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Research on the initial phage-host interaction has been conducted on a limited repertoire of phages and their cognate receptors, such as phage λ and the Escherichia coli LamB (EcLamB) protein. Apart from phage λ, little is known about other phages that target EcLamB. Here, we developed a simple method for isolating novel environmental phages in a predictable way, i.e. isolating phages that target a particular receptor(s) of a bacterium, in this case, the EcLamB protein. A plasmid (pMUT13) encoding the EcLamB porin was transferred into three different enterobacterial genera. By enrichment with these engineered bacteria, a number of phages (ZZ phages) that targeted EcLamB were easily isolated from the environment. Interestingly, although EcLamB-dependent in their recombinant heterologous hosts, these newly isolated ZZ phages also targeted OmpC as an alternative receptor when infecting E. coli. Moreover, the phage host range was readily extended within three different bacterial genera with heterologously expressed EcLamB. Unlike phage λ, which is a member of the Siphoviridae family, these newly isolated EcLamB-dependent phages were more commonly members of the Myoviridae family, based on transmission electron microscopy and genomic sequences. Modifications of this convenient and efficient phage enrichment method could be useful for the discovery of novel phages., (© 2020 The Author. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

The phage life cycle is a multi-stage process initiated by the recognition and attachment of the virus to its bacterial host. This adsorption step depends on the specific interaction between bacterial structures acting as receptors and viral proteins called Receptor Binding Proteins (RBP). The adsorption process is essential as it is the first determinant of phage host range and a sine qua non condition for the subsequent conduct of the life cycle. In phages belonging to the Caudoviricetes class, the capsid is attached to a tail, which is the central player in the adsorption as it comprises the RBP and accessory proteins facilitating phage binding and cell wall penetration prior to genome injection. The nature of the viral proteins involved in host adhesion not only depends on the phage morphology (i.e., myovirus, siphovirus, or podovirus) but also the targeted host. Here, we give an overview of the adsorption process and compile the available information on the type of receptors that can be recognized and the viral proteins taking part in the process, with the primary focus on phages infecting Gram-positive bacteria. [ABSTRACT FROM AUTHOR]

Aeromonas veronii is a pathogen which can induce diseases in humans, animals and aquatic organisms, but its pathogenic mechanism and virulence factors are still elusive. In this study, we successfully constructed a mutant strain (ΔascP) by homologous recombination. The results showed that the deletion of the ascP gene significantly down-regulated the expression of associated effector proteins in A. veronii compared to its wild type. The adhesive and invasive abilities of ΔascP to EPC cells were 0.82-fold lower in contrast to the wild strain. The toxicity of ΔascP to cells was decreased by about 2.91-fold (1 h) and 1.74-fold (2 h). Furthermore, the LD50 of the mutant strain of crucian carp was reduced by 19.94-fold, and the virulence was considerably attenuated. In contrast to the wild strain, the ΔascP content in the liver and spleen was considerably lower. The titers of serum cytokines (IL-8, TNF-α, and IL-1β) in crucian carp after the infection of the ΔascP strain were considerably lower in contrast to the wild strain. Hence, the ascP gene is essential for the etiopathogenesis of A. veronii TH0426. [ABSTRACT FROM AUTHOR]

Dietary fibre substances in dog foods are composed of cell wall polysaccharides, non-cellulose polysaccharides, and structural non-polysaccharides. The purpose of this study was to determine the effects of dog age and heat-steam pressure (extruding process) on the in vitro fermentation of fibrous feedstuffs (sugar beet pulp, tomato pomace, wheat bran, corn bran, and rice bran) to be included in dog foods. The fibrous feedstuffs were incubated with a fermentation medium mixture and faeces inoculum, which were collected from two 6 month old (puppy), two 2 year old (adult), and two 8 year old (geriatric) dogs. The in vitro cumulative gas production (at 6, 12, 18, 24, 36, and 48 h), true-organic matter disappearance (T-OMd) (at 6, 12, 24, and 48 h), and molarities of short chain fatty acids (SCFA) (acetic, propionic, and butyric acids; AA, PA, and BA) in fermentation fluid (at 24 h) of fibrous feedstuffs were determined. The extruding process increased the in vitro cumulative gas production of rice bran (at 12, 18, 24, and 48 h) and wheat bran (at 6, 12, 24, and 36 h) (P < 0.05). In addition, in vitro cumulative gas production values of tomato pomace (at 12, 18, 24, 36, and 48 h) were reduced by the extruding process (P < 0.05). The extruding process increased the in vitro T-OMd values of sugar beet pulp (at 6, 12, and 24 h), wheat and rice bran (at 6 and 12 h), and the molarities of AA, BA, and total SCFA of the in vitro fermentation fluid of sugar beet pulp and wheat bran (P < 0.05). The in vitro cumulative gas production values of tomato pomace from faecal inoculums of the puppy and adult dogs were higher than that of the geriatric dog (P < 0.05). The in vitro T-OMd of sugar beet pulp (at 48 h) and extruded corn bran (at 6 h) with faecal inoculums of adult dog was higher than those of inoculum from puppy faeces (P < 0.05). The molarity of AA of tomato pomace with adult dog's faecal inoculum was higher than those from puppy and geriatric dog (P < 0.05). The molarities of AA, BA, and total SCFA with corn bran from faeces inoculums of the puppy and adult dog was higher than that of the geriatric dog (P < 0.05). The molarities of AA and total SCFA of the in vitro fermentation fluid of extruded rice bran with faecal inoculums from the geriatric dogs was higher than those with faecal inoculums from the puppy and adult dogs (P < 0.05). As a result, the extruding process positively affected the in vitro fermentation values of sugar beet pulp, wheat bran, and rice bran. Furthermore, the organic acid molarities of the in vitro fermentation fluid of extruded rice bran from the geriatric dog was higher than those from the puppy and adult dogs. Sugar beet pulp, tomato pomace, wheat bran, and corn bran can be used as a source of fibre in food for puppy, adult, and geriatric large dogs. [ABSTRACT FROM AUTHOR]

Vaccines represent one of the most promising strategies for mitigating economic losses imposed by infectious diseases to global aquaculture. While the majority of commercial vaccines employ injection as the main route of delivery, a substantial body of research explores other avenues for the stimulation of protective immunity. Oral vaccines, representing the most favourable and convenient solution for the aquaculture industry, have been at the forefront of scientific interest for the last couple of decades. The orchestrated effort resulted in identifying the main roadblocks on the path to an ideal edible vaccine and provided several ingenious solutions improving the activation of immunity, ensuring the stability of administered antigens and their uptake upon the passage through the stomach. In the presented review, we describe advances in the available knowledge of the processes prerequisite for developing protective mucosal vaccines and focus readers' attention on the untapped potential of plant‐based production systems in this effort. We propose that these approaches not only meet production demands but also fulfil all requirements of an oral vaccine, addressing all the obstacles, previously solved via separate strategies, in one platform. Thus, combined with the available knowledge of molecular adjuvants and produced for a fraction of the cost, plant‐based production, so‐called molecular farming, is an ideal candidate for vaccine development, paving the road to sustainable aquaculture. [ABSTRACT FROM AUTHOR]

This study evaluated the effect of a flushing diet containing docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) from lemuru fish oil in ewe ration on colostrum quality and lamb performance. Twenty Garut ewes were divided into four treatments of flushing concentrate: control concentrate (P1), flushing concentrate containing 6% palm oil (P2), flushing concentrate containing 3% lemuru oil and 3% palm oil (P3), flushing concentrate containing 6% lemuru oil (P4). The animal consumed Napier grass and concentrate with a diet ratio of 30%:70% based on dry matter. The experimental ewes were fed flushing rations two weeks before and two weeks after mating continued with 2 weeks before and two weeks after lambing. The ewes and their lambs were put together until weaning time with around two months. The parameters observed in ewes were nutrient consumption and their colostrum qualities. Meanwhile, the lamb parameters measured were nutrient consumption, lamb blood metabolites at birth, and lamb performance. The experimental design used a randomized blocked design (RBD). Analysis of variance (ANOVA) followed by Duncan test was used to analyze data. The results showed that treatment did not significantly affect dry matter consumption in ewe and lamb, ewe's colostrum quality, and lamb's blood metabolites. However, the treatment significantly improved (p<0.05) average daily gain and weaning weights of lambs. In conclusion, the flushing ration did not affect the quality of colostrum produced. Feeding of experimental ewes 2 weeks before and 2 weeks after mating continued with 2 weeks before and 2 weeks after parturition with flushing rations of 6% lemuru oil containing EPA and DHA can produce twin lambs with good growth performances such as daily gain and weaning weight similar to the control, which has single litter size. [ABSTRACT FROM AUTHOR]

Phages are regarded as powerful antagonists of bacteria, especially in industrial fermentation processes involving bacteria. While bacteria have developed various defense mechanisms, most of which are effective against a narrow range of phages and consequently exert limited protection from phage infection. Here, we report a strategy for developing phage-resistant Escherichia coli strains through the simultaneous genomic integration of a DNA phosphorothioation-based Ssp defense module and mutations of components essential for the phage life cycle. The engineered E. coli strains show strong resistance against diverse phages tested without affecting cell growth. Additionally, the resultant engineered phage-resistant strains maintain the capabilities of producing example recombinant proteins, D-amino acid oxidase and coronavirus-encoded nonstructural protein nsp8, even under high levels of phage cocktail challenge. The strategy reported here will be useful for developing engineered E. coli strains with improved phage resistance for various industrial fermentation processes for producing recombinant proteins and chemicals of interest. Phage contamination is a persistent problem in industrial biotechnology processes employing bacterial strains. Here, the authors report the construction of E. coli host strains with broad antiphase activities via the genomic integration of the Ssp defense system and mutations of components essential for phage infection cycles. [ABSTRACT FROM AUTHOR]

Linkages between the outer membrane of Gram-negative bacteria and the peptidoglycan layer are crucial for the maintenance of cellular integrity and enable survival in challenging environments1–5. The function of the outer membrane is dependent on outer membrane proteins (OMPs), which are inserted into the membrane by the β-barrel assembly machine6,7 (BAM). Growing Escherichia coli cells segregate old OMPs towards the poles by a process known as binary partitioning, the basis of which is unknown8. Here we demonstrate that peptidoglycan underpins the spatiotemporal organization of OMPs. Mature, tetrapeptide-rich peptidoglycan binds to BAM components and suppresses OMP foldase activity. Nascent peptidoglycan, which is enriched in pentapeptides and concentrated at septa9, associates with BAM poorly and has little effect on its activity, leading to preferential insertion of OMPs at division sites. The synchronization of OMP biogenesis with cell wall growth results in the binary partitioning of OMPs as cells divide. Our study reveals that Gram-negative bacteria coordinate the assembly of two major cell envelope layers by rendering OMP biogenesis responsive to peptidoglycan maturation, a potential vulnerability that could be exploited in future antibiotic design.Peptidoglycan stem peptides in the Gram-negative bacterial cell wall regulate the insertion of essential outer membrane proteins, thus representing a potential target for antibiotic design. [ABSTRACT FROM AUTHOR]

In this article I address three main crises of the Anthropocene: energy and climate, biodiversity and food, and peace and security. To obtain overall sustainability, natural scientists and engineers have developed the Energy Transition Model, the Carbon Transition Model, and the Agri-food-nature Transition Model. From these models two facts stand out. The age of fossil fuels and of livestock farming will come to an end. Because a sustainable society also has sociopolitical and legal, socioeconomic and financial, sociocultural and welfare, and socioenvironmental and ecological aspects, the contribution of human scientists to overall sustainability is also crucial. [ABSTRACT FROM AUTHOR]

Bacteriophage λ is a temperate virus infecting the bacterium Escherichia coli. Temperate phages have the ability to form lysogens where the prophage is maintained in the infected bacterium in a largely quiescent state. The lysogen becomes immune to superinfection by λ by blocking the development of the superinfecting phage. Here we report the λ lysogen not only protected itself from killing by λ phages but also protected λ-sensitive bacteria in mixed culture. This protection required that the lysogen was able to adsorb the superinfecting λ phages. The protection was also sensitive to the growth state of the mixed culture, and the λ lysogen lost efficiency in protecting λ-sensitive bacteria as it stopped growing. A mutant of the λ tail protein, λJ, was not subject to this loss of protection. Adsorption of λ having the wild-type J protein but not the mutant λJ protein to E. coli was inhibited by interference with bacterial energy metabolism. The last observation suggests wild-type λ preferentially infects bacteria with competent energy metabolism. [ABSTRACT FROM AUTHOR]

Viruses and their hosts can undergo coevolutionary arms races where hosts evolve increased resistance and viruses evolve counter‐resistance. Given these arms race dynamics (ARD), both players are predicted to evolve along a single trajectory as more recently evolved genotypes replace their predecessors. By coupling phenotypic and genomic analyses of coevolving populations of bacteriophage λ and Escherichia coli, we find conflicting evidence for ARD. Virus‐host infection phenotypes fit the ARD model, yet genomic analyses revealed fluctuating selection dynamics. Rather than coevolution unfolding along a single trajectory, cryptic genetic variation emerges and is maintained at low frequency for generations until it eventually supplants dominant lineages. These observations suggest a hybrid 'leapfrog' dynamic, revealing weaknesses in the predictive power of standard coevolutionary models. The findings shed light on the mechanisms that structure coevolving ecological networks and reveal the limits of using phenotypic or genomic data alone to differentiate coevolutionary dynamics. [ABSTRACT FROM AUTHOR]

Antibiotic resistance is one of the major challenges that humankind shall face in the short term. (Bacterio)phage therapy is a valuable therapeutic alternative to antibiotics and, although the concept is almost as old as the discovery of phages, its wide application was hindered in the West by the discovery and development of antibiotics in the mid-twentieth century. However, research on phage therapy is currently experiencing a renaissance due to the antimicrobial resistance problem. Some countries are already adopting new ad hoc regulations to favor the short-term implantation of phage therapy in clinical practice. In this regard, the Phage Therapy Work Group from FAGOMA (Spanish Network of Bacteriophages and Transducing Elements) recently contacted the Spanish Drugs and Medical Devices Agency (AEMPS) to promote the regulation of phage therapy in Spain. As a result, FAGOMA was asked to provide a general view on key issues regarding phage therapy legislation. This review comes as the culmination of the FAGOMA initiative and aims at appropriately informing the regulatory debate on phage therapy. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Typhimurium (S. Typhimurium) elicits the starvation-stress response (SSR) due to starvation for an essential nutrient, e.g. a carbon/energy source (C-source). As part of the SSR, the alternative sigma factor σE is activated and induced. The authors suspect that this activation is, in part, triggered by changes in the S. Typhimurium cell envelope occurring during the adaptation from growth to carbon/energy starvation (C-starvation), and resulting in an increased need for σE-regulated factors involved in the proper folding and assembly of newly synthesized proteins destined for this extracytoplasmic compartment. This led to the hypothesis that a σE activation signal might arise during C-source shifts that cause the induction of proteins localized to the extracytoplasmic compartment, i.e. the outer membrane or periplasm, of the cell. To test this hypothesis, cultures were grown in minimal medium containing enough glucose to reach mid-exponential-phase, plus a non-limiting amount of a secondary ‘less-preferred’ but utilizable carbon/energy source. The σE activity was then monitored using plasmids carrying rpoEP1- and rpoEP2-tacZ transcriptional fusions, which exhibit σE-independent and -dependent lacZ expression, respectively. The secondary C-sources maltose, succinate and citrate, which have extracytoplasmic components involved in their utilization (e.g. LamB), resulted in a discernible diauxic lag period and a sustained increase in σE activity. Growth transition from glucose to other utilizable phosphotransferase (PTS) and non-PTS C-sources, such as trehalose, mannose, mannitol, fructose, glycerol, D-galactose or L-arabinose, did not cause a discernible diauxic lag period or a sustained increase in σE activity. Interestingly, a shift from glucose to melibiose, which does not use an extracytoplasmic-localized protein for uptake, did cause an observable diauxic lag period but did not result in a sustained increase in σE activity. In addition, overexpression of LamB from an arabinose-inducible promoter leads to a significant increase in σE activity in the absence of a glucose to maltose shift or C-starvation. Furthermore, a Δlamb::Ω-Kmr mutant, lacking the LamB maltoporin, exhibited an approximately twofold reduction in the sustained σE activity observed during a glucose to maltose shift, again supporting the hypothesis. Interestingly, the LamB protein lacks the typical Y-X-F terminal tripeptide of the OmpC-like peptides that activate DegS protease activity leading to σE activation. It does, however, possess a terminal pentapeptide (Q-M-E-I-W-W) that may function as a ligand for a putative class II PDZ-binding site. The authors therefore propose that the σE regulon of S. Typhimurium not only is induced in response to deleterious environmental conditions, but also plays a role in the adaptation of cells to new growth conditions that necessitate changes in the extracytoplasmic compartment of the cell, which may involve alternative signal recognition and activation pathways that are independent of DegS. [ABSTRACT FROM AUTHOR]

For the protection of Protected Geographical Indication (PGI) Sunite lamb, PGI Sunite lamb samples and lamb samples from two other banners in the Inner Mongolia autonomous region were distinguished by stable isotopes (δ13C, δ15N, δ2H, and δ18O) and two local modeling approaches. In terms of the main characteristics and predictive performance, local modeling was better than global modeling. The accuracies of five local models (LDA, RF, SVM, BPNN, and KNN) obtained by the Adaptive Kennard–Stone algorithm were 91.30%, 95.65%, 91.30%, 100%, and 91.30%, respectively. The accuracies of the five local models obtained by an approach of PCA–Full distance based on DD–SIMCA were 91.30%, 91.30%, 91.30%, 100%, and 95.65%, respectively. The accuracies of the five global models were 91.30%, 91.30%, 91.30%, 100%, and 91.30%, respectively. Stable isotope ratio analysis combined with local modeling can be used as an effective indicator for protecting PGI Sunite lamb. [ABSTRACT FROM AUTHOR]

In this study, Klebsiella pneumoniae was suspended in synthetic saliva in a nebulizer (N0) and nebulized for 5 min (N5) into an aerosol chamber and further prolonged in the aerosolization phase for 15 min (A15) under four different conditions: 20°C, 50% relative humidity (RH); 20°C, 80% RH; 30°C, 50% RH; and 30°C, 80% RH. Samples were collected at N0, N5, and A15, then subjected to survival analysis and comparative transcriptomic analysis in order to help elucidate the underlying mechanisms of airborne survival. Survival analysis shows that a higher humidity and lower temperature were favorable for the airborne survival of K. pneumoniae, and the effect of RH was more remarkable at 20°C than that at 30°C. The RNA‐seq results show that during the nebulization phase (N0 vs. N5), a total number of 201 differentially expressed genes (DEGs) were identified (103 downregulated and 98 upregulated). Comparison between nebulization and aerosolization phases (N5 vs. A15) indicates up to 132 DEGs, with 46 downregulated and 86 upregulated. The most notable groups of genes are those involved in cellular remodeling, metabolism and energy processes. Alarmingly, the mbl gene, which encodes antibiotic resistance in K. pneumoniae, was upregulated during the suspension phase under all the tested conditions. This study provides insights into the control of airborne transmitted diseases. [ABSTRACT FROM AUTHOR]

There is a growing consensus that traits offer a powerful way to examine the relationship between the environment, organismal strategies, species interactions, and ecological success. To date, trait‐based research has largely been focusing on individual trophic levels and not on cross‐level interactions. Looking at traits not only within but across trophic levels and identifying traits that together define trophic interactions holds a great potential for understanding the mechanisms of interactions. Here, we outline the conceptual foundation for cross‐trophic trait‐based frameworks, using planktonic food webs as an example. First, we compile a list of traits important within different individual trophic levels and show that there are traits that are common across trophic levels ("universal" traits), as well as trophic level‐specific traits. Next, we focus on traits that characterize interactions across trophic levels, focusing on two types of interaction—grazer–primary producer and host–parasite, identifying the similarities and differences between these interactions. We outline the trait hierarchies that define possible and realized intertrophic interactions and their strengths. We then highlight the importance of trade‐offs among those traits in shaping interactions and explaining general patterns in the structure and function of food webs. Finally, we discuss the environmental influences on traits, their eco‐evolutionary responses to changing conditions and how those responses may alter trophic interactions. The extension of trait‐based approaches from individual trophic levels to food webs and different trophic interactions should stimulate further conceptual development, enrich the field of aquatic sciences, and provide a framework to better predict global change effects on ecosystems. [ABSTRACT FROM AUTHOR]

Forage allowance impacts dry matter (DM) intake and the use of nutrients by ruminants. The efficient use of protein and energy from pasture is related to better livestock performance and lower environmental impacts. The aims of this study were to evaluate the effect of forage allowance levels on intake, digestibility, nitrogen (N) and energy balance, and methane (CH4) emissions by lambs fed fresh pearl millet [Pennisetum americanum (L.) Leeke]. An indoor trial was performed using lambs in a completely randomized design with four treatments [forage allowance at 1.5, 2.0, 2.5 kg DM/100 kg of live weight (LW), and ad libitum allowing 20% of refusals] and four replicates (lambs). Forage intake, digestibility, total urine and feces excretion, and CH4 emission were measured to calculate N and energy balances. An increase in forage allowance resulted in a linear increase in lamb forage intake, N retention, and metabolizable energy intake. Moreover, lamb CH4 emission (g/day) also increased with greater forage allowance, while CH4 yield decreased linearly as forage allowance increased. Our results indicate that maximizing forage intake improves N and energy use efficiency and mitigates CH4 yield and decreases CH4 conversion factor (Ym) by lambs fed pearl millet forage. Thus, management strategies that optimize intake of tropical forages by ruminants improve the use of nutrients ingested and mitigates negative impacts to the environment. [ABSTRACT FROM AUTHOR]

The application of probiotics, in aquaculture, is becoming increasingly widespread and have had positive application effects. However, reports of loach-derived probiotics are quite limited. In this study, two representative strains of lactic acid bacteria with excellent traits, namely, Weissella confusa N17 and Lactobacillus saniviri N19, were screened from the intestine of healthy loaches. W. confusa N17 and L. saniviri N19 could inhibit different common various pathogenic bacteria, especially Aeromonas spp., and were sensitive to the most common antibiotics. The survival rate of the two strains exceeded 50% after 4 h of incubation in 10% loach bile. Moreover, the two strains showed significant tolerance to trypsin. Their autoaggregation capacity and hydrophobicity were greater than 30%. In addition, the aggregation ability of both strains was higher than 30% for both A. veronii TH0426 and A. hydrophila TPS. The two strains had a high biofilm-forming ability and strong adhesion to epithelioma papulosum cyprini (EPC) cells. Scanning electron microscopy results showed that the culture supernatants of the two strains had a significantly destructive effect on A. veronii TH0426 and A. hydrophila TPS. Overall, the traits of W. confusa N17 were better than those of L. saniviri N19. Genome sequencing and analysis demonstrated a lack of virulence factor-related or drug resistance-related genes in genome N17. The diet supplemented with the W. confusa N17 strain significantly improved the resistance of loaches to A. veronii infection, and the protection rate reached 57.1%. Therefore, W. confusa N17 exhibits promising for further applications in loach aquaculture., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Recent studies applying advanced imaging techniques are changing the way we understand bacterial cell surfaces, bringing new knowledge on everything from single-cell heterogeneity in bacterial populations to their drug sensitivity and mechanisms of antimicrobial resistance. In both Gram-positive and Gram-negative bacteria, the outermost surface of the bacterial cell is being imaged at nanoscale; as a result, topographical maps of bacterial cell surfaces can be constructed, revealing distinct zones and specific features that might uniquely identify each cell in a population. Functionally defined assembly precincts for protein insertion into the membrane have been mapped at nanoscale, and equivalent lipid-assembly precincts are suggested from discrete lipopolysaccharide patches. As we review here, particularly for Gram-negative bacteria, the applications of various modalities of nanoscale imaging are reawakening our curiosity about what is conceptually a 3D cell surface landscape: what it looks like, how it is made and how it provides resilience to respond to environmental impacts., (© 2023. Springer Nature Limited.)

Deals with a study that examined the metal binding properties of Escherichia coli (E. coli) strains displaying short peptides as a fusion to maltoporin (LamB) protein. Stoichiometry of cadmium binding to synthetic peptides; Expression of LamB hybrid proteins; Metal binding properties of E. coli displaying LamB-MBS.

Presents a study regarding the interaction of the signal sequence of the Escherichia coli LamB protein with various lipid bilayers with the use of circular dichroism, tryptophan fluorescence, electrophoretic mobility, dipole potential, and binding measurements. Synthesis of three LamB signal peptide differing in their net charge; What microelectrophoresis experiments revealed; Reference to the consolving of lipids.

LamB (maltoporin) of Salmonella typhimurium was found to be more strongly associated with the murein than OmpF. It was purified in one step using a hydroxyapatite (HTP) column. Reconstitution of the pure protein with lipid bilayer membrane showed that LamB of S. typhimurium formed small ion-permeable channels with a single channel conductance of about 90pS in 1M KCI and some preference for cations over anions. The conductance concentration curve was linear, which suggested that LamB of S. typhimurium does not contain any binding site for ions. Pore conductance was completely inhibited by the addition of 20 mM maltotriose. Titration of the LamB-induced membrane conductance with different sugars, including all members of the maltooligosaccharide series up to seven glucose residues, suggested that the channel contains, like LamB (maltoporin) of Escherichia coli, a binding site for sugars. The binding constant of sugars of the maltooligosaccharide series increased with increasing number of glucose residues up to five (saturated). Small sugars had a higher stability constant for sugar binding relative to LamB of E. coli. The advantage of a binding site inside a specific porin for the permeation of solutes is discussed with respect to the properties of a general diffusion porin. [ABSTRACT FROM AUTHOR]

After fusion with the N-proximal portion of the outer membrane protein LamB, three β-adrenergic receptors, the human β1- and β2- and turkey β1-adrenergic receptor, were expressed in Escherichia coli with retention of their own specific pharmacological properties. Molecular characterization and localization of the three receptors in bacteria and comparison of the behaviour of each hybrid protein are reported. The bacteria were lysed and fractionated on a sucrose gradient. Saturable [125I]iodocyanopindolol binding activity was found associated mainly with the inner membrane fraction, suggesting that the receptor is correctly folded in this membrane. Binding activity was also found in the outer membrane fraction but varied according to the receptor type. Photoaffinity labeling experiments revealed that the receptors exhibit binding activity only after proteolytic removal of the LamB moiety from the fusion protein. The three hybrid proteins, detected in immunoblots by anti-peptide antibodies, were found mainly in the outer membrane fraction. Each of them exhibited different susceptibility to intrinsic bacterial proteolytic enzymes; sites of proteolytic cleavage were localized by the use of anti-peptide antibodies. The functional expression in E. coli of three β-adrenergic receptors with similar structure but different amino acid sequences suggests that this expression system may be a general feature among similar receptors of the family of G-protein-coupled receptors. The level of expressed binding activity of a given receptor will be within the control of proteolytic degradation processes, depending on the primary sequence of the receptor. Constructions of new hybrid proteins, in combination with expression in protease mutants of E. coli, should help in controlling such processes. [ABSTRACT FROM AUTHOR]

Presents the genetics of active transport in bacteria. Inclusion of the fundamental processes of all cells; Location of functionally important sites for substrate recognition and energy coupling; Conformational changes that occur during the transfer of substrate from one side of the membrane to the other.

Summary: In the study, lamb, feed and soil samples were collected from three regions of China in the year 2017 and 2018. The δ13C and δ15N values in lamb (defatted dry mass and lipids) and grass and the contents of nine elements in lamb, grass and soil were determined. The results indicated that except for Fe, Zn and Rb, most variables of lamb showed significant differences among two or three regions, and the contents of Fe, Sr and Cs showed significant differences between two years, and the contents of Cu and Zn showed significant differences among different muscle sections. Besides, significant correlations were found for δ13C between defatted lamb and lipid (r2 = 0.973, P < 0.05) and for the element contents between lamb and soil. Finally, four key variables δ13C, δ15N, Sr and Mo were selected through stepwise canonical discriminant analysis with the cross‐validation correct classification rate of 99.1%. [ABSTRACT FROM AUTHOR]

Aeromonas veronii (A. veronii) is one of the important zoonotic pathogens. The development of drug resistance and enrichment of resistant genes not only cause misery from bacterial diseases but also pose a threat to public health security. In this study, A. veronii was isolated and identified from 357 shellfish samples imported from 6 aquaculture bases in Jilin Province, China, and tested for virulence, tolerance and drug resistance genes. According to 16S rRNA and gyrB gene PCR result, 47 strains were identified as Aeromonas veronii. The distribution of 9 virulence factors including fla (55.3%), Alt (42.5%), Act (31.9%), Aer (31.9%), Lip (19.1%), gcaT (12.7%), ahyB (12.7%), ascV (10.6%) and Ast (8.5%) was determined exclusively in Aeromonas veronii. Nine virulence genes were detected in two isolates. In addition, the resistance rate of isolates to sulfamonomethoxine and florfenicol was 100%, while to tetracycline, aureomycin and doxycycline it was 34.0%(16/47),29.8%(14/47)and 25.5%(12/47), respectively. Among all tested strains, thirty-five isolates were sensitive to enrofloxacin, and the majority of drug-resistant strains carried strA(95.7%), strB (93.6%), qnrA(40.4%), qnrD(31.9%), tetQ (31.9%)and tetK (29.8%) resistance genes. The results provide a basis for a risk assessment of the drug resistance of Aeromonas veronii. [ABSTRACT FROM AUTHOR]

Aeromonas veronii is an important zoonotic and aquatic pathogen. An increasing number of reports indicate that it has caused substantial economic losses in the aquaculture industry, in addition to threatening human health. However, little is known about its pathogenesis. Exploration of new virulence factors of A. veronii would be helpful for further understanding its pathogenesis. Hence, we comparatively analyzed the proteomes of virulent, attenuated, and avirulent strains of A. veronii using tandem mass tag (TMT) protein labeling and found numerous proteins either up‐ or downregulated in the virulent strain. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses showed that these differentially expressed proteins (DEPs) were involved mainly in pathways associated with bacterial chemotaxis and microbial metabolism in diverse environments. Furthermore, the expression levels of lysine decarboxylase, endoribonuclease, maltoporin, pullulanase, and aerolysin were positively correlated with the virulence of the strains, suggesting that their function may be closely related to the virulence of A. veronii. The results of qRT‐PCR and multiple reaction monitoring for some DEPs were consistent with the results of TMT protein labeling. These results suggest that these DEPs may be novel potential virulence factors and will help to further understand the pathogenesis of A. veronii. [ABSTRACT FROM AUTHOR]

Background: Microbial evolution experiments can be used to study the tempo and dynamics of evolutionary change in asexual populations, founded from single clones and growing into large populations with multiple clonal lineages. High-throughput sequencing can be used to catalog de novo mutations as potential targets of selection, determine in which lineages they arise, and track the fates of those lineages. Here, we describe a long-term experimental evolution study to identify targets of selection and to determine when, where, and how often those targets are hit. Results: We experimentally evolved replicate Escherichia coli populations that originated from a mutator/nonsense suppressor ancestor under glucose limitation for between 300 and 500 generations. Whole-genome, whole-population sequencing enabled us to catalog 3346 de novo mutations that reached > 1% frequency. We sequenced the genomes of 96 clones from each population when allelic diversity was greatest in order to establish whether mutations were in the same or different lineages and to depict lineage dynamics. Operon-specific mutations that enhance glucose uptake were the first to rise to high frequency, followed by global regulatory mutations. Mutations related to energy conservation, membrane biogenesis, and mitigating the impact of nonsense mutations, both ancestral and derived, arose later. New alleles were confined to relatively few loci, with many instances of identical mutations arising independently in multiple lineages, among and within replicate populations. However, most never exceeded 10% in frequency and were at a lower frequency at the end of the experiment than at their maxima, indicating clonal interference. Many alleles mapped to key structures within the proteins that they mutated, providing insight into their functional consequences. Conclusions: Overall, we find that when mutational input is increased by an ancestral defect in DNA repair, the spectrum of high-frequency beneficial mutations in a simple, constant resource-limited environment is narrow, resulting in extreme parallelism where many adaptive mutations arise but few ever go to fixation. [ABSTRACT FROM AUTHOR]

Vibriosis is a commonly found bacterial disease identified among fish and shellfish cultured in saline waters. A multitude of Vibrio species have been identified as the causative agents. LamB, a member of outer membrane protein (OMPs) family of these bacteria is conserved among all Vibrio species and has been identified as an efficient vaccine candidate against vibriosis. Rootless duckweed (Wolffia) is a tiny, edible aquatic plant possessing characteristics suitable for the utilization as a bioreactor. Thus, we attempted to express a protective edible vaccine antigen against fish vibriosis in nuclear-transformed Wolffia. We amplified LamB gene from virulent Vibrio alginolyticus and it was modified to maximize the protein expression level and translocate the protein to the endoplasmic reticulum (ER) in plants. It was cloned into binary vector pMYC under the control of CaMV 35S promoter and introduced into Wolffia globosa by Agrobacterium-mediated transformation. Integration and expression of the LamB gene was confirmed by genomic PCR and RT-PCR. Western blot analysis revealed accumulation of the LamB protein in 8 transgenic lines. The cross-protective property of transgenic Wolffia was evaluated by orally vaccinating zebrafish through feeding fresh transgenic Wolffia and subsequently challenging with virulent V. alginolyticus. High relative percent survival (RPS) of the vaccinated fish (63.3%) confirmed that fish immunized with transgenic Wolffia were well-protected from Vibrio infection. These findings suggest that Wolffia expressed LamB could serve as an edible plant-based candidate vaccine model for fish vibriosis and feasibility of utilizing Wolffia as bioreactor to produce edible vaccines.

Many gene therapy strategies require transfer of highmolecular weight DNA into human cells. To enable clinical trials, these vectors need to be produced on a large scale and at low cost. The production of effective high-capacity vectors like HSV-amplicons and helperdependent adenoviral vectors is difficult to up-scale, so new inexpensive vectors are needed for the efficient delivery of high-molecular weight DNA to human cells. Bacteriophage lambda vectors can accommodate up to about 46 kb of therapeutic DNA and can be easily produced in an industrial setting. However, the lambda vectors transfer DNA into mammalian cells with only a low efficiency. It was shown that bacteriophage lambda virions ejected their DNA in the presence of the purified receptor for bacteriophage lambda, maltoporin (LamB protein), encoded by the malB gene of Shigella sonnei 3070. This property of S. sonnei maltoporin was exploited for the bacteriophage injection-driven DNA loading of liposomes and other polymer nanocontainers displaying maltoporin. Relying on the above evidence I hypothesize that the efficient gene transfer by industrially produced bacteriophage lambda vector virions, such as cosmid transducing particles, to human cells can be accomplished after incorporation (protein painting) of the purified S. sonnei maltoporin into the human plasma membrane. [ABSTRACT FROM AUTHOR]

The purpose of this study was to compare the in vitro digestibility levels and chemical compositions of commercial extruded dry-type adult dog foods with different types of protein contents [fish meat (F-dog foods) (n = 7), lamb meat (L-dog foods) (n = 9), or poultry meat (P-dog foods) (n = 8)]. The in vitro digestion values of premium commercial dog foods were examined at three stages: gastric digestion, small intestine digestion and large intestine digestion/fermentation. The metabolisable energy (ME), crude protein (CP), starch, diethyl ether extract (EE) and ash contents and the in vitro cumulative gas production values of all the premium dog foods differed significantly among the commercial brands in the same category (F-, L- or P-dog foods) (P < 0.05). The crude fibre (CF) and the CP/1 000 kcal ME values of the F- and P-dog foods demonstrated a significant difference among the commercial brands (P < 0.05). The organic matter disappearance (OMd) values of the L-dog foods showed a significant difference among the commercial brands (P < 0.05); but the OMd values of the F- and P-dog foods did not differ among the commercial brands (P > 0.05). The average values of the OMd for the F-dog foods were more rapid than the average for the L- and P-dog foods, in the evaluation of all the foods (P = 0.001). Besides, the price of the L-dog foods was positively correlated with the OMd and CP of the L-dog foods; however, it was negatively correlated with the NFE (nitrogen free extract) and CHO (total carbohydrates) of the L-dog foods (P < 0.05). The CP values of the L-dog foods were positively correlated with the OMd values (P < 0.05). Although price is an important determinant of food quality in the L-dog foods, it is not in the F- and P-dog foods. In the general evaluation of all the dog foods, there was no correlation among the food price and the digestibility and the nutrient content for all of the premium dog foods. The present study indicated that the energy, nutrient matter and digestibility of premium dog foods changed with the change in the variety and the amount of the feedstuffs. The digestibility of the dog foods with the fish meat were higher than those of the other dog foods. The amount of protein that an adult dog will receive with 1 000 kcal of DM (dry matter) consumption of premium dog foods with fish meat and chicken meat, varied among the brands. This point showed the need to pay attention to the food consumption amount of the dogs and the energy-protein balance in their diets, especially dog foods with fish meat and chicken meat. [ABSTRACT FROM AUTHOR]