Antimicrobial-resistant bacteria pose serious public health risks, necessitating bioprospecting for novel antimicrobial drugs. The endophytic fungi of the mangrove ecosystem are hotspots for discovering new bioactive chemical compounds. In this context, an investigation was designed to determine the isolation of the major endophytic fungi inhabiting the leaves of Acanthus ilicifolius, a mangrove plant with a long history of traditional use in the Chinese and Indian medical systems. Based on the morphological characterizations and molecular analysis of internal transcribed spacer rDNA sequence data, the study identified three unique endophytic fungal species, namely, Periconia macrospinosa, Coprinopsis cinerea, and Alternaria sp. Of these, P. macrospinosa was identified as the most dominant one, with the highest relative frequency (35.22%). The antibacterial activity of P. macrospinosa isolate (CMFRI/fPM-01) was evaluated by the well and diffusion method against six human pathogens, viz., Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus. The results demonstrated a high and wide spectrum of antimicrobial action of the isolate against all the tested human pathogens, with no significant difference (P > 0.05) in the activity between the pathogens. The antibacterial activity was further confirmed by determining the fungal culture supernatant's minimum inhibitory concentration and minimum bactericidal concentration. Although the studied fungi are known from other sources, this is the first report of P. macrospinosa and C. cinerea as endophytes in A. ilicifolius leaves. The outcomes also showed that the P. macrospinosa isolate could be used to discover effective antibacterial drugs against various human diseases.

Due to the environmental problems derived from the use of common surfactants as modifiers for clay mineral adsorbents to mitigate mycotoxin contamination of animal feeds, finding non-toxic modifiers to prepare safe and efficient adsorbents is necessary. The objective of the present study was, therefore, to modify acidified palygorskite with polyhexamethylene biguanide (PHMB) to obtain antibacterial polyhexamethylene biguanide/palygorskite (PHMB/Plg) composites for the removal of zearalenone, a common mycotoxin. The PHMB/Plg composites were characterized and analyzed by X-ray diffraction, Fourier-transform infrared spectroscopy, field-emission scanning electron microscopy, and isothermal nitrogen adsorption analysis. The adsorption properties of the composites with respect to zearalenone and their antibacterial activity with respect to Escherichia coli and Staphylococcus aureus were studied. The results indicated that the hydrophobicity of palygorskite was enhanced after modification with PHMB, which could effectively improve the adsorption property of palygorskite toward the nonpolar zearalenone molecules. The adsorption capacity of PHMB/Plg increased with increasing amounts of polyhexamethylene biguanide and increasing pH. The adsorption data were described well by pseudo-second order kinetics and by the Langmuir adsorption model. The maximum adsorption capacity was 2777 μg/g. When the amount of PHMB added increased to 15 wt.%, the composites obtained exhibited good antibacterial performance, and the minimum inhibitory concentrations for Escherichia coli and Staphylococcus aureus were both at 2.5 mg/mL.

The immobilization of antimicrobial drugs can be used to expand the application of antibacterial properties to consumer products. The purpose of this study was to stabilize an antimicrobial agent, levofloxacin (LVX), for sustained antibacterial activity by immobilizing the drug molecules in a layered double hydroxide (LDH) and embedded in a polyurethane substrate. As-prepared MgAl-LDH was calcined at 400°C and reconstructed with LVX for intercalation. The X-ray diffraction patterns and cross-sectional transmission electron microscopy images showed lattice expansion along the crystallographic c axis upon LVX intercalation, suggesting successful loading of the drug. Fourier-transform infrared spectra revealed that the structure of LVX was well preserved between LDH layers. Elemental analysis indicated that the loading capacity of LVX in the hybrid was 41.7%. Bacterial-colony forming inhibitory assay on Bacillus subtilis exhibited ~100% antibacterial activity of both LVX alone and LVX-LDH hybrid (LL). To determine sustainability of antibacterial activity by the hybrid, either LVX alone or LL hybrid was loaded in the polyurethane (PU) substrate for which antibacterial activity was evaluated before and after immersion in a phosphate-buffered saline for 3 days. The LVX-composited PU showed a dramatic decrease in antibacterial activity, down to 0% after buffer treatment; LL-composited PU still contained antibacterial activity (~34% of colony suppression) after phosphate-buffered saline immersion.

Supported silver nanoparticles (Ag NPs) have been used extensively as antibacterial agents in biomedicine, biotechnology, and environmental remediation. However, a facile and scalable method for preparing Ag NPs dispersed homogeneously on supports remains a challenge. In this study, a novel molten salt method was developed successfully to synthesize the supported, homogeneously dispersed Ag NPs on palygorskite. Abundant pores and ample surface hydroxyl groups of palygorskite served as anchoring sites, preventing the rapid growth, aggregation, and sintering of Ag NPs. Typically, palygorskite was mixed with AgNO3 (as a precursor) and NaNO3 (as a dispersant), and then the mixture was heated slowly. During the heating process, the AgNO3 decomposed gradually into Ag NPs and the molten NaNO3 with a high concentration of ions dispersed the newly formed Ag NPs. The Ag NPs were dispersed homogeneously on the palygorskite and had very small particle sizes (~5.8 nm) even for a significant loading amount (~9 wt.%). As antibacterial agents, the Ag/palygorskite nanocomposites showed enhanced antibacterial activity, compared with those synthesized without the introduction of molten NaNO3. In addition, the key effect of the surface hydroxyl groups of palygorskite on the characteristics of the loaded Ag and the corresponding antibacterial activity were also elucidated. As such, the present work provided a novel and facile strategy for the synthesis, without a chemical reductant or surfactant, of supported, highly dispersed Ag NPs on clay minerals and this could have potential in the scalable production and practical application of Ag-based antibacterial materials.

The present study introduces an overview of gentamicin-clay mineral systems for applications in biomedicine and then focuses on the development of a series of gentamicin/clay hybrid materials to be used as the bioactive phase of hydroxypropylmethylcellulose (HPMC) to produce bionanocomposite membranes possessing antimicrobial activity of interest in wound-dressing applications. Gentamicin (Gt) was adsorbed from aqueous solutions into a montmorillonite (Cloisite®-Na+) to produce intercalation compounds with tunable content of the antibiotic. The hybrids were characterized by CHN chemical analysis, energy-dispersive X-ray analysis, X-ray diffraction, Fourier-transform infrared spectroscopy, and thermogravimetric analysis, confirming the intercalation of Gt by an ion-exchange mechanism. The release of Gt from the hybrids was tested in water and in buffer solution to check their stability. Hybrids with various amounts of Gt were incorporated into a HPMC matrix at various loadings and processed as films by the casting method. The resulting Gt-clay/HPMC bionanocomposites were characterized by means of field-emission scanning electron microscopy, and were also evaluated for their water-adsorption and mechanical properties to confirm their suitability for wound-dressing applications. The antimicrobial activity of the bionanocomposite films was tested in vitro toward various microorganisms (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium, Acinetobacter baumannii, and Klebsiella pneumonia), showing a complete bacterial reduction even in films with small Gt contents.

This research paper aimed to examine the antibacterial activity of lactoferrin (LF) as a potential natural alternative in the dairy sector, by measuring its minimum inhibitory concentration (MIC) against a number of common food-borne pathogens as well as Pseudomonas aeruginosa, one of the major dairy product spoiling microorganisms. Additionally, a viability experiment was applied to laboratory-manufactured set yoghurt to assess its impact on the activity of starter culture, sensory properties and STEC survivability. The findings demonstrated that LF exhibited significant antimicrobial activity, particularly against E. coli and S. typhimurium with MIC values of 0.0001 and 0.01 mg/ml, respectively. However, P. aeruginosa and B. cereus were quite resistant to LF requiring higher concentrations for MIC (2.5 mg/ml). By the third day of storage, LF at 0.0001 and 0.001 mg/ml significantly reduced the survivability of Shiga toxin-producing E. coli STEC by 70 and 91.6%, respectively, in the lab-manufactured yoghurt. Furthermore, LF enhanced the sensory properties of fortified yoghurt with a statistically significant difference in comparison to the control yoghurt group. There was no interference with the activity of the starter culture throughout the manufacturing process and the storage period. In conclusion, the potent antimicrobial effect of LF opens a new avenue for the dairy industry's potential applications of LF as a natural preservative without negatively influencing the sensory properties and starter culture activity of fermented products.

Novel microencapsulated n-octadecane with natural silk fibroin (SF) shell attached with silver nanoparticles (AgNPs) on its surface was synthesized in oil-in-water emulsion via a self-assembly method. No additional reductant was used in the in situ preparation of AgNPs due to the inherent reduction property of tyrosine (Tyr) residues in SF. The microstructures and particle sizes of the resultant microcapsules were investigated by using a scanning electron microscope (SEM) and a laser scattering particle size distribution analyzer. The resulting microcapsules exhibited a regular spherical morphology with a 4–5 μm narrow diameter distribution range. And the AgNPs attached to the surface exhibited an even distribution. According to the analytical results of DSC, TGA, and infrared system, the SF-AgNPs microcapsule presents enhanced thermal stability and obvious thermal regulation properties. In addition, it was found that the SF-AgNP microcapsule also exhibited a good antibacterial activity against both Gram-positive bacteria (Staphylococcus aureus), and Gram-negative bacteria (Escherichia coli). The SF-AgNPs microcapsule synthesized in this study could be a potential candidate for thermal regulation and healthcare applications.

A superelastic Ti–40Nb alloy enhanced with Cu element (0, 2.5, 5, 7.5, and 10 wt%) was synthesized by a spark plasma sintering method to obtain biomaterials with an antimicrobial effect. The microstructure results showed that β phase was the main phase in (Ti–40Nb)–Cu alloys while Ti2Cu was synthesized with the Cu addition above 5 wt%. (Ti–40Nb)–Cu alloys exhibited high compressive strength over 1693.08 MPa, high yield strength of 1140.26–1619.14 MPa, low elastic modulus in the range of 43.91–58.01 GPa, low elastic energy (14.81–24.73 MJ/m3), and together with large plastic strain over 18.5%. High concentration of Cu ion released steadily from alloys in early 7 days, then the released concentration of Cu ion showed long-lasting and moderate. Comparing with the Ti–40Nb alloy, high antimicrobial activity was pronounced on (Ti–40Nb)–Cu alloys, and (Ti–40Nb)–Cu alloys showed more inhibitory activity against bacteria (E. coli and S. aureus) than fungi (C. albicans). Cu contents in alloys influenced the Cu ion release, which in turn affected the antimicrobial activity. As a good combination of low elastic modulus, high mechanical properties, good elastic energy, and excellent antimicrobial performance, (Ti–40Nb)–Cu alloys offer potential advantages to prevent stress shielding and exhibit an excellent antimicrobial property for hard tissue replacements.

The development of synthetic scaffolds with a desirable combination of properties, such as bioactivity, the ability to locally deliver antibacterial agents and high osteogenic capacity, is a challenging but promising approach in bone tissue engineering. In this study, scaffolds of a borosilicate bioactive glass (composition: 6Na2O, 8K2O, 8MgO, 22CaO, 36B2O3, 18SiO2, 2P2O5; mol%) with controllable antibacterial activity were developed by doping the parent glass with varying amounts of Ag2O (0.05, 0.5, and 1.0 wt%). The addition of the Ag2O lowered the compressive strength and degradation of the bioactive glass scaffolds but it did not affect the formation of hydroxyapatite on the surface of the glass as determined by energy dispersive x-ray analysis, x-ray diffraction, and Fourier transform infrared analysis. The Ag2O-doped scaffolds showed a sustained release of Ag ions over more than 8 weeks in simulated body fluid and resistance against colonization by the bacterial strains Escherichia coli and Staphylococcus aureus. In vitro cell culture showed better adhesion, proliferation, and alkaline phosphatase activity of murine osteoblastic MC3T3-E1 cells on the Ag2O-doped bioactive glass scaffolds than on the undoped scaffolds. The results indicate that these Ag-doped borosilicate bioactive glass scaffolds may have potential in repairing bone coupled with providing a lower risk of bacterial infection.

Because immune defenses are often costly employed, insect immunocompetence cannot be always maintained at its maximum level. Here, the oriental fruit fly, Bactrocera dorsalis (Hendel), was used as a study object to investigate how its immune defenses varied with the developmental stage change and mating access. Our data indicated that both phenoloxidase (PO) activity and antibacterial activity significantly increased from new larvae to pupae but decreased in adults after emergence. Furthermore, both the PO activity and antibacterial activity in the hemolymph of copulated male and female adults were dramatically higher than that of virgin male and female ones, respectively. It provided the evidence that copulation could increase the magnitude of immune defense in hemolymph of B. dorsalis. Together, these results suggest that B. dorsalis possess a flexible investment strategy in immunity to meet its specific needs based on the endo- and exogenous factors, such as their distinct food source and living environments.

Rhynchophorus ferrugineus (Coleoptera, Curculionidae) is the most destructive pest of palm trees worldwide containing it invasive areas, such as the southern part of China. It is always emphasized to develop integrated pest management based on biological agents, but their success is not very exciting. Presently, the immune defenses of this pest against biological agents attract scarce attention. It is still unclear whether immune priming also generally occurs in insect pests and in response to different pathogens. Our results indicated that previous challenge of bacteria pathogen enhanced the magnitude of phenoloxidase activity and antibacterial activity in R. ferrugineus larvae against the secondary infection. Furthermore, trans-generational immune priming was also determined in this pest, and only challenged R. ferrugineus mothers transferred the immune protection to their offspring which suggested males and females of this pest might have evolved different strategies on the investment of delivering immune protection to their offspring. Importantly, our data provide the evidence to suggest that different kinds of biological control agents might be used alternatively or in combination to fight against R. ferrugineus because of the existence of immune priming with low species-specific level. On the other hand, for this invasive pest, the immune priming may also facilitate its adaptation and dispersal in the new regions.

Vibrio harveyi is the main pathogen of the European abaloneHaliotis tuberculata, and recently caused important mortalities at theproduction sites of this marine gastropod in France. In the present work, the monthlyantibacterial activity of two red seaweed species from the French Atlantic coast, thenative Palmaria palmata and the introduced Grateloupiaturuturu, were investigated against the abalone pathogen Vibrio harveyistrain ORM4. Water-soluble extracts were screened using the microplate method.Grateloupia turuturu showed an antibacterial activity with a maximalgrowth inhibition in spring of around 16%. In contrast, Palmaria palmatawas inactive, as further growth of the bacteria was observed. Preliminaryone-dimensional proton nuclear magnetic-resonance (1H NMR) profiles identified thedifferences between the two water-soluble extracts.

Copper oxide (CuO) nanosheets synthesized in polyvinylpyrrolidone (PVP) were characterized with respect to antimicrobial activity by quick precipitation method. Different sizes and shapes of CuO nanosheets were obtained by simple variations of PVP concentrations. The x-ray diffraction results revealed the formation of pure-phase CuO with monoclinic structure. Transmission electron microscopy analysis showed that the average ratio of length to width of these nanosheets increased with increasing PVP concentrations. Due to the quantum size effect, CuO nanosheets exhibit a blue shift in the ultraviolet-visible spectra. Field emission scanning electron microscopy results showed that as the concentration of PVP increased, well-defined morphologies were formed on the surface of the products. Energy dispersive analysis of x-ray clearly confirmed the presence of Cu and O with an atomic ratio of 1:1. Fourier transform infrared spectroscopy results showed that C=O in PVP coordinated with CuO and formed a protective layer. The mechanism of the reaction was also discussed. CuO nanosheets in suspension showed activity against a range of bacterial pathogens and fungi with minimum bactericidal concentrations (MBCs) ranging from 100 to 5000 µg/mL. The extent of the inhibition zones and the MBCs was found to be size-dependent.

Natural resistance-associated macrophage protein 1 (Nramp1) plays an important role in restraining the growth of intracellular pathogens within macrophages. In this study, Nramp1 cDNA was cloned from Qinchuan cattle and its anti-bacterial activity was demonstrated as being able to significantly inhibit the growth of Salmonella abortusovis and Brucella abortus in macrophages. Calf fibroblasts stably transfected with pSP–NRAMP1–HA vector were used to reconstruct bovine embryos by somatic cell nuclear transfer (SCNT). Reconstructed embryos were maturated in vitro and the blastocyst formation rate (14.0%) was similar to that of control embryos (14.5%). Transgenic blastocysts were transplanted into 43 recipient cattle, of which 14 recipients became pregnant as evidenced by non-return estrus and by rectal palpation. One fetus was aborted after 6½ months of pregnancy and transgene integration was confirmed by semi-quantitative polymerase chain reaction. Together, this study showed that bovine Nramp1 retains biological function against the growth of intracellular bacteria and can be used to reconstruct embryos and produce Nramp1 transgenic cattle, which may benefit the animal and enhance their ability to prevent attack by intracellular pathogens.

The effect of high-pressure treatment (400, 500 and 650 MPa) on the structure and activity of bovine lactoferrin in different iron-saturation forms has been studied by several techniques. The structural changes produced in lactoferrin by high-pressure were analysed by differential scanning calorimetry and fluorescence spectroscopy, and the immunoreactivity by ELISA. The effect of high-pressure was also studied on some biological properties of lactoferrin, such as iron binding capacity, retention of the bound iron, and antibacterial activity against Escherichia coli O157:H7. Results obtained indicate that treatment at 400 MPa does not substantially modify the conformation of lactoferrin, meanwhile treatments at 500 and 650 MPa greatly affect some of its properties. With respect to the antibacterial activity, the apo and native forms of lactoferrin maintain that activity against Esch. coli only after 400 MPa treatment.

French oyster farming has been subject to severe mortalities during the summer months.Results from the research program “Morest”, which ran from 2000 to 2006 and examined thepossible causes of these mortalities, led to the construction of a model to explain theinteraction between environmental factors, oyster physiology and different opportunisticpathogens underlying oyster summer mortality. Temperature, food, reproduction and stresswere the main factors required for oyster mortality. Genetically-based resistance (“R”oysters) or susceptibility (“S” oysters) to summer mortality was revealed by divergentselection. Building on these results, a literature search was made in 2007 on themolecular origin of genetic resistance to such a complex mortality risk. The objectiveswere to lay a foundation for the preparation and orientation of future research directionsand to improve understanding of the underlying physiological mechanism leading to summermortality. Three years later, the resulting conceptual analysis reported here waspresented as an introductory lecture to Physiomar 2010, a conference where many newresults contributing to this research field were also reported. The literature reviewhighlighted two major review articles: the first dealing with nutrition and reproduction(Schneider 2004), the second with reproduction,temperature, oxidative stress and mortality (Heineinger 2002). The effect of nutrition level on energy orientation to growth orreproduction is controlled by endocrine factors. Among these, neuropeptide Y (NPY),ghrelin and leptin neuropeptides appeared to be potential candidates involved in germ-somaorientation in relation to trophic conditions. Depending on reproductive effort andtemperature, a metabolic stress resulting from the germ-soma conflict can appear,characterized by mitochondrial reactive oxygen species (ROS) production. Such an excess ofROS induces perturbations in mitochondrial activity leading to cell death. Many organisms,such as annual plants or the Pacific salmon, do not survive their first reproduction. Incontrast, others increase stress resistance by selection of antioxidant processes(superoxide dismutase SOD, catalase, etc.) through evolution, and survive firstreproduction. A similar difference was observed in the comparison made between R and Soysters, which differed in ROS production, SOD and catalase levels. Such factorscontrolling reproduction and ROS detoxification processes could therefore provide newmarkers for selection of oysters with better resistance to non-specific pathogens,complementing other classic selection approaches against specific pathogens or forimproved immunity. This antioxidant defence mechanism is found in many organisms includingvertebrates and in some invertebrates, including oysters. Its role needs to be consideredin pathology events involving other aquaculture species and it may also contribute toexplaining the increase in marine pathologies under anthropogenic environmental changes.

Human milk samples from three healthy donors were investigated in order to evaluate the antibacterial activity during lactation against Escherichia coli ATCC 25922 and Listeria monocytogenes. The concentration of the main human-milk antimicrobial proteins (lactoferrin (LF), lysozyme (LZ) and secretory immunoglobulin A (sIgA)) was determined by ELISA. Results showed that human milk exhibited antibacterial activity against List. monocytogenes, although it was weakly active against Esch. coli ATCC 25922. The observed antilisterial activity was positively correlated with LZ concentration. In addition, the effect of gastrointestinal proteases, at different pH conditions, that prevail in the stomach of infants (pH 2·0–6·5), on antilisterial activity and protein degradation was evaluated. Hydrolysis with pepsin at pH 4·0–6·5, followed by treatment with pancreatic enzymes, resulted in a decreased hydrolysis of LZ, LF and sIgA and an enhanced antibacterial activity against List. monocytogenes. It is suggested that partial degradation of certain milk proteins at the gastrointestinal level may produce peptides that could act synergistically with the remnant intact proteins.

Lysozyme from buffalo milk was purified to homogeneity and its N-terminal amino acid sequence, biochemical properties and antibacterial spectrum were determined. The purification procedure, comprising ion-exchange chromatography using CM-cellulose and size-exclusion chromatography using Sephadex G-50, conferred 8622-fold purification and 39·3% recovery of lysozyme. The purified enzyme migrated as a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and native PAGE. Immunological purity of lysozyme preparation was confirmed by immuno-electrophoresis. Molecular weight of buffalo-milk lysozyme as determined by SDS-PAGE was 16 kDa and its amino acid composition was determined by reverse phase high performance liquid chromatography (HPLC). The sequence of 23 amino acid residues at the N-terminal end showed 56·5% homology with bovine milk lysozyme and 30·4% with equine milk lysozyme. The specific activity of buffalo milk lysozyme was ten-times that of bovine milk lysozyme. Buffalo-milk lysozyme was active over a wide range of pH and its activity was strongly influenced by molarity of the medium. Antibacterial activity of buffalo-milk lysozyme was determined against 11 species of bacteria; out of seven Gram-positive bacteria tested, four were inhibited, while Gram-negative bacteria were resistant.

Milk is known to provide not only excellent nutrition but also protection against infection in mammalian neonates. Whey proteins, such as immunoglobulins, lactoferrin, lysozyme and lactoperoxidase, are well known antimicrobial components in milk. Håkansson et al. (1995) demonstrated that some multimeric human α-lactalbumin derivatives induced apoptosis in immature and tumour cells. Yoo et al. (1997) found that lactoferricin, which had been isolated from peptic lactoferrin as an antimicrobial peptide, induced apoptosis in tumour and immature mammalian cells. Apoptotic cell death is considered to be an important physiological function in the development and homeostasis of multicellular organisms and to be a highly controlled means of eliminating dangerous, damaged or unnecessary cells without causing tissue damage. Therefore, these findings suggest that whey proteins may have beneficial health effects by restricting specific cell population in the body.