Low-Resolution Raman Spectroscopy for the detection of contaminant species in algal bioreactors.

Fouling of aquatic systems by harmful microalgal and cyanobacterial species is an environmental and public health concern. Microalgal bioreactors are engineered ecosystems for the cultivation of algal biomass to meet the increasing demand for alternative protein sources and algae-derived products. Such bioreactors are often open or semi-open ponds or raceways that are prone to contamination by contaminant photosynthetic microorganisms, including harmful cyanobacterial species (HCBs). HCBs affect the quality of products through the accumulation of off-flavours, reducing their acceptance by consumers, and through the production of several different toxins collectively known as cyanotoxins. The density of cultured species within the bioreactor environment creates difficulty in detecting low concentrations of contaminant cells, and there is currently no technology enabling rapid monitoring of contaminations. The present study demonstrates the potential of Low-Resolution Raman Spectroscopy (LRRS) as a tool for rapid detection of low concentrations of HCBs within dense populations of the spirulina (Arthrospira platensis) cultures. An LRRS system adapted for the direct measurement of raw biomass samples was used to assemble a database of Raman spectral signatures, from eight algal and cyanobacterial strains. This dataset was used to develop both quantitative and discriminative chemometric models. The results obtained from the chemometric analyses demonstrate the ability of the LRRS to detect and quantify algal and cyanobacterial species at concentrations as low as 103 cells/mL and to robustly discriminate between species at concentrations of 104 cells/mL. The LRRS and chemometric analyses were further able to detect the presence of low concentrations (103cells/mL) of contaminating species, including the toxic cyanobacterium Microcystis aeruginosa, within dense (>107 cells/mL) spirulina cultures. The results presented provide a first demonstration of the potential of LRRS technology for real-time detection of contaminant species within microalgal bioreactors, and possibly for early detection of developing harmful algal blooms in other aquatic ecosystems.

Pomegranate Fruit Growth and Skin Characteristics in Hot and Dry Climate.

Pomegranate (Punica granatum L.) fruit is well known for its health-beneficial metabolites. The pomegranate peel consists of an inner thick spongy white tissue, and an outer smooth skin layer that accumulates anthocyanins in red cultivars when ripe. The skin is made up of epidermis cells covered by a cuticle, the latter being the first target of cracking and russeting. The present study focuses on the effect of Israel's hot and dry climate on pomegranate growth, to elucidate the derived effects on fruit skin characteristics and its putative resistance to the building pressure from fruit expansion. Experiments were conducted for four years, in four orchards located in different regions of the country, each with a different typical microclimate. Fruit-growth parameters were followed using remote-sensing tools, microscopic study, and mineral analysis of the skin, followed by determination of the peel's elastic modulus. Fruit expanded in two phases: a short rapid phase followed by a gradual phase with a sigmoidal growth-rate pattern. Extreme hot and dry climate during the period of maximal growth rate was associated with restricted growth and a high proportion of small-size fruit. Anatomical study indicated that the skin of mature pomegranate fruit is made up of epidermal cells that are relatively flat and spaced apart, and is expected to be less durable against internal pressure. In contrast, skin of early immature fruit has two layers of dense and rounded epidermis, and is expected to be more resistant to cracking. Tensile strength studies confirmed this trend-skin of mature fruit had a lower elastic modulus than young fruit. However, restrained growth due to extreme environmental cues may result in better resistance of the mature pomegranate fruit to cracking, and in better skin quality and appearance, albeit small fruits. On the other hand, temperate climate at the beginning of the growth period, which allows high growth rate and high daily shrinkage, leads to pomegranate skin disorders.

Effects of steam sterilization on reduction of fungal colony forming units, cannabinoids and terpene levels in medical cannabis inflorescences.

Medical cannabis (MC) production is a rapidly expanding industry. Over the past ten years, many additional phytocannabinoids have been discovered and used for different purposes. MC was reported beneficial for the treatment of a variety of clinical conditions such as analgesia, multiple sclerosis, spinal cord injuries, Tourette's syndrome, epilepsy, glaucoma, Parkinson disease and more. Yet, there is still a major lack of research and knowledge related to MC plant diseases, both at the pre- and postharvest stages. Many of the fungi that infect MC, such as Aspergillus and Penicillium spp., are capable of producing mycotoxins that are carcinogenic, or otherwise harmful when consumed, and especially by those patients who suffer from a weakened immune system, causing invasive contamination in humans. Therefore, there are strict limits regarding the permitted levels of fungal colony forming units (CFU) in commercial MC inflorescences. Furthermore, the strict regulation on pesticide appliance application in MC cultivation exacerbates the problem. In order to meet the permitted CFU limit levels, there is a need for pesticide-free postharvest treatments relying on natural non-chemical methods. Thus, a decontamination approach is required that will not damage or significantly alter the chemical composition of the plant product. In this research, a new method for sterilization of MC inflorescences for reduction of fungal contaminantstes was assessed, without affecting the composition of plant secondary metabolites. Inflorescences were exposed to short pulses of steam (10, 15 and 20 s exposure) and CFU levels and plant chemical compositions, pre- and post-treatment, were evaluated. Steam treatments were very effective in reducing fungal colonization to below detection limits. The effect of these treatments on terpene profiles was minor, resulting mainly in the detection of certain terpenes that were not present in the untreated control. Steaming decreased cannabinoid concentrations as the treatment prolonged, although insignificantly. These results indicate that the steam sterilization method at the tested exposure periods was very effective in reducing CFU levels while preserving the initial molecular biochemical composition of the treated inflorescences.

Quantification of bacteria in water using PLS analysis of emission spectra of fluorescence and excitation-emission matrices.

Bacterial contamination of drinking water is a considerable concern for public health. Tryptophan-like fluorescence (TLF) has been widely suggested to enable fast and inexpensive monitoring and quantification of bacterial contamination of water. Typically, TLF is determined at a certain excitation (ex)/emission (em) wavelengths pair. The aim of this study was to assess fluorescence spectroscopy supported with partial least squares (PLS) algorithms as a tool for a rapid evaluation of the microbial quality of water, by comparing the use of a single ex/em wavelengths pair, of the spectrum of emission obtained at a single excitation wavelength to that of whole excitation-emission matrices (EEMs). For that, laboratory-grown Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa were studied as the model systems, as well as 90 groundwater samples from 6 different wells in Israel. The groundwater samples were characterized for fluorescence emission, coliforms, fecal coliforms, fecal streptococci and heterotrophic plate counts. The PLS analysis of emission spectra and, especially, of EEMs was capable of meaningfully reducing the detection limit of microorganisms in model systems, as compared with the single ex/em wavelengths pair-based determination commonly used, reaching a detection threshold as low as 10 CFU/ml. Use of PLS-analyzed EEMs becomes beneficial also in terms of correlation and similarity between the actual and predicted bacterial concentrations. Similarly, improved detection of bacteria was also achieved in groundwater samples. Furthermore, at a level of >104 CFU/ml, use of EEMs coupled with PLS enabled discrimination between E. coli, B. subtilis and P. aeruginosa.