydrophila is considered an important pathogen of fish causing substantial economic loss in aquaculture worldwide. The objective of the current study was to assess the virulence potential of A. hydrophila strains isolated from gills of apparently healthy fish using phenotypic assays, PCR based genotypic tests, and in-vivo pathogenicity assays in Indian catfish, Clarias magur. For this, motile aeromonads (n = 50) were isolated from gills of apparently healthy freshwater food fish (n = 50) collected from domestic fish market. Gill swabs were incubated for primary enrichment in alkaline peptone water supplemented with cephalothin (10 mg/mL) for 12 hours followed by plating on Starch Ampicillin Agar. A single big yellow colony was selected from each sample, purified, and presumptively identified as Aeromonas with the help of Aerokey-II. Out of the fifty isolates of Aeromonas, five isolates of diverse origin (9C, 10G1, 7C, 10C and 4P) were confirmed to be A. hydrophila by automated bacterial identification system VITEK 2 with confidence interval of 90–98%. In the phenotypic assays, strains 9C and 10G1 showed high serum resistance, swimming and swarming activity and low biofilm producing capabilities, which were indicative of putative virulence. Both the strains belonged to genotype aah+act+ alt+ascV+eno+lip+ast+ on the basis of detection of virulence genes by PCR, i.e. extracellular haemolysin (aah), cytolytic enterotoxin (act), heat-labile cytotonic enterotoxin (alt), type-III secretion system (ascV), enolase (eno), lipase (lip) and heat-stable cytotonic enterotoxin (ast). In-vivo pathogenicity assays confirmed that both the strains were pathogenic to magur fish. The LD50 for 9C strain was 6.81 × 104 CFU/mL and 7.62 × 105 CFU/mL for 10G1strain. In conclusion, our phenotypic and genotypic findings showed that A. hydrophila isolated from apparently healthy fish harbour number of important virulence genes/factors and could have important implications in triggering disease in farmed fish under stress.

An agent-based algorithm resembles behaviour of tree-dwelling bats under fission\u2013fusion dynamics

Utilization of computational approach in the study of social behaviour of animals is increasing and we attempted such an approach in our study of tree-dwelling bats. These bats live in highly dynamic fission–fusion societies that share multiple roosts in a common home range. The key behavioural component associated with complex and non-centralized decision-making processes in roost switching is swarming around potential locations in order to recruit members to the new roost. To understand roost switching dynamics of bat groups in their natural environment, we employed a computational model, the SkyBat, which is based on swarm algorithm, to model this process. In a simulated environment of this agent-based model, we replicated natural fission–fusion dynamics of the Leisler’s bat, Nyctalus leisleri, groups according to predefined species and habitat parameters. Spatiotemporal patterns of swarming activity of agents were similar to bats. The number of simulated groups formed prior to sunrise, the mean number of individuals in groups and the roost height did not differ significantly from data on a local population of bats collected in the field. Thus, the swarm algorithm gave a basic framework of roost-switching, suggesting possible applications in the study of bat behaviour in rapidly changing environments as well as in the field of computer science.

Validation of Horizontal Genes Transfer Using Selected Clinical Strain of P. Mirabsilis for Swarming Activity

Introduction: Proteus mirabilis expresses several virulence factor which enable them to colonize, surviveand grow in the host organism. This study was to investigate whether P. mirabilis could deliver a spontaneousplasmids to competent strains, and to investigate whether P. mirabilis can be as a donor strains to deliverytheir virulent such as adhesion and swarming activity.Methodology: PCR was used to detect the motility genes that encoded in P. mirabilis. Using alsotransformation assay to transfer these genes of Proteus to standard strain. PCR technique used again todetermine the whether genes were transfer or not after transformation.The results: the findings show that 9 (filD, flhC, flhD, motA, motB, fliL, flaA, flgM,) genes were encodedon chromosomal DNA of P. mirabilis. Interestingly data, only 3 (flaA, flgM and filD ) genes were carriedon different plasmids of P. mirabilis. The results also show that spontaneous plasmids of P. mirabilis cantransfer to standard strains of E. coli. We found one gene transfer to E. coli S17? filD. The gene(filD) has arole in the P. mirabilis swarming motility.Conclusions: we demonstrated that these genes can carry not only on chromosomal DNA but also inPlasmidal DNA and transferring genes associated-characteristics, swarming motility

Tracing the geographic origin of common pipistrelles (Pipistrellus pipistrellus) swarming at a mass hibernaculum

Mass swarming of tens of thousands of common pipistrelles in front of hibernacula of the Carpathian Mountains suggests that bats may originate from a large catchment area. However, until now neither banding nor molecular data have resolved the geographic origin of common pipistrelles at these sites. Here, we measured the acoustic activity of bats and the stable hydrogen isotope ratios (δ2H) in metabolically inert fur keratin of bats to infer the relative swarming activity and the putative summer origin of bats, respectively, observed in autumn at Erňa cave, one of the largest bat hibernacula in Europe. Swarming activity declined with decreasing ambient temperature during the early season, while it increased during colder days towards the onset of hibernation. Based on δ2H values, we deduced that about 50% of the animals did not have a local origin. Provenance of all but one of these migrants was identified as the Pannonian Basin, while a single long-distance migrant may have originated from the northern margin of the species’ European distribution range. Modelling the variation in δ2H values of bats in response to sex, body condition and season suggested that, towards the onset of hibernation, males of low body condition were likely to be of distant geographic origin. Throughout the swarming season, females were mostly of local origin, yet towards the onset of hibernation, their body condition was not as variable as in males. We conclude that common pipistrelles observed at this mass hibernacula site are facultative migrants, which may undertake long-distance seasonal movements occasionally. At our study site, common pipistrelles are more likely to include long-distance migrants because of the mass occurrence of this species at this large hibernaculum.