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Animal models of inflammatory bowel disease: lessons from enteric infections.

Mouse models of intestinal inflammation have played a key role in understanding the mechanisms that govern the inflammatory response in the intestine, and in designing new therapeutic strategies in the treatment of patients with inflammatory bowel disease (IBD). Most of these models use chemical challenges, whereas relatively few robust models of intestinal inflammation caused by microbial infection are known. Two common models of infectious murine colitis and typhlitis are infection with the murine epithelial-adherent pathogen, Citrobacter rodentium, and infection of streptomycin-pretreated mice with Salmonella typhimurium. Studies in these models have helped to define the interactions between bacterial pathogens and host immune defenses, thus broadening the understanding of host-microbial interactions in the intestinal tract. Furthermore, such models help to determine the physiologic consequences of neutralizing specific mediators and signaling pathways implicated in inflammation on antimicrobial host defense.

Animals↗

Improvement of soil characteristics and growth of Dorycnium pentaphyllum by amendment with agrowastes and inoculation with AM fungi and/or the yeast Yarowia lipolytica.

The effectiveness of two microbiologically treated agrowastes [dry olive cake (DOC) and/or sugar beet (SB)] on plant growth, soil enzymatic activities and other soil characteristics was determined in a natural soil from a desertified area. Dorycnium pentaphyllum, a legume plant adapted to stress situations, was the test plant to evaluate the effect of inoculation of native arbuscular mycorrhizal (AM) fungi and/or Yarowia lipolytica (a dry soil adapted yeast) on amended and non-amended soils. Plant growth and nutrition, symbiotic developments and soil enzymatic activities were limited in non-amended soil where microbial inoculations did not improve plant development. The lack of nodules formation and AM colonization can explain the limited plant growth in this natural soil. The effectiveness and performance of inocula applied was only evident in amended soils. AM colonization and spores number in natural soil were increased by amendments and the inoculation with Y. lipolytica promoted this value. The effect of the inoculations on plant N-acquisition was only important in AM-inoculated plants growing in SB medium. Enzymatic activities as urease and protease activities were particularly increased in DOC amended soil meanwhile dehydrogenase activity was greatest in treatments inoculated with Y. lipolytica in SB added soil. The biological activities in rhizosphere of agrowaste amended soil, used as indices of changes in soil properties and fertility, were affected not only by the nature of amendments but also by the inoculant applied. All these results show that the lignocellulosic agrowastes treated with a selected microorganism and its further interaction with beneficial microbial groups (native AM fungi and/or Y. lipolytica) is a useful tool to modify soil physico-chemical, biological and fertility properties that enhance the plant performance probably by making nutrients more available to plants.

Agriculture↗

The causes of and cures for contact lens-induced peripheral ulcer.

PURPOSE: Contact lens-induced peripheral ulceration (CLPU) is a relatively common adverse response associated with wearing hydrogel lenses, especially on an extended wear schedule. Bacteriologic examination of lenses at the time of an event has demonstrated an association with Staphylococci spp. We sought to investigate the causes of CLPU in a rabbit model of contact lens wear. METHODS: Rabbits wore contact lenses for a period of 24 hr in the presence or absence of bacteria or in presence or absence of epithelial scratches made in the periphery of the cornea before lens wear. Bacteria tested were a strain of Staphylococcus aureus or a strain of Staphylococcus epidermidis isolated from human CLPUs. Rabbits were also challenged with S. aureus in the presence of an epithelial defect and in the absence of a contact lens. Corneas were monitored by slitlamp, histology, and microbial culture after 24 hr. RESULTS: No CLPU-like lesions were detected under the following conditions: corneal scratch plus lens wear with no bacteria; corneal scratch plus S. epidermidis colonized lenses; corneal scratch without lens wear and with S. aureus applied to eyes; no scratch plus S. aureus colonized lenses; and corneal scratch plus contact lenses colonized by dead S. aureus. CLPU-like lesions were found only when the corneas were scratched and contact lenses colonized by viable S. aureus were applied to the eye. The histology of the lesions demonstrated a frank epithelial break with underlying stromal infiltration. Only low numbers of bacteria could be cultivated from the corneas with CLPU-like lesions. The CLPU-like lesions shared many similarities with CLPU in humans. CONCLUSION: CLPU-like lesions were only produced by S. aureus and not S. epidermidis in the presence of an epithelial abrasion and contact lens. Thus, we hypothesize that to reduce the incidence of CLPU, contact lenses designed to reduce corneal interaction and repel microbial colonization should be produced.

Animals↗

The influence of Rhizobium and arbuscular mycorrhizal fungi on nitrogen and phosphorus accumulation by Vicia faba.

BACKGROUND AND AIMS: The aim of this study was to investigate the effects of the interactions between the microbial symbionts, Rhizobium and arbuscular mycorrhizal fungi (AMF) on N and P accumulation by broad bean (Vicia faba) and how increased N and P content influence biomass production, leaf area and net photosynthetic rate. METHODS: A multi-factorial experiment consisting of four different legume-microbial symbiotic associations and two nitrogen treatments was used to investigate the influence of the different microbial symbiotic associations on P accumulation, total N accumulation, biomass, leaf area and net photosynthesis in broad bean grown under low P conditions. KEY RESULTS: AMF promoted biomass production and photosynthetic rates by increasing the ratio of P to N accumulation. An increase in P was consistently associated with an increase in N accumulation and N productivity, expressed in terms of biomass and leaf area. Photosynthetic N use efficiency, irrespective of the inorganic source of N (e.g. NO3- or N2), was enhanced by increased P supply due to AMF. The presence of Rhizobium resulted in a significant decline in AMF colonization levels irrespective of N supply. Without Rhizobium, AMF colonization levels were higher in low N treatments. Presence or absence of AMF did not have a significant effect on nodule mass but high N with or without AMF led to a significant decline in nodule biomass. Plants with the Rhizobium and AMF symbiotic associations had higher photosynthetic rates per unit leaf area. CONCLUSIONS: The results indicated that the synergistic or additive interactions among the components of the tripartite symbiotic association (Rhizobium-AMF-broad bean) increased plant productivity.

Biomass↗

Fourier transform-infrared spectroscopic methods for microbial ecology: analysis of bacteria, bacteria-polymer mixtures and biofilms.

Fourier transform-infrared (FT-IR) spectroscopy has been used to rapidly and nondestructively analyze bacteria, bacteria-polymer mixtures, digester samples and microbial biofilms. Diffuse reflectance FT-IR (DRIFT) analysis of freeze-dried, powdered samples offered a means of obtaining structural information. The bacteria examined were divided into two groups. The first group was characterized by a dominant amide I band and the second group of organisms displayed an additional strong carbonyl stretch at approximately 1740 cm-1. The differences illustrated by the subtraction spectra obtained for microbes of the two groups suggest that FT-IR spectroscopy can be utilized to recognize differences in microbial community structure. Calculation of specific band ratios has enabled the composition of bacteria and extracellular or intracellular storage product polymer mixtures to be determined for bacteria-gum arabic (amide I/carbohydrate C-O approximately 1150 cm-1) and bacteria-poly-beta-hydroxybutyrate (amide I/carbonyl approximately 1740 cm-1). The key band ratios correlate with the compositions of the material and provide useful information for the application of FT-IR spectroscopy to environmental biofilm samples and for distinguishing bacteria grown under differing nutrient conditions. DRIFT spectra have been obtained for biofilms produced by Vibrio natriegens on stainless steel disks. Between 48 and 144 h, an increase in bands at approximately 1440 and 1090 cm-1 was seen in FT-IR spectra of the V. natriegens biofilm. DRIFT spectra of mixed culture effluents of anaerobic digesters show differences induced by shifts in input feedstocks. The use of flow-through attenuated total reflectance has permitted in situ real-time changes in biofilm formation to be monitored and provides a powerful tool for understanding the interactions within adherent microbial consortia.

Bacterial Physiological Phenomena↗

Upflow anaerobic sludge blanket reactor--a review.

Biological treatment of wastewater basically reduces the pollutant concentration through microbial coagulation and removal of non-settleable organic colloidal solids. Organic matter is biologically stabilized so that no further oxygen demand is exerted by it. The biological treatment requires contact of the biomass with the substrate. Various advances and improvements in anaerobic reactors to achieve variations in contact time and method of contact have resulted in development of in suspended growth systems, attached growth or fixed film systems or combinations thereof. Although anaerobic systems for waste treatment have been used since late 19th century, they were considered to have limited treatment efficiencies and were too slow to serve the needs of a quickly expanding wastewater volume, especially in industrialized and densely populated areas. At present aerobic treatment is the most commonly used process to reduce the organic pollution level of both domestic and industrial wastewaters. Aerobic techniques, such as activated sludge process, trickling filters, oxidation ponds and aerated lagoons, with more or less intense mixing devices, have been successfully installed for domestic wastewater as well as industrial wastewater treatment. Anaerobic digestion systems have undergone modifications in the last two decades, mainly as a result of the energy crisis. Major developments have been made with regard to anaerobic metabolism, physiological interactions among different microbial species, effects of toxic compounds and biomass accumulation. Recent developments however, have demonstrated that anaerobic processes might be an economically attractive alternative for the treatment of different types of industrial wastewaters and in (semi-) tropical areas also for domestic wastewaters. The anaerobic degradation of complex, particulate organic matter has been described as a multistep process of series and parallel reactions. It involves the decomposition of organic and inorganic matter in the absence of molecular oxygen. Complex polymeric materials such as polysaccharides, proteins, and lipids (fat and grease) are first hydrolyzed to soluble products by extracellular enzymes, secreted by microorganisms, so as to facilitate their transport or diffusion across the cell membrane. These relatively simple, soluble compounds are fermented or anaerobically oxidized, further to short-chain fatty acids, alcohols, carbon dioxide, hydrogen, and ammonia. The short-chain fatty acids (other than acetate) are converted to acetate, hydrogen gas, and carbon dioxide. Methanogenesis finally occurs from the reduction of carbon dioxide and acetate by hydrogen. The initial stage of anaerobic degradation, i.e. acid fermentation is essentially a constant BOD stage because the organic molecules are only rearranged. The first stage does not stabilize the organics in the waste. However this step is essential for the initiation of second stage methane fermentation as it converts the organic material to a form, usable by the methane producing bacteria. The second reaction is initiated when anaerobic methane forming bacteria act upon the short chain organic acids produced in the 1st stage. Here these acids undergo methane fermentation with carbon dioxide acting as hydrogen acceptor and getting reduced to methane. The methane formed, being insoluble in water, escapes from the system and can be tapped and used as an energy source. The production and subsequent escape of methane causes the stabilization of the organic material. The methane-producing bacteria consist of several different groups. Each group has the ability to ferment only specific compounds. Therefore, the bacterial consortia in a methane producing system should include a number of different groups. When the rate of bacterial growth is considered, then the retention time of the solids becomes important parameter. The acid fermentation stage is faster as compared to the methane fermentation stage. This means that a sudden increase in the easily degradable organics will result in increased acid production with subsequent accumulation of acids. This inhibits the methanogenesis step. Acclimatization of the microorganisms to a substrate has been reported to take more than five weeks. Sufficiently acclimated bacteria have shown greater stability towards stress-inducing events such as hydraulic overloads, fluctuations in temperature, fluctuations in volatile acid and ammonia concentrations etc. Several environmental factors can affect anaerobic digestion, by altering the parameters such as specific growth rate, decay rate, gas production, substrate utilization, start-up and response to changes in input. It has long been recognized that an anaerobic process is in many ways ideal for wastewater treatment and has following merits: A high degree of waste stabilization A low production of excess A low nutrient requirements No oxygen requirement Production of methane gas Anaerobic microorganisms, especially methanogens have a slow growth rate. At lower HRTs, the possibility of washout of biomass is more prominent. This makes it difficult to maintain the effective number of useful microorganisms in the system. To maintain the population of anaerobes, large reactor volumes or higher HRTs are required. This may ultimately provide longer SRTs upto 20 days for high rate systems. Thus, provision of larger reactor volumes or higher HRTs ultimately lead to higher capital cost. Among notable disadvantages, it has low synthesis/reaction rate hence long start up periods and difficulty in recovery from upset conditions. Special attention is, therefore, warranted towards, controlling the factors that affect process adversely; important among them being environmental factors such as temperature, pH and concentration of toxic substances. The conventional anaerobic treatment process consists of a reactor containing waste and biological solids (bacteria) responsible for the digestion process. Concentrated waste (usually sewage sludge) can be added continuously or periodically (semi-batch operation), where it is mixed with the contents of the reactor. Theoretically, the conventional digester is operated as a once-through, completely mixed, reactor. In this particular mode of operation the hydraulic retention time (HRT) is equal to the solids retention time (SRT). Basically, the required process efficiency is related to the sludge retention time (SRT), and hence longer SRT provided, results in satisfactory population (by reproduction) for further waste stabilization. By reducing the hydraulic retention time (HRT) in the conventional mode reactor, the quantity of biological solids within the reactor is also decreased as the solids escape with the effluent. The limiting HRT is reached when the bacteria are removed from the reactor faster than they can grow. Methanogenic bacteria are slow growers and are considered the rate-limiting component in the anaerobic digestion process. The first anaerobic process developed, which separated the SRT from the HRT was the anaerobic contact process. In 1963, Young and McCarty (1968) began work, which eventually led to the development of the anaerobic upflow filter (AF) process. The anaerobic filter represented a significant advance in anaerobic waste treatment, since the filter can trap and maintain a high concentration of biological solids. By trapping these solids, long SRT's could be obtained at large waste flows, necessary to anaerobically treat low strength wastes at nominal temperatures economically. Another anaerobic process which relies on the development of biomass on the surfaces of a media is an expanded bed upflow reactor. The primary concept of the process consists of passing wastewater up through a bed of inert sand sized particles at sufficient velocities to fluidize and partially expand the sand bed. One of the more interesting new processes is the upflow anaerobic sludge blanket process (UASB), which was developed by Lettinga and his co-workers in Holland in the early 1970's. The key to the process was the discovery that anaerobic sludge inherently has superior flocculation and settling characteristics, provided the physical and chemical conditions for sludge flocculation are favorable. When these conditions are met, a high solids retention time (at high HRT loadings) can be achieved, with separation of the gas from the sludge solids. The UASB reactor is one of the reactor types with high loading capacity. It differs from other processes by the simplicity of its design. UASB process is a combination of physical & biological processes. The main feature of physical process is separation of solids and gases from the liquid and that of biological process is degradation of decomposable organic matter under anaerobic conditions. No separate settler with sludge return pump is required, as in the anaerobic contact process. There is no loss of reactor volume through filter or carrier material, as in the case with the anaerobic filter and fixed film reactor types, and there is no need for high rate effluent recirculation and concomitant pumping energy, as in the case with fluidized bed reactor. Anaerobic sludge inherently possesses good settling properties, provided the sludge is not exposed to heavy mechanical agitation. For this reason mechanical mixing is generally omitted in UASB-reactors. At high organic loading rates, the biogas production guarantees sufficient contact between substrate and biomass. Regarding the dynamic behaviour of the water phase UASB reactor approaches the completely mixed reactor. For achieving the required sufficient contact between sludge and wastewater, the UASB-system relies on the agitation brought about by the natural gas production and on an even feed inlet distribution at the bottom of the reactor. (ABSTRACT TRUNCATED)

Bacteria, Anaerobic↗

Cell surface characterization of Yarrowia lipolytica IMUFRJ 50682.

In the present work, the surface characteristics of a wild-type strain of Yarrowia lipolytica (IMUFRJ50682) were investigated. Six different methods to characterize cell surfaces--adhesion to polystyrene; hydrophobic interaction chromatography (HIC); microbial adhesion to solvents (MATS) test; zeta potential; microbial adhesion to hydrocarbons (MATH) test; and contact angle measurement (CAM)--were employed to explain the cell surface behaviour of Y. lipolytica (IMUFRJ50682). This Y. lipolytica strain presents significant differences at the cell surface compared with another Y. lipolytica strain (W29) previously reported in the literature. The main difference is related to the higher cell adhesion to non-polar solvents. The proteins present on the cell wall of Y. lipolytica IMUFRJ50682 seem to play an important role in these particular surface characteristics because of the consistent reduction of this yeast hydrophobic character after the action of pronase on its cell wall.

Alkanes↗

Genome-wide characterization of hypothiocyanite stress response in Escherichia coli.

UNLABELLED: Oxidative stress is one of the major methods of microbial population control and pathogen clearing by the mammalian immune system. The methods by which bacteria are able to escape damage by host-derived oxidants such as hydrogen peroxide (H2O2) and hypochlorous acid (HOCl) have been relatively well described, while other oxidants' effects on bacteria and their genetic responses are not as well understood. Hypothiocyanite/hypothiocyanous acid (OSCN-/HOSCN) is one such oxidative stress agent. In this study, we used RNA-sequencing to characterize the global transcriptional response of Escherichia coli to treatment with HOSCN and the impact of deletions of the HOSCN resistance proteins RclA (HOSCN reductase), RclB, and RclC on that response. The HOSCN response of E. coli was different from the previously characterized responses of E. coli to other oxidants such as H2O2, superoxide, or HOCl and distinct from the reported responses of other bacteria such as Streptococcus pneumoniae and Pseudomonas aeruginosa to HOSCN. Strikingly, deletion of any one of the Rcl proteins had very similar effects on the transcriptional response to HOSCN, indicating that any disruption of HOSCN defense in E. coli results in similar impacts, despite the fact that we do not currently understand the mechanism(s) by which RclB and RclC contribute to that defense. IMPORTANCE: Understanding how bacteria sense and respond to oxidative stress provides insights into how our bodies interact with the microbial population within us. In this study, we have characterized the genetic response of E. coli to the important immune oxidant hypothiocyanite and investigated the role of rclABC genes in that response.

Escherichia coli↗

Evidence against hydrogen-based microbial ecosystems in basalt aquifers

It has been proposed that hydrogen produced from basalt-ground-water interactions may serve as an energy source that supports the existence of microorganisms in the deep subsurface on Earth and possibly on other planets. However, experiments demonstrated that hydrogen is not produced from basalt at an environmentally relevant, alkaline pH. Small amounts of hydrogen were produced at a lower pH in laboratory incubations, but even this hydrogen production was transitory. Furthermore, geochemical considerations suggest that previously reported rates of hydrogen production cannot be sustained over geologically significant time frames. These findings indicate that hydrogen production from basalt-ground-water interactions may not support microbial metabolism in the subsurface.

Journal Article↗

Scanning electron microscopy of isolated epithelium of the murine gastrointestinal tract: morphology of the basal surface and evidence for paracrinelike cells.

By using the method of Bjerknes and Cheng, isolated murine gastrointestinal epithelial sheets were prepared for scanning electron microscopy. Examination of isolated epithelium from fundic stomach revealed numerous branched gastric glands. Parietal cells were easily detected bulging from the basal surface of the glandular epithelium. The basal surface membrane of parietal cells appeared smooth, with only sparse microvilluslike projections, whereas adjacent glandular cells had numerous 1- to 2-micron fingerlike projections which interdigitated laterally with similar processes from adjacent cells. Occasionally, paracrinelike cells having long cytoplasmic processes ranging from 10 to 20 micron in length were observed on the basal epithelial surface of the stomach and the colon, but not the small intestine. In isolated intestinal epithelia, the basal surface of crypt epithelial cells showed extensive cytoplasmic interdigitations, but no distinct morphology permitting recognition of individual cell types. Various stages of intestinal crypt bifurcation were seen. Craterlike spaces in the basal surface of crypt epithelium, presumably due to migrating leukocytes, were also numerous. Examination of the luminal surface of the isolated intestinal epithelium revealed that intimate associations between epithelium and mucosal-associated microorganisms were maintained, thus suggesting that minimal alterations in surface morphology were incurred by epithelial isolation. These observations on epithelial structure suggest that isolated gastrointestinal epithelia may be well suited for physiological studies of epithelial function and interactions with the microbial flora.

Animals↗

Autoreactive mechanisms in infective endocarditis.

Infective endocarditis is not a simple interaction between a microbial agent and a cardiac valve. For the infection of a non-bacterial thrombotic vegetation, predisposition is required which is at least partially operative by a decreased suppressor T cell activity. During infection, peripheral blood natural killer cell activity is decreased, but normalizes under anti-microbial therapy. Non-major histocompatibility complex-restricted lymphocytotoxicity to isolated heart cells can be present in one third of patients. Circulating immune complexes normalize during therapy. They may be the cause of many clinical symptoms of infective endocarditis. Anti-bacterial and also anti-sarcolemmal antibodies which are cross-reactive to the bacterium are secreted in a polyclonal immune response. Anti-sarcolemmal antibodies which are cytolytic in vitro in the presence of complement may partly explain the myocardial factor of heart failure in patients with only marginal valve incompetence due to the endocarditic vegetation.

Animals↗

Adherence of a virulent strain of Listeria monocytogenes to the surface of a hepatocarcinoma cell line via lectin-substrate interaction.

Listeria monocytogenes was examined for the presence of surface carbohydrates to ascertain whether surface sugars, if present, would interact with eucaryotic surface carbohydrate receptors. We found that a virulent, but not two avirulent strains had a surface alpha-D-galactose residue as determined by agglutination with Griffonia simplicifolia (GS-I) and other lectins. The virulent strain bound to a human hepatocarcinoma cell line (HepG2), which has a well characterized receptor for alpha-D-galactose. This interaction could be blocked by pretreatment of the HepG2 cells with either alpha-D-galactose or neuraminidase, the latter of which will render the galactose receptor functionally inactive. We propose that the attachment of the virulent Listeria to eucaryotic cells occurs as a result of the interaction of the microbial alpha-D-galactose with that of the eucaryotic galactose receptor. This surface carbohydrate may provide an explanation for the mechanism of attachment and penetration of virulent Listeria into host cells during infection. As such, this may allow for amplification of pathogenesis through intracellular multiplication in nonprofessional phagocytes prior to macrophage involvement.

Agglutination↗

Nonparasitic Nematoda provide evidence for a linear response of functionally important soil biota to increasing livestock density.

Soil acidity, nutrient availability and livestock density have a major influence on the belowground ecological community. As fast decomposition rates are due mostly to bacterial-based pathways and slower decomposition rates mostly to fungal-based pathways, it is helpful to condense empirical information in the so-called Nematode Channel Ratio (NCR). The NCR is shown to be a good indicator of efficiency in soil decomposition processes. We argue that in intensive agroecosystems, other fungivore members of the decomposer food web may outcompete the hyphal-feeding nematodes. We demonstrate how the NCR can be used to set ecological standards for sustainable use of the soil in agroecosystems. To summarize the interactions between the microbial resources and the decomposer nematofauna according to increasing land management, we propose the use of the fifth percentile as proxy for a sustainable environmental quality of grasslands on sandy soils, and the NCR mean as the upper threshold for low-stocked farms.

Animals↗

Intestinal epithelial defense systems protect against bacterial threats.

Numerous bacterial species inhabit the lumen of the human intestine. The epithelial cells that line the intestinal barrier are in direct contact with many of these species and have developed sophisticated strategies to prevent bacterial invasion of host tissue beyond simply providing a physical blockade. Intestinal epithelial cells (IECs) possess receptors that are capable of recognizing bacterial products, and engagement of these receptors results in the production and secretion of immunoregulatory proteins, such as cytokines and chemokines, which mobilize immune effector cells, including macrophages, dendritic cells, and neutrophils. In addition, IECs can produce various lipid-based eicosanoids that can contribute to the orchestration of the immune response. A better understanding of how the intestinal epithelium interacts with its microbial constituents may provide insight into strategies for treatment of enteric infections and inflammatory bowel disease.

Adaptor Proteins, Signal Transducing↗

Drosophila immunity: a large-scale in vivo RNAi screen identifies five serine proteases required for Toll activation.

Unlike mammalian Toll-like Receptors, the Drosophila Toll receptor does not interact directly with microbial determinants but is rather activated upon binding a cleaved form of the cytokine-like molecule Spatzle (Spz). During the immune response, Spz is thought to be processed by secreted serine proteases (SPs) present in the hemolymph that are activated by the recognition of gram-positive bacteria or fungi . In the present study, we have used an in vivo RNAi strategy to inactivate 75 distinct Drosophila SP genes. We then screened this collection for SPs regulating the activation of the Toll pathway by gram-positive bacteria. Here, we report the identification of five novel SPs that function in an extracellular pathway linking the recognition proteins GNBP1 and PGRP-SA to Spz. Interestingly, four of these genes are also required for Toll activation by fungi, while one is specifically associated with signaling in response to gram-positive bacterial infections. These results demonstrate the existence of a common cascade of SPs upstream of Spz, integrating signals sent by various secreted recognition molecules via more specialized SPs.

Animals↗

Effects of the pathogenic Vibrio tapetis on defence factors of susceptible and non-susceptible bivalve species: I. Haemocyte changes following in vitro challenge.

In microbial infections, the interaction between microorganisms and phagocytic cells is a crucial determinant in the outcome of the disease process. We used flow cytometry to study the in vitro interactions between Vibrio tapetis, the bacterium responsible for Brown Ring Disease (BRD) in the Manila clam Ruditapes philippinarum, and haemocytes from three bivalve species: the Manila clam (susceptible to BRD), the hard clam Mercenaria mercenaria and the eastern oyster Crassostrea virginica (both non-susceptible to BRD). Results demonstrated that V. tapetis cells and extracellular products elicit major changes in the haemocytes of R. philippinarum, including decreased viability and phagocytic activity, and altered size and internal structure. V. tapetis was able to kill haemocytes from M. mercenaria and C. virginica but to a far lesser extent than those of R. philippinarum. These results suggest that disease resistance is not solely dependent on a host activity against the pathogen, but is also a function and magnitude of the injury to the host cell by a given pathogen.

Analysis of Variance↗

Crystal structure of the Drosophila peptidoglycan recognition protein (PGRP)-SA at 1.56 A resolution.

Peptidoglycan recognition proteins (PGRPs) form a recently discovered protein family, which is conserved from insect to mammals and is implicated in the innate immune system by interacting with/or degrading microbial peptidoglycans (PGNs). Drosophila PGRP-SA is a member of this family of pattern recognition receptors and is involved in insect Toll activation. We report here the crystal structure of PGRP-SA at 1.56 A resolution, which represents the first example of a "recognition" PGRP. Comparison with the catalytic Drosophila PGRP-LB reveals an overall structure conservation with an L-shaped hydrophilic groove that is likely the PGN carbohydrate core binding site, but further suggests some possible functional homology between recognition and catalytic PGRPs. Consistent with sequence analysis, PGRP-SA does not contain the canonical zinc-binding residues found in catalytic PGRPs. However, substitution of the zinc-binding cysteine residue by serine, along with an altered coordinating histidine residue, assembles a constellation of residues that resembles a modified catalytic triad. The serine/histidine juxtaposition to a threonine residue and a carbonyl oxygen atom, along with conservation of the catalytic water molecule found in PGRP-LB, tantalizingly suggests some hydrolytic function for this member of receptor PGRPs.

Amino Acid Sequence↗

Yeast interactions and wine flavour.

Wine is the product of complex interactions between fungi, yeasts and bacteria that commence in the vineyard and continue throughout the fermentation process until packaging. Although grape cultivar and cultivation provide the foundations of wine flavour, microorganisms, especially yeasts, impact on the subtlety and individuality of the flavour response. Consequently, it is important to identify and understand the ecological interactions that occur between the different microbial groups, species and strains. These interactions encompass yeast-yeast, yeast-filamentous fungi and yeast-bacteria responses. The surface of healthy grapes has a predominance of Aureobasidium pullulans, Metschnikowia, Hanseniaspora (Kloeckera), Cryptococcus and Rhodotorula species depending on stage of maturity. This microflora moderates the growth of spoilage and mycotoxigenic fungi on grapes, the species and strains of yeasts that contribute to alcoholic fermentation, and the bacteria that contribute to malolactic fermentation. Damaged grapes have increased populations of lactic and acetic acid bacteria that impact on yeasts during alcoholic fermentation. Alcoholic fermentation is characterised by the successional growth of various yeast species and strains, where yeast-yeast interactions determine the ecology. Through yeast-bacterial interactions, this ecology can determine progression of the malolactic fermentation, and potential growth of spoilage bacteria in the final product. The mechanisms by which one species/strain impacts on another in grape-wine ecosystems include: production of lytic enzymes, ethanol, sulphur dioxide and killer toxin/bacteriocin like peptides; nutrient depletion including removal of oxygen, and production of carbon dioxide; and release of cell autolytic components. Cell-cell communication through quorum sensing molecules needs investigation.

Bacteria↗