Red Kale (Brassica oleracea L. ssp. acephala L. var. sabellica) Induces Apoptosis in Human Colorectal Cancer Cells In Vitro.

This article presents the results of studies investigating the effect of red kale (Brassica oleracea L. ssp. acephala L. var. sabellica) extract on cancer cells (HT-29). The cytotoxicity of the red kale extract was assessed using MTT and LDH assays, while qRT-PCR was employed to analyze the expression of genes associated with the p53 signaling pathway to elucidate the effect of the extract on cancer cells. Furthermore, HPLC-ESI-QTOF-MS/MS was applied to identify bioactive compounds present in red kale. The obtained results indicated that red kale extract reduced the viability and suppressed the proliferation of HT-29 cells (the IC50 value of 60.8 µg/mL). Additionally, mRNA expression analysis revealed significant upregulation of several genes, i.e., casp9, mapk10, mapk11, fas, kat2 b, and ubd, suggesting the induction of cell apoptosis through the caspase-dependent pathway. Interestingly, the study revealed a decrease in the expression of genes including cdk2 and cdk4 encoding cell cycle-related proteins, which may lead to cell cycle arrest. Furthermore, the study identified certain bioactive compounds, such as sinigrin, spirostanol, hesperetin and usambarensine, which could potentially contribute to the apoptotic effect of red kale extracts. However, further investigations are necessary to elucidate the specific role of these individual compounds in the anti-cancer process.

Pilot study on the use of cold atmospheric plasma for preservation of bread.

Cold atmospheric plasma (CAP) is used as an emerging technology for food preservation. In this study, CAP treatment has been applied to bakery products for the first time. The aim of the work was to investigate the effect of the use of CAP on the amount of microorganisms during bread storage. Basic physicochemical properties and bread texture were determined during storage for 0, 3, and 6 days. The study material included gluten-free and mixed wheat-rye bread treated with CAP for 2 and 10 min. The results showed that no mesophilic bacteria or fungi were found after ten minutes of the bread exposure to CAP. In addition, only 2-min non-thermal sterilization resulted in complete inhibition of yeast and mould growth in the gluten-free and wheat-rye bread. A decrease in the microbial growth in the bread was noted; however, a simultaneous decrease in the moisture content of the bread was observed. After the application of plasma for 2 or 10 min, both the gluten-free and mixed wheat-rye bread was characterized by reduced humidity, which also resulted in a significant increase in the hardness and a slight increase in the springiness of the bread. The use of CAP in storage of bread is promising; nevertheless, it is necessary to further study the effect of this treatment in bread with improvers, especially with hydrocolloids and fibers.

Comparison of the Effect of Cold Plasma with Conventional Preservation Methods on Red Wine Quality Using Chemometrics Analysis.

In this study, the effect of cold plasma (CP) on the physicochemical and biological properties of red wine was investigated in comparison with the effects of the conventional preservation method and the combined method. In addition, the effect of storage time after the application of each of the analyzed methods was evaluated. The study examined the effects of the different preservation methods on the pH, color, phenolic content, antioxidant activity and microbiological purity of the red wine. Chemometric analysis was used to discover the relationship between the preservation method used and wine quality. In the wine samples tested, a reduction in phenolic compounds and a decrease in antioxidant activity were noted after storage. This effect was mildest for preservation methods with the addition of potassium metabisulphite and those in which a mixture of helium and nitrogen was used as the working gas. On a positive note, the CP treatment did not affect the color of the wine in a way perceptible to the consumer: ∆E*-1.12 (He/N2; 5 min). In addition, the lowest growth of microorganisms was detected in the CP-treated samples. This indicates the potential of cold plasma as an alternative method to the use of potassium metabisulfite in wine production, which may contribute to its wider use in the alcohol industry in the future.

The impact of cold plasma on the phenolic composition and biogenic amine content of red wine.

The effect of cold plasma (CP) on phenolic compound (PC) and biogenic amine (BA) contents of red wine was investigated for the first time. The influence of CP was compared with the effects of a wine preservation using potassium metabisulfite and a combined method. The PC profile was determined by UPLC-PDA-MS/MS while BAs using DLLME-GC-MS. Chemometric analysis also was used. The content of PCs was 3.1% higher in the sample preserved by CP treatment (5 min, helium/nitrogen) compared to a sample preserved by the addition of potassium metabisulfite (100 mg/L). On a positive note, CP treatment reduced the concentration of BAs in the wine samples. The lowest BA contents were recorded after 10 min of cold plasma (helium/oxygen) treatment with the addition of potassium metabisulfite (1120.85 µg/L). The results may promote interest in CP as a potential alternative method for the preservation of wine and other alcoholic beverages.

Biological Activity of an Epilobium angustifolium L. (Fireweed) Infusion after In Vitro Digestion.

The biological activity of an in vitro digested infusion of Epilobium angustifolium (fireweed) was examined in a model system of intestinal epithelial and colon cancer tissues. The content of selected phenolic compounds in the digested aqueous extract of fireweed was determined using HPLC-ESI-QTOF-MS/MS. Biological activity was examined using the human colon adenocarcinoma cell lines HT-29 and CaCo-2 and the human colon epithelial cell line CCD 841 CoTr. Cytotoxicity was assessed by an MTT assay, a Neutral Red uptake assay, May-Grünwald-Giemsa staining, and a label-free Electric Cell-Substrate Impedance Sensing cytotoxicity assay. The effect of the infusion on the growth of selected intestinal bacteria was also examined. The extract inhibited the growth of intestinal cancer cells HT-29. This effect can be attributed to the activity of quercetin and kaempferol, which were the most abundant phenolic compounds found in the extract after in vitro digestion. The cytotoxicity of the fireweed infusion was dose-dependent. The highest decrease in proliferation (by almost 80%) compared to the control was observed in HT-29 line treated with the extract at a concentration of 250 µg/mL. The fireweed infusion did not affect the growth of beneficial intestinal bacteria, but it did significantly inhibit E. coli. The cytotoxic effect of the fireweed extract indicates that it does not lose its biological activity after in vitro digestion. It can be concluded that the fireweed infusion has the potential to be used as a supporting agent in colon cancer therapy.

Potential Biological Activities of Peptides Generated during Casein Proteolysis by Curly Kale (Brassica oleracea L. var. sabellica L.) Leaf Extract: An In Silico Preliminary Study.

This study is a brief report on the proteolytic activity of curly kale leaf extract against casein. Casein degradation products and an in silico analysis of the biological activity of the peptides obtained was performed. The efficiency of casein hydrolysis by curly kale extract was determined using SDS-PAGE and by peptide concentration determination. The pattern of the enzymatic activity was determined by MALDI-TOF MS analysis. The results showed that α- and ß-casein were more resistant to curly kale extract hydrolysis, whereas κ-casein was absent in the protein profile after 8 h of proteolysis, and all casein fractions were completely hydrolyzed after 24 h of incubation. Based on sequence analysis, seven peptides were identified, with molecular mass in the range of 1151-3024 Da. All the peptides were products of ß-casein hydrolysis. The identified amino acid sequences were analyzed in BIOPEP, MBPDB, and FeptideDB databases in order to detect the potential activities of the peptides. In silico analysis suggests that the ß-casein-derived peptides possess sequences of peptides with ACE inhibitory, antioxidant, dipeptidyl peptidase IV inhibitory, antithrombotic, immunomodulatory, and antiamnesic bioactivity. Our study was first to evaluate the possibility of applying curly kale leaf extract to generate biopeptides through ß-casein hydrolysis.

New Insight into Bacterial Interaction with the Matrix of Plant-Based Fermented Foods.

Microorganisms have been harnessed to process raw plants into fermented foods. The adaptation to a variety of plant environments has resulted in a nearly inseparable association between the bacterial species and the plant with a characteristic chemical profile. Lactic acid bacteria, which are known for their ability to adapt to nutrient-rich niches, have altered their genomes to dominate specific habitats through gene loss or gain. Molecular biology approaches provide a deep insight into the evolutionary process in many bacteria and their adaptation to colonize the plant matrix. Knowledge of the adaptive characteristics of microorganisms facilitates an efficient use thereof in fermentation to achieve desired final product properties. With their ability to acidify the environment and degrade plant compounds enzymatically, bacteria can modify the textural and organoleptic properties of the product and increase the bioavailability of plant matrix components. This article describes selected microorganisms and their competitive survival and adaptation in fermented fruit and vegetable environments. Beneficial changes in the plant matrix caused by microbial activity and their beneficial potential for human health are discussed as well.

Genome and Pangenome Analysis of Lactobacillus hilgardii FLUB-A New Strain Isolated from Mead.

The production of mead holds great value for the Polish liquor industry, which is why the bacterium that spoils mead has become an object of concern and scientific interest. This article describes, for the first time, Lactobacillus hilgardii FLUB newly isolated from mead, as a mead spoilage bacteria. Whole genome sequencing of L. hilgardii FLUB revealed a 3 Mbp chromosome and five plasmids, which is the largest reported genome of this species. An extensive phylogenetic analysis and digital DNA-DNA hybridization confirmed the membership of the strain in the L. hilgardii species. The genome of L. hilgardii FLUB encodes 3043 genes, 2871 of which are protein coding sequences, 79 code for RNA, and 93 are pseudogenes. L. hilgardii FLUB possesses three clustered regularly interspaced short palindromic repeats (CRISPR), eight genomic islands (44,155 bp to 6345 bp), and three (two intact and one incomplete) prophage regions. For the first time, the characteristics of the genome of this species were described and a pangenomic analysis was performed. The concept of the pangenome was used not only to establish the genetic repertoire of this species, but primarily to highlight the unique characteristics of L. hilgardii FLUB. The core of the genome of L. hilgardii is centered around genes related to the storage and processing of genetic information, as well as to carbohydrate and amino acid metabolism. Strains with such a genetic constitution can effectively adapt to environmental changes. L. hilgardii FLUB is distinguished by an extensive cluster of metabolic genes, arsenic detoxification genes, and unique surface layer proteins. Variants of MRS broth with ethanol (10-20%), glucose (2-25%), and fructose (2-24%) were prepared to test the strain's growth preferences using Bioscreen C and the PYTHON script. L. hilgardii FLUB was found to be more resistant than a reference strain to high concentrations of alcohol (18%) and sugars (25%). It exhibited greater preference for fructose than glucose, which suggests it has a fructophilic nature. Comparative genomic analysis supported by experimental research imitating the conditions of alcoholic beverages confirmed the niche specialization of L. hilgardii FLUB to the mead environment.

Morphological and physiological changes in Lentilactobacillus hilgardii cells after cold plasma treatment.

Atmospheric cold plasma (ACP) inactivation of Lentilactobacillus hilgardii was investigated. Bacteria were exposed to ACP dielectric barrier discharge with helium and oxygen as working gases for 5, 10, and 15 min. The innovative approach in our work for evaluation of bacterial survival was the use in addition to the classical plate culture method also flow cytometry which allowed the cells to be sorted and revealed different physiological states after the plasma treatment. Results showed total inhibition of bacterial growth after 10-min of ACP exposure. However, the analysis of flow cytometry demonstrated the presence of 14.4% of active cells 77.5% of cells in the mid-active state and 8.1% of dead cells after 10 min. In addition, some of the cells in the mid-active state showed the ability to grow again on culture medium, thus confirming the hypothesis of induction of VBNC state in L .hilgardii cells by cold plasma. In turn, atomic force microscopy (AFM) which was used to study morphological changes in L. hilgardii after plasma treatment at particular physiological states (active, mid-active, dead), showed that the surface roughness of the mid-active cell (2.70 ± 0.75 nm) was similar to that of the control sample (2.04 ± 0.55 nm). The lack of considerable changes on the cell surface additionally explains the effective cell resuscitation. To the best of our knowledge, AFM was used for the first time in this work to analyze cells which have been sorted into subpopulations after cold plasma treatment and this is the first work indicating the induction of VBNC state in L. hilgardii cells after exposure to cold plasma.

Utilization of brewery wastes in food industry.

Beer is the most popular low-alcohol beverage consumed in large amounts in many countries each year. The brewing industry is an important global business with huge annual revenues. It is profitable and important for the economies of many countries around the world. The brewing process involves several steps, which lead to fermentation of sugars contained in malt and conversion thereof into alcohol and carbon dioxide by yeasts. Beer brewing generates substantial amounts of by-products. The three main brewing industry wastes include brewer's spent grain, hot trub, and residual brewer's yeast. Proper management of these wastes may bring economical benefits and help to protect the environment from pollution caused by their excessive accumulation. The disposal of these wastes is cumbersome for the producers, however they are suitable for reuse in the food industry. Given their composition, they can serve as a low-cost and highly nutritional source of feed and food additives. They also have a potential to be a cheap material for extraction of compounds valuable for the food industry and a component of media used in biotechnological processes aimed at production of compounds and enzymes relevant for the food industry.

Study on Biological Activity of Bread Enriched with Natural Polyphenols in Terms of Growth Inhibition of Tumor Intestine Cells.

A complex plant polyphenolic preparation (PP) was produced from chokeberry, raspberry, wild strawberry, peach, bilberry, apricot, cranberry, and parsley, using ultrafiltration and C18 preparative chromatography. Thirty main compounds were identified in PP (LC-MS), with the highest contribution of cyanidin 3-O-glucoside, p-coumaroyl glucoside, chlorogenic acid, neochlorogenic acid, and isoquercetin. PP was used (at 0.16% m/m) for the production of a sourdough bread (based on rye flour, water, and salt), followed by in vitro digestion. Fluid obtained after PP-enriched bread digestion (EBD fluid) was tested in terms of cytotoxicity, growth inhibition, antioxidant activity, and morphological changes in cancerous intestinal epithelial cells (HT-29) and normal (CCD 841 CoTr). Results show that EBD fluid concentration over 125 µg/mL significantly decreased activity of succinate dehydrogenase in HT-29 cells and reduced their viability of 25%. At this concentration of EBD fluid, modification in cellular morphology was also observed. DPPH analysis revealed that the highest antioxidant activity was observed at concentration of 75 µg/mL, both PP and EBD fluid. Our results show that an introduction of PP into relatively low-polyphenolic, baking products should be carefully considered because polyphenols still retain its biological activity. Antioxidant activity of polyphenols is one of the mechanisms that explains the observed effect of inhibiting the growth of colon cancer cells.

The State of Research on Antimicrobial Activity of Cold Plasma.

Microbiological contamination is a big challenge to the food industry, medicine, agriculture, and environmental protection. For this reason, scientists are constantly looking for alternative methods of decontamination, which ensure the effective elimination of unwanted biological agents. Cold plasma is a new technology, which due to its unique physical and chemical properties becomes a point of interest to a growing group of researchers. The previously conducted experiments confirm its effective action, e.g. in the disinfection of skin wounds, air, and sewage treatment, as well as in food preservation and decontamination. The reactive compounds present in the plasma: high-energy electrons, ionized atoms and molecules, and UV photons are the key factors that cause an effective reduction in the number of microorganisms. The mechanism and effectiveness of the cold plasma are complex and depend on the process parameters, environmental factors and the type and properties of the microorganisms that are to be killed. This review describes the current state of knowledge regarding the effectiveness of the cold plasma and characterizes its interaction with various groups of microorganisms based on the available literature data.Microbiological contamination is a big challenge to the food industry, medicine, agriculture, and environmental protection. For this reason, scientists are constantly looking for alternative methods of decontamination, which ensure the effective elimination of unwanted biological agents. Cold plasma is a new technology, which due to its unique physical and chemical properties becomes a point of interest to a growing group of researchers. The previously conducted experiments confirm its effective action, e.g. in the disinfection of skin wounds, air, and sewage treatment, as well as in food preservation and decontamination. The reactive compounds present in the plasma: high-energy electrons, ionized atoms and molecules, and UV photons are the key factors that cause an effective reduction in the number of microorganisms. The mechanism and effectiveness of the cold plasma are complex and depend on the process parameters, environmental factors and the type and properties of the microorganisms that are to be killed. This review describes the current state of knowledge regarding the effectiveness of the cold plasma and characterizes its interaction with various groups of microorganisms based on the available literature data.

Isolation and characterization of a new fructophilic Lactobacillus plantarum FPL strain from honeydew.

In the present study, a Lactobacillus plantarum FPL strain exhibiting fructophilic behavior has been isolated for the first time from honeydew. It is a probably syntrophic bacterium inhabiting the gastrointestinal tract of Coccus hesperidum L. and taking part in sugar metabolism. The promising growth characteristics and biochemical properties of Lb. plantarum FPL indicate that this may be a facultatively fructophilic species, whose properties are not associated with the loss of the alcohol/acetaldehyde dehydrogenase gene. The article attempts to classify the peculiar behavior of this strain by means of tests that are characteristic for FLAB as well as through a classic identification approach. In this study, we used a reference strain Lb. plantarum NRRL B-4496, which showed no fructophilic properties. With the FLAB group, the new strain shares the habit, such as a fructose-rich environment, the preference of this sugar for growth, and similar growth curves. However, it exceeds FLAB in terms of osmotolerance to high sugar content. The fructophilic Lb. plantarum FPL strain can proliferate and grow on a medium wherein the sugar concentration is 45 and 50% (w/v). Our findings indicate that honeydew can be a promising source of new fructophilic lactic acid bacteria.

Composition of lactic acid bacteria during spontaneous curly kale (Brassica oleracea var. sabellica) fermentation.

The present work is the first report on spontaneous fermentation of curly kale and characteristics of autochthonous lactic acid bacteria (LAB). Our results indicate that curly kale fermentation is the new possibility of the technological use of this vegetable. Bacteria representing ten different species were isolated from three phases of curly kale fermentation and identified by MALDI-TOF mass spectrometry and 16S rRNA gene sequencing. Among them, four species were identified as Lactobacillus spp. (Lb. plantarum 332, Lb. paraplantarum G2114, Lb. brevis R413, Lb. curvatus 154), two as Weissella spp. (W. hellenica 152, W. cibaria G44), two as Pediococcus spp. (P. pentosaceus 45AN, P. acidilactici 2211), one as Leuconostoc mesenteroides 153, and one as Lactococcus lactis 37BN. The functional properties of isolates, i.e. acid, NaCl and bile salt tolerance, enzyme activities, adhesion to hydrocarbons, and antibiotic resistance, were examined. Among the tested strains, Lb. plantarum 332, Lb. paraplantarum G2114, P. pentosaceus 2211, and Lb. brevis R413 exhibited the best hydrophobicity value and high tolerance to bile salts, NaCl, and low pH.

Studies on the removal of Cd ions by gastrointestinal lactobacilli.

Accumulation of toxic metal ions in food and water is nowadays a growing health-related problem. One detoxification method involves the use of microorganisms naturally inhabiting the gastrointestinal tract (GIT). The purpose of this study was to prove that lactic acid bacteria derived from the GIT are able to effectively remove Cd2+ from water solution. Seven strains of lactobacilli, out of 11 examined, showed tolerance to high concentrations of cadmium ions. The metal-removal efficiencies of these seven lactobacilli ranged from 6 to 138.4 µg/h mg. Among these bacteria, Lactobacillus gallinarum and Lactobacillus crispatus belonged to the highest (85%) Cd-removal efficiency class. An analysis of the zeta potential (ζ) indicated that the bacterial cell surface had a negative charge at the pH ranging from 3 to 10. The presence of carboxyl, amide, and phosphate groups was favorable for Cd2+ binding to the cell surface, which found confirmation in FTIR-ATR spectra. Elemental SEM/EDS analysis and TEM imaging not only confirmed the adsorption of Cd2+ on the cell envelope but also gave us a reason to suppose that Lb. crispatus accumulates metal ions inside the cell. Our findings open perspectives for further research on the new biological function of GIT lactobacilli as natural biosorbents.

The first report of the physicochemical structure of chitin isolated from Hermetia illucens.

This is the first report on the physicochemical properties of chitin obtained from larvae and imagoes of black soldier flies (Hermetia illucens). Scanning electron microscopy revealed differences in surface morphologies of the two types of chitin. The crystalline index values of chitins from adult flies and larvae were 24.9% and 35%, respectively. This is a trait that differentiates these biopolymers from chitins extracted from other sources described so far. X-ray diffraction patterns and IR spectroscopy revealed that both types of samples of chitin were in an α crystalline form. Also, the results of elemental analysis, thermal stabilities and FTIR spectroscopy of the chitins from larvae and adults of H. illucens were similar, which points to a general similarity in their physicochemical structure.

Physicochemical characterization of exopolysaccharides produced by Lactobacillus rhamnosus on various carbon sources.

The impact of five carbohydrate sources (glucose, maltose, galactose, sucrose, and lactose) on the chemical composition, structure, morphology, and physicochemical properties, as well as, viscosity of exopolysaccharides (EPSs) produced by Lactobacillus rhamnosus E/N was investigated. GLC-MS analysis and 2DNMR spectroscopy showed that the EPSs had the same primary structure independently of the carbon source used in the growth medium. The following EPS composition was elucidated: four rhamnose, two glucose, and one galactose residue with a pyruvate substituent. Molecular masses (M(w)) were determined by gel permeation chromatography, which revealed differences in M(w) distribution. EPS-Gal, EPS-Suc, and EPS-Lac showed heterogenic fractions of a high and low molecular weight, while EPS-Mal and EPS-Glc contained only a high-molecular-weight fraction. AFM microscopy revealed morphological differences in chain length, thickness, and branching. Differences in the Mw ratio and thickness of the polymer chain were correlated with high viscosity of EPS solutions. Our results indicate that a single bacterial strain, depending on the carbon source in the medium, can produce EPSs of different rheological properties.

The effect of moonlighting proteins on the adhesion and aggregation ability of Lactobacillus helveticus.

The goal of this study was to identify moonlighting proteins in Lactobacillus helveticus that play an important role in adhesion and aggregation. The label-free method was used for identification and analysis of expression of cellular proteins. The analysis revealed the presence of eight moonlighting proteins in the cell envelope of Lb. helveticus. The tested strains mainly differed with respect to the presence of S-layer proteins and the level of expression of moonlighting proteins in Lb. helveticus strain T159. These surface proteins give the cell a hydrophobic character and play a role in specific interactions with intestinal epithelium cells and with other bacteria. In Lb. helveticus T159, the S-layer associated with moonlighting proteins could act as adherence factors, which was evidenced by the high capability of adhesion, auto- and coaggregation. The hydrophobicity, adhesion and aggregation abilities provide biological activities in food products and they are regarded as an important criterion for probiotic selection.

Biosorption of Al(+3) and Cd(+2) by an exopolysaccharide from Lactobacillus rhamnosus.

The aim of this study was to assess the removal of Cd(2+) and Al(3+) from aqueous solutions by an exopolysaccharide (EPS) from Lactobacillus rhamnosus E/N. The biosorption kinetics of EPS for Cd(2+) and Al(3+) were studied by equilibrium dialysis after incubation from 0 h to 58 h. The actual concentrations of Cd(2+) and Al(3+) in external solutions were measured by a fluorescence method after complexation with 8-hydroxyquinoline. Binding of metal ions by EPS was observed, with maximum adsorption after 0 to 24 and 48 to 58 h for Cd(2+) , and after 24 h for Al(3+) . The existing analytical method, involving the binding of metal ions was measured using a dialysis membrane was significantly improved in our study. This new method could be applied in future studies of this type. The results of this study demonstrate that EPS from Lb. rhamnosus E/N has a Cd(2+) and Al(3+) biosorption capacity. Surface adsorption of the metal ions at surface of EPS was confirmed through scanning electron microscopy. Fourier transform-infrared spectroscopy analysis suggests a role of the functional groups ─OH, C═O, and COO(-) from EPS in the binding of Cd(2+) and Al(3+) ions.

The effect of cell surface components on adhesion ability of Lactobacillus rhamnosus.

The aim of this study was to analyze the cell envelope components and surface properties of two phenotypes of Lactobacillus rhamnosus isolated from the human gastrointestinal tract. The ability of the bacteria to adhere to human intestinal cells and to aggregate with other bacteria was determined. L. rhamnosus strains E/N and PEN differed with regard to the presence of exopolysaccharides (EPS) and specific surface proteins. Transmission electron microscopy showed differences in the structure of the outer cell surface of the strains tested. Bacterial surface properties were analyzed by Fourier transform infrared spectroscopy, fatty acid methyl esters and hydrophobicity assays. Aggregation capacity and adhesion of the tested strains to the human colon adenocarcinoma cell line HT29 was determined. The results indicated a high adhesion and aggregation ability of L. rhamnosus PEN, which possessed specific surface proteins, had a unique fatty acid content, and did not synthesize EPS. Adherence of L. rhamnosus was dependent on specific interactions and was promoted by surface proteins (42-114 kDa) and specific fatty acids. Polysaccharides likely hindered bacterial adhesion and aggregation by masking protein receptors. This study provides information on the cell envelope constituents of lactobacilli that influence bacterial aggregation and adhesion to intestinal cells. This knowledge will help to understand better their specific contribution in commensal-host interactions and adaptation to this ecological niche.