Melding the Old with the New: Trends in Methods Used to Identify, Monitor, and Control Microorganisms on Cultural Heritage Materials.

Microbial activity has an important impact on the maintenance of cultural heritage materials, owing to the key role of microorganisms in many deterioration processes. In order to minimize such deleterious effects, there is a need to fine-tune methods that detect and characterize microorganisms. Trends in microbiology indicate that this need can be met by incorporating modern techniques. All of the methods considered in this review paper are employed in the identification, surveillance, and control of microorganisms, and they have two points in common: They are currently used in microbial ecology (only literature from 2009 to 2015 is included), and they are often applied in the cultural heritage sector. More than 75 peer-reviewed journal articles addressing three different approaches were considered: molecular, sensory and morphological, and biocontrol methods. The goal of this review is to highlight the usefulness of the traditional as well as the modern methods. The general theme in the literature cited suggests using an integrated approach.

Effects of a vinegar-based multi-micronutrient supplement in rats: a multi-pronged assessment of dietary impact.

We determined the effects of continuous access to drinking of water with a vinegar-based multi-micronutrient (VMm) supplement containing rice and fruit vinegars, vitamins, organic acids and sugars during gestation, lactation, and early adulthood in rats. Pregnant rats were provided with reverse-osmosis water or VMm water from the start of pregnancy through the time of weaning. Weaned pups consumed the same drinking water for 3-12 additional weeks. We examined fecal metabolite and microbial profiles, and other physiological parameters. Body weights were less in rats that drank VMm water. Thirty fecal metabolites involved in amino acid and dipeptide metabolism were significantly altered in VMm-supplemented rats. Analysis of microbial 16S rRNA showed enrichment of bacteria in the family S24-7 in VMm-supplemented rats, and one in Ruminococcaceae in controls. Our data show that a VMm-containing beverage can alter growth, and gut metabolism and microbial community. Future work to correlate these parameters is warranted.

Inactivation of foodborne microorganisms using engineered water nanostructures (EWNS).

Foodborne diseases caused by the consumption of food contaminated with pathogenic microorganisms or their toxins have very serious economic and public health consequences. Here, we explored the effectiveness of a recently developed intervention method for inactivation of microorganisms on fresh produce, and food production surfaces. This method utilizes Engineered Water Nanostructures (EWNS) produced by electrospraying of water vapor. EWNS possess unique properties; they are 25 nm in diameter, remain airborne in indoor conditions for hours, contain Reactive Oxygen Species (ROS) and have very strong surface charge (on average 10 e/structure). Here, their efficacy in inactivating representative foodborne bacteria such as Escherichia coli, Salmonella enterica, and Listeria innocua, on stainless steel surfaces and on organic tomatoes, was assessed. The inactivation was facilitated using two different exposure approaches in order to optimize the delivery of EWNS to bacteria: (1) EWNS were delivered on the surfaces by diffusion and (2) a "draw through" Electrostatic Precipitator Exposure System (EPES) was developed and characterized for EWNS delivery to surfaces. Using the diffusion approach and an EWNS concentration of 24,000 #/cm3, the bacterial concentrations on the surfaces were reduced, depending on the bacterium and the surface type, by values ranging between 0.7 to 1.8 logs. Using the EPES approach and for an aerosol concentration of 50,000 #/cm3 at 90 min of exposure, results show a 1.4 log reduction for E. coli on organic tomato surfaces, as compared to the control (same conditions in regards to temperature and Relative Humidity). Furthermore, for L. innocua, the dose-response relationship was demonstrated and found to be a 0.7 and 1.2 logs removal at 12,000 and 23,000 #/cm3, respectively. The results presented here indicate that this novel, chemical-free, and environmentally friendly intervention method holds potential for development and application in the food industry, as a "green" alternative to existing disinfection methods.

Pyomelanin production in Penicillium chrysogenum is stimulated by L-tyrosine.

From a tomb in Upper Egypt we isolated a strain of Penicillium chrysogenum that was capable of producing brown pigment in vitro when grown in a minimal salts medium containing tyrosine. We present evidence that this pigment is a pyomelanin, a compound that is known to assist in the survival of some micro-organisms in adverse environments. We tested type strains of Pe. chrysogenum, which were also able to produce this pigment under similar conditions. Inhibitors of the DHN and DOPA melanin pathways were unable to inhibit the formation of the pigment. Fourier transform IR analysis indicated that this brown pigment is similar to pyomelanin. Pyrolysis-GC/MS revealed the presence of phenolic compounds. Using LC/MS, homogentisic acid, the monomeric precursor of pyomelanin, was detected in supernatants of Pe. chrysogenum cultures growing in tyrosine medium but not in cultures lacking tyrosine. Partial regions of the genes encoding two enzymes in the homogentisic acid pathway of tyrosine degradation were amplified. Data from reverse-transcription PCR demonstrated that hmgA transcription was increased in cultures grown in tyrosine medium, suggesting that tyrosine induced the transcription.

Gut microbiota of an invasive subcortical beetle, Agrilus planipennis Fairmaire, across various life stages.

We characterized gut microbial communities in the emerald ash borer, Agrilus planipennis Fairmaire, an invasive phloem-feeding and wood-boring beetle that has caused extensive mortality to urban and forest ash trees. Analyses included both 16S rRNA gene-based and culture-based approaches. We estimated that the emerald ash borer gut harbors 44, 71, and 49 operational taxonomic units (OTUs(0.03)) in the larval, prepupal, and adult stages, respectively, and a total of 132 OTUs(0.03) when data from the three stages are pooled. The larval gut community shared all its OTUs(0.03) with either the adult or the prepupal gut community, and the adult and prepupal gut communities shared 27 OTUs(0.03). Twenty-two OTUs(0.03) were shared among the three life stages. Rarefaction analyses suggest that these gut microbial communities are close to being completely sampled at the phylum level. Culture-independent techniques yielded a higher diversity of bacteria than did culturing. Three species of bacteria inhabiting guts of emerald ash borer showed cellulolytic activity. The diverse, dynamic, and presumably multifunctional microbial community associated with emerald ash borer guts suggests that invasive insects should be viewed as multispecies complexes and that such an interpretation can improve our ability to develop more effective management approaches.

Indole-3-acetic Acid-producing bacteria are associated with cranberry stem gall.

ABSTRACT Cranberry stem gall is characterized by tumors that girdle stems, thereby killing all distal leaves, flowers, and fruit. Among bacteria isolated from galls, all 11 isolates that were identified as members of the family Enterobacteriaceae caused galls on 50 to 100% of micropropagated cranberry plants that were inoculated. Four of fifteen isolates identified as Pseudomonas spp. caused galls on 10 to 83% of plants inoculated. Twelve of fifteen isolates identified as either Agrobacterium spp. or Rhizobium spp. caused galls on 10 to 50% of plants inoculated, but the galls were smaller than those caused by members of the family Enterobacteriaceae or Pseudomonas spp. There was a positive correlation between the ability of bacteria to produce IAA in vitro and cause galls. In 2002 and 2003, bacteria were isolated from plant and soil samples collected from beds where stem gall had been observed in the past 2 years and beds where stem gall had never been observed. IAA-producing bacteria were common in all samples, although trends were different across years. The results of this study support the hypothesis that IAA-producing bacteria cause cranberry stem gall and suggest that rather than one bacterial species being the cause, multiple strains of bacteria that produce IAA may be responsible for gall formation.

Anatomy of cranberry stem gall and localization of bacteria in galls.

ABSTRACT Cranberry stem gall is characterized by tumors that girdle stems, thereby killing all distal leaves, flowers, and fruit. Bacteria that produce high levels of the plant growth hormone indole-3-acetic acid (IAA) are associated with and believed to cause cranberry stem gall. The anatomy of naturally occurring galls on woody cranberry plants and galls caused by inoculation of micropropagated cranberry plants with Pantoea agglomerans strain 4/99 was consistent with elevated levels of IAA in plants. Field galls exhibited hypertrophy and hyperplasia of tissue external to the vascular cambium, resulting in extensive stem swelling and splitting of the periderm. Similarly, galls on micropropagated plants contained enlarged cortical parenchyma cells. The current year's xylem vessels in field galls were narrow and dense compared with xylem vessels of healthy stems. Curved xylem elements apparently developed de novo within field galls and galls on inoculated plants. Cavities and fissures in both types of galls contained dense aggregates of bacteria. Treatment of micropropagated plants with synthetic IAA caused hypertrophy of cortical parenchyma and formation of adventitious roots. The results support the hypothesis that IAA-producing bacteria cause cranberry stem gall.