Hyphal growth of Penicillium rubens in changing relative humidity.

When considering mold prevention strategies, the environmental conditions in which fungi grow need to be taken into consideration. This environment is often characterized by a time-dependent relative humidity, and porous substrate. Growth has mainly been investigated in steady-state experiments. Therefore, the goal of this study is to understand the hyphal growth of Penicillium rubens on porous gypsum, under dynamic humidity conditions. Spores of P. rubens were inoculated on porous gypsum containing nutrients, and placed in a small incubation chamber, allowing for microscopic hyphal observation. The relative humidity in this chamber varied multiple times between a high (close to 100%) and low value (35%, 55%, or 75%). The hyphae reacted to a lowered relative humidity by an immediate growth stop and dehydration. When the relative humidity was increased again, the hyphae re-hydrated and three responses were found: regrowing after approximately 4 h, after a time equal to the germination time, or no regrowth at all. No substantial regrowth was found for fluctuations faster than 4 h. This time-scale was found for multiple decreases in relative humidity, and has been reported for the first time. KEY POINTS: • Hyphae restart growth after a characteristic time of approximately 4 h. • Relative humidity fluctuations of 3 h can suppress hyphal growth. • Hyphae do not regrow after a severe desiccation and short periods of high humidity.

Film Formation of High Tg Latex Using Hydroplasticization: Explanations from NMR Relaxometry.

The film formation of acrylic latex dispersions, containing different amounts of carboxylic acid functional groups by the incorporation of methacrylic acid (MAA), was studied with GARField 1H NMR at various relative humidities (RH). Polymer particles with glass-transition temperatures in the range from 26 to 50 °C formed films at room temperature because of hydroplasticization. It was found that with an increased drying rate due to lower RH, the evaporation flux of water was limited by the latex polymer. Only in the second stage of drying this phenomenon was more obvious with increasing MAA content. 1H NMR relaxometry was used to study the change of hydrogen mobilities during film formation and hardening of the films. This showed that the drying rate itself had no impact on the hydrogen mobility in the latex films as measured via the T2 relaxation time. Hydrogen mobilities of water and the mobile polymer phase only significantly decrease after most water has evaporated. This implies that the rigidity of the polymers increases with the evaporation of water that otherwise plasticizes the polymer through hydrogen bonding with the carboxylic acid groups. This hardening of the polymer phase is essential for applications in a coating. The hydrogen mobilities were affected by the MAA concentration. Densities of mobile hydrogens increase with increasing MAA content. This is expected if the mobile protons are contained in the MAA groups. The result thus confirms the role of carboxylic acid groups in hydrogen bonding and plasticization of the copolymers. Hydrogen mobilities, however, decrease with increasing MAA content, which is hypothesized to be caused by the formation of dimers of carboxylic acid groups that still hold water. They still enable short-range polymer hydrogen mobility due to hydroplasticization but limit long-range polymer mobility due to interaction between the carboxylic acid groups.

The Indoor Fungus Cladosporium halotolerans Survives Humidity Dynamics Markedly Better than Aspergillus niger and Penicillium rubens despite Less Growth at Lowered Steady-State Water Activity.

UNLABELLED: Indoor fungi cause damage in houses and are a potential threat to human health. Indoor fungal growth requires water, for which the terms water activity (aw) and relative humidity (RH) are used. The ability of the fungi Aspergillus niger, Cladosporium halotolerans, and Penicillium rubens at different developmental stages to survive changes in aw dynamics was studied. Fungi grown on media with high aw were transferred to a controlled environment with low RH and incubated for 1 week. Growth of all developmental stages was halted during incubation at RHs below 75%, while growth continued at 84% RH. Swollen conidia, germlings, and microcolonies of A. niger and P. rubens could not reinitiate growth when retransferred from an RH below 75% to a medium with high aw All developmental stages of C. halotolerans showed growth after retransfer from 75% RH. Dormant conidia survived retransfer to medium with high aw in all cases. In addition, retransfer from 84% RH to medium with high aw resulted in burst hyphal tips for Aspergillus and Penicillium Cell damage of hyphae of these fungi after incubation at 75% RH was already visible after 2 h, as observed by staining with the fluorescent dye TOTO-1. Thus, C. halotolerans is more resistant to aw dynamics than A. niger and P. rubens, despite its limited growth compared to that of these fungi at a lowered steady-state aw The survival strategy of this phylloplane fungus in response to the dynamics of aw is discussed in relation to its morphology as studied by cryo-scanning electron microscopy (cryo-SEM). IMPORTANCE: Indoor fungi cause structural and cosmetic damage in houses and are a potential threat to human health. Growth depends on water, which is available only at certain periods of the day (e.g., during cooking or showering). Knowing why fungi can or cannot survive indoors is important for finding novel ways of prevention. Until now, the ability of fungi to grow on media with little available water at steady state (unchanging conditions) has been important for evaluating whether a fungus can grow indoors. In the present study, we found that the fungus Cladosporium halotolerans, a common indoor fungus, is more resistant to changes in available water than the fungi Aspergillus niger and Penicillium rubens, despite the fact that the latter fungi can grow on media with low water availability. We concluded that the ability of fungi to deal with changes in humidity is at least as important as the ability to grow on low-water media.

Separate effects of moisture content and water activity on the hyphal extension of Penicillium rubens on porous media.

To prevent indoor fungal growth, understanding the moisture relations of fungi is a key element. Indoor moisture is quantified by the relative humidity (RH). RH controls the water activity of the indoor materials that fungi grow on, a well-studied parameter known to limit fungal growth. RH, however, also controls the amount of water present in these materials, the moisture content. The significance of the moisture content of these materials to indoor fungal growth is currently overlooked. In the work reported here, growth experiments with the indoor fungus Penicillium rubens on gypsum substrates were performed to test whether the moisture content influences growth on porous materials. Second, we report the development of a video microscopy method that for the first time quantified hyphal growth on a porous material. It is found that a higher moisture content leads to earlier colonization and higher hyphal extension rates. This is a fundamental step in unravelling the effect of RH on indoor fungal growth. The real-time monitoring of colonization of gypsum provides a new view of growth on indoor surfaces.

Coating formation during drying of ß-lactoglobulin: gradual and sudden changes.

The drying dynamics of protein coatings is of importance for many applications. The main focus of research so far was to investigate macroscopic properties of protein coatings, leaving drying dynamics virtually unexplored. A unique combination of techniques is used to monitor drying of a coating containing the protein ß-lactoglobulin. The techniques used cover both macroscopic and microscopic aspects of the drying process. For all water fractions amenable to diffusing wave spectroscopy analysis (xw > 0.2 w/w), the tracer particles diffuse in the coating as in a Newtonian viscous medium. Magnetic resonance imaging shows both protein and water are distributed homogeneously over the coating during drying, up to water fractions above 0.2 w/w. When drying continues to lower water fractions, sudden transitions in drying behavior are observed by both dynamic vapor sorption and IR spectroscopy, which we suggest are due to changes in molecular interactions caused by dehydration of the protein backbone.

Penicillium rubens germination on desiccated and nutrient-depleted conditions depends on the water activity during sporogenesis.

Fungal growth often appears in a surrounding where water and nutrients are scarce. The impact of this environment during sporogenesis on subsequent growth is often neglected. This study investigates the effect of water availability during sporogenesis on subsequent early growth. Therefore, a carbon-depleted substrate was constructed. Humidity is then the only parameter of interest. The water conditions during sporogenesis, and during subsequent growth, were varied. This is a stressing environment: no carbon source is present, and water provided solely via the vapour. The lag time, tl, and initial growth rate, µfp, of the germ tubes were monitored. The effect of aw history on germination and initial growth depends on the RH of the environment. Only at low RH do spores produced at low aw have a smaller tl and higher µfp compared to those grown at high aw. This result was remarkably pronounced when the substrate was also made hydrophobic: growth only occurred when spores were developed at low aw and placed in high RH. Spores grown on lowered aw attract more water. It is hypothesized that this attraction affects subsequent growth behaviour, and is the reason why growth on hydrophobic glass only prevails in the condition of high RH and lowered aw history. We demonstrate the influence of cultivation conditions on germination, which becomes more pronounced in a more desiccated environment.

Oil type and cross-linking influence growth of Aureobasidium melanogenum on vegetable oils as a single carbon source.

Aureobasidium melanogenum is the main fungus found in a spontaneously formed biofilm on a oil-treated wood. This dark colored biofilm functions as a protective coating. To better understand biofilm formation, in this study A. melanogenum was cultured on olive oil and raw linseed oil. Metabolic activity and oil conversion were measured. The results show that A. melanogenum is able to grow on linseed oil and olive oil as a single carbon source. The fungus produces the enzyme lipase to convert the oil into fatty acids and glycerol. Metabolic activity and oil conversion were equal on linseed oil and olive oil. The fungus was not able to grow on severe cross-linked linseed oil, meaning that the degree of cross-linking of the oil is important for growth of A. melanogenum. Dark coloring of the colony was seen on linseed oil, which might be a stress response on the presence of autoxidation products in linseed oil. The colony on olive oil showed delayed melanin production indicating an inhibitory effect of olive oil on melanin production.

Conidia of Penicillium rubens formed at low water activities can attract more water.

To address the problem of indoor fungal growth, understanding the influence of moisture conditions on the fungal colonization process is crucial. This paper explores the influence of past moisture conditions on current processes. Specifically, it studies the growth and water sorption of conidia of Penicillium rubens formed at lower water activities (ranging from 0.86 to 0.99). For the first time, dynamic vapor sorption (DVS) is applied as a tool to quantify the water sorption of conidia as a function of the water activity at conidiation. Furthermore, growth experiments on agar and gypsum substrates are reported that relate hyphal growth rates of the mycelium from pretreated conidia to the water activity at conidiation. No effect of the conidiation water activity on mycelial growth rates is found on either gypsum or agar. It is found, however, that conidia formed at lower activities have a higher dry weight and attract more water from humid air. It is shown that both phenomena can be explained by conidia from lower activities carrying higher amounts of compatible solutes, glycerol in particular. The enhanced sorption observed in this study might constitute a mechanism through which solute reserves contribute to survival during the early steps of fungal colonization.

A microscopy study of hyphal growth of Penicillium rubens on gypsum under dynamic humidity conditions.

To remediate indoor fungal growth, understanding the moisture relations of common indoor fungi is crucial. Indoor moisture conditions are commonly quantified by the relative humidity (RH). RH is a major determinant of the availability of water in porous indoor surfaces that fungi grow on. The influence of steady-state RH on growth is well understood. Typically, however, the indoor RH constantly changes so that fungi have to endure frequent periods of alternating low and high RH. Knowledge of how common indoor fungi survive and are affected by the low-RH periods is limited. In particular, the specific effects of a drop in RH on the growth of the mycelium remain unclear. In this work, video microscopy was used to monitor hyphal growth of Penicillium rubens on gypsum substrates under controlled dynamic humidity conditions. The effect of a single period of low RH (RH = 50-90%) interrupting favourable conditions (RH = 97%) was tested. It was found that hyphal tips ceased to extend when exposed to any tested decrease in RH. However, new hyphal growth always emerges, seemingly from the old mycelium, suggesting that this indoor fungus does not rely only on conidia to survive the humidity patterns considered. These findings are a fundamental step in unravelling the effect of RH on indoor fungal growth.

T2 distribution spectra obtained by continuum fitting method using a mixed Gaussian and Exponential kernel function.

Static (1)H NMR Free Induction Decay (FID) signals of polymer solids contain a lot of information about the molecular dynamics. A T2 analysis of the FID has generally been performed in terms of discrete two- or three-component models. However, this requires a priori assumption of the number of proton species before analysis. This paper presents a method of analyzing the FIDs of the polymer solid samples in terms of a continuous T2 distribution. A mixed Gaussian and Exponential kernel function was used to represent the true characteristic of FIDs of the polymer solids. A simple and realistic assumption has been made to reduce the number of degrees of freedom in the continuum fitting and to make the fitting stable. An experimental static (1)H NMR FID of a typical polymer solid sample was analyzed as an example in the end to demonstrate the application of this method.

High spatial resolution NMR imaging of polymer layers on metallic substrates.

High spatial resolution NMR imaging techniques have been developed recently to measure the spatial inhomogeneity of a polymer coating film. However, the substrates of the polymer coatings for such experiments are generally required to be non-metallic, because metals can interact with static magnetic fields B(0) and RF fields B(1) giving rise to artifacts in NMR images. In this paper we present a systematic study on the effects of metallic substrates on 1D profiles obtained by high resolution NMR imaging. The off-resonance effect is discussed in detail in terms of the excitation profile of the RF pulses. We quantitatively show how the NMR signal intensities change with frequency offset at different RF pulse lengths. The complete NMR profiles were simulated using a Finite Element Analysis method by fully considering the inhomogeneities in both B(1) and B(0). The excellent agreement between the calculated and measured NMR profiles on both metallic and non-metallic substrates indicates that the experimental NMR profiles can be reproduced very well by numerical simulations. The metallic substrates can disturb the RF field of the coil by eddy current effect and therefore change the NMR profiles. To quantitatively interpret the NMR profile of a polymer layer on a metallic substrate, the profile has to be divided by the profile of a reference on the same metallic substrate located at the same distance from the coil.