Fruit Pathogens Research Articles

Incidence of Pathogens in Fruits and Vegetables in Tropical Bwari Town

Introduction: In numerous tropical regions of developing nations, the prevalence of foodborne illnesses is perceived to be more severe than in developed countries, due to insufficient food safety initiatives or the lack of a structured institutional framework for street food vendors. The study's objectives of examining microorganisms associated with fruits and vegetables, the prevailing temperature and the hygiene practices of various fruit and vegetable sellers were all met. Methods: A self-designed questionnaire completed by twenty vendors were used. Laboratory techniques like serial dilution, biochemical testing, and culture techniques were used. A handheld thermometer was used to determine the prevailing temperature, and descriptive statistics using Microsoft Excel was used to analyze data. Results and Discussion: Statistical analysis revealed that 80% of vendors had washing facilities, 70% worked in a clean environment, 20% washed their hands before handling fruits, 70% dressed correctly, and 70% kept their fruits and vegetables above the WHO temperature standard. A laboratory investigation revealed the bacteria counts of (watermelon being the highest 8.3×105, cucumber 5.4×105, carrot 2.3×105, orange 3.7×105). Microbial species encountered were Staphylococcus aureus, Escherichia coli, Salmonella species, amongst others, which increases health risk. Conclusion: The study concluded that temperatures, handling processes, and inadequate hygiene all had impact on fruits and vegetables sold in tropical Bwari area. It also established a link between temperature and the prevalence of bacteria in fruits and vegetables. As a result, the study suggests monitoring and surveillance of fruit and vegetable sellers, national vendor hygiene policies and public awareness.

Extraction of Phenolic Compounds from Tucuma (Astrocaryum vulgare Mat.) Postprocessing Cake Mediated by Rhamnolipids: Evaluation of Its Antifungal Potential against Common Fruit Pathogens

Extraction of Phenolic Compounds from Tucuma (<i>Astrocaryum vulgare</i> Mat.) Postprocessing Cake Mediated by Rhamnolipids: Evaluation of Its Antifungal Potential against Common Fruit Pathogens

Biocontrol efficacy of Botrytis cinerea on postharvest tomato fruit by the endophytic bacterium Bacillus velezensis BE1

Endophytic bacteria offer numerous advantages in the control of postharvest diseases. Bacillus velezensis has been identified as a promising biocontrol agent against postharvest pathogens in various fruits and vegetables. In this study, B. velezensis BE1 was identified by 16S rRNA sequencing and caused 76.9 % inhibition against Botrytis cinerea in dual culture assay. The cell-free culture filtrate (CFCF) of B. velezensis strongly inhibited the growth, spore germination, and germ tube elongation of B. cinerea. Moreover, 11 bioactive compounds were identified in the CFCF using gas chromatography-mass spectrometry, and phthalic acid, dioctyl ester was the predominant component by 72.92 %. Scanning and transmission electron microscopy observations revealed that the CFCF causes irreversible morphological and ultrastructural damage to the fungal hyphae and conidia, including deformed, wrinkled, lysis, collapse, and degradation of cytoplasmic organelles. The incidence and severity of gray mold were significantly suppressed after applying CFCF at 50 % in tomato fruit. The application of CFCF caused an increase in the total phenolics and flavonoids content in tomato fruit compared with the control throughout the experiment. Furthermore, the activities of enzymes associated with defense such as phenylalanine ammonia-lyase, polyphenoloxidase, and peroxidase were significantly higher in the treated tomatoes relative to the untreated group. These results indicate that B. velezensis BE1 has significant potential as an effective biocontrol agent against the pathogen B. cinerea, offering a valuable approach for controlling gray mold in postharvest fruit.

Effect of temperature on in vitro germination and growth of Colletotrichum fioriniae, a new emerging pathogen of olive fruits.

Olive anthracnose induced by different Colletotrichum species causes dramatic losses of fruit yield and oil quality. The increasing incidence of Colletotrichum fioriniae (Colletotrichum acutatum species complex) as causal agent of olive anthracnose in Italy, is endorsing new studies on its biology, ecology, and environmental factors such as temperature. Five isolates from different sampling sites in Lazio region (Central Italy) were studied under controlled laboratory conditions aiming to better understand the differences of thermal development among the isolates and to lay the foundations of a future mathematical model able to describe the key aspects of the pathogen's life cycle. The mycelial growth rate and the conidial germination rate were assessed at seven different constant temperatures (5, 10, 15, 20, 25, 30, and 35°C) and fixed relative humidity (100% RH). The obtained dataset was analysed to estimate the parameters of mathematical functions that connect the mycelial growth rate and the spore germination with the environmental temperature. The parameters set provided as the result of this study constitute a key step forward in the biological knowledge of the species and the basis for future formulations of mathematical models that might be the core of decision support systems in an integrated pest management framework.

Rapid detection of Impatiens necrotic spot virus from thrips vectors using reverse transcription-recombinase polymerase amplification

The plant virus, Impatiens necrotic spot virus (INSV), is an economically important pathogen of vegetables, fruits, and ornamental crops. INSV is vectored by the western flower thrips, Frankliniella occidentalis, a small insect pest that is globally distributed. In recent years, INSV outbreaks have reached epidemic levels in the Salinas Valley of California—an agriculturally rich region where most of the lettuce (Lactuca sativa) is produced in the United States. Due to the obligate nature in which virus transmission occurs, new tools that could rapidly detect INSV from thrips vectors would enhance our ability to predict where virus outbreaks may occur. Here, we report on the development of a reverse transcription-recombinase polymerase amplification (RT-RPA) assay that can detect INSV from individual thrips. The assay uses crude extraction methods, is performed at a single temperature of 42 °C, can be completed in 25 min, and provides sensitivity levels that are comparable to other available detection methods. When the assay was used on field populations of thrips, INSV was successfully identified and quantified from individual larvae and adults. The work provides a new cost-effective surveillance tool that can rapidly detect INSV from its insect vector and from plants.

Volatile Organic Compounds Produced by Bacillus sp. Strain R2 Inhibit Aspergillus flavus Growth In Vitro and in Unhulled Rice.

Volatile organic compounds (VOCs) produced by Bacillus species exhibit biocontrol activity against fungal pathogens of fruits and vegetables. However, research on the effect of VOCs on Aspergillus flavus in stored grains is limited. This study aimed to investigate the effects of VOCs extracted from the strain R2, which was isolated from unhulled rice and identified as Bacillus paramycoides on A. flavus in vitro and unhulled rice. R2 VOCs effectively inhibited conidial germination and the hyphal growth of A. flavus in vitro. Moreover, R2 VOCs reduced the fungal population, aflatoxin B1 (AFB1) levels, and free fatty acid (FFA) value by 90.8%, 67%, and 38.7%, respectively, in unhulled rice. Eighteen R2 VOCs were identified using headspace solid-phase micro-extraction gas chromatography-mass spectrometry, and the individual activity of the VOCs against A. flavus was tested in vitro. Benzaldehyde (Ben) and 3,7-dimethyl-1-octanol (Dmo) showed strong inhibitory activities against A. flavus on PDA plates, with inhibition rates of 100% and 91.2%, respectively, at a concentration of 20 μL/dish. Ben at the concentration of 0.09 mg/mL, Dmo at the concentration of 0.07 mg/mL, or a mixture of both at halved concentrations could reduce the fungal population, AFB1 levels, and FFA content in unhulled rice. Our findings suggest that R2 VOCs are good alternatives to traditional chemical fumigants for suppressing A. flavus in stored grains. However, further research is necessary to establish the optimal fumigation concentration of these two components in unhulled rice. The impact of their residues on grain quality should be explored through sensory evaluation and nutritional analysis, and their safety to the environment and human body should be evaluated through safety assessment.

Green Silver Nanoparticles and Their Activity as Antifungal Agents Against Penicillium digitatum Causing Green Mould of Citrus

AbstractThe green synthesis of silver nanoparticles (SNPs) using aqueous extracts of five plants viz., Ocimum americanum, Azadirachta indica, Allium sativum, Eucalyptus hybrida, Zingiber officinale were evaluated for their antifungal property against Penicillium digitatum, a green mould pathogen of citrus fruits. The five plant species were selected from a screening of aqueous extracts from 16 plant species. The efficacy of aqueous extracts, at 50 % concentration, ranged from 14.44–68.52 % inhibition in mycelia growth of P. digitatum. Among SNPs, O. americanum was found most efficacious, at 1000 ppm concentration, with 80.7 % mycelial inhibition followed by A. indica with 77.4 % mycelial growth inhibition of fungal pathogen. Further, physical and structural characterization of SNPs of O. americanum and A. indica was carried out. The characterization analysis executed via transmission and scanning electron microscope revealed size distribution and nanoparticles were spherical to irregularly shaped ranging from 22.8–250 nm in O. americanum and 26.5–500 nm in A. indica. In UV‐visible spectrophotometer, the absolute absorption peak was recorded at 448 nm and 442 nm for O. americanum and A. indica, respectively. FTIR analysis of SNPs of A. indica and O. americanum confirmed the presence of carboxyl, amide group and secondary amines which were the main constituent of proteins. The novelty of the present study is in the green synthesis of silver nanoparticles with potential in the management of postharvest pathogens of fruit crops. The synthesis of SNPs used in this study is simple, environmentally benign and cost‐effective using aqueous extract of native plants.

Bacillus Species’ Gaseous Metabolites Prolong the Shelf Life of Apples and Grapes

In view of the threat to the environment posed by the widely used plant and fruit preservation chemicals in current agricultural practices, and the indispensability of fruit and plant preservatives in ensuring the yield of agricultural products, it is urgent to explore environmentally friendly and efficient preservative alternatives. Against this backdrop, bacterial preservatives stand out as a potential green solution. In this study, 12 strains of Bacillus derived from different ecological niches were carefully screened to evaluate the inhibitory efficacy of their gaseous metabolites against Anthracnose colloidospora, a common fruit pathogen, by bisectional method. The results showed that the bacterial suspension of strain B4 was the most prominent in inhibiting the growth of Colloidosporum anthracnose, and it significantly narrowed the growth circle of pathogenic bacteria, so it was selected as the object for further optimization and application. Subsequently, we designed a practical application test for fruit preservation, in which the bacteria of strain B4 were directly placed in the same environment as the fruit to be preserved, and the observation was continued for five days. By comparing and analyzing the fruit state before and after treatment, we were pleased to find that strain B4 showed a positive effect on inhibiting fruit rot, indicating its great potential as a natural fruit preservative. This discovery not only provides a scientific basis for reducing the use of chemical preservatives, but also contributes to the sustainable development of agriculture and the protection of the ecological environment.

Coffee Fruit Rot: The Previously Unrecognized Role of Fusarium and Its Interactions with the Coffee Berry Borer (Hypothenemus hampei).

Coffee fruit rot (CFR) is a well-known disease worldwide, mainly caused by Colletotrichum spp., the most important species being C. kahawae subsp. kahawae. In Puerto Rico, Colletotrichum spp. were identified as pathogens of coffee fruits. The coffee berry borer (CBB) was shown to be a dispersal agent of these fungi, and interaction of Fusarium with Colletotrichum affecting coffee fruits was suggested. In this study, we demonstrated that Fusarium spp. also cause CFR in Puerto Rico. Fusarium spp. are part of the CBB mycobiota, and this insect is responsible for spreading the pathogens in coffee fields. We identified nine Fusarium spp. (F. nirenbergiae, F. bostrycoides, F. crassum, F. hengyangense, F.solani-melongenae, F.pseudocircinatum, F. meridionale, F. concolor, and F. lateritium) belonging to six Fusarium species complexes isolated from CBBs and from rotten coffee fruits. Pathogenicity tests showed that F.bostrycoides, F. lateritium, F. nirenbergiae, F. solani-melongenae, and F.pseudocircinatum were pathogens causing CFR on green coffee fruits. F.bostrycoides was the predominant species isolated from the CBB mycobiota and coffee fruits with symptoms of CFR, suggesting a close relationship between F. bostrycoides and the CBB. To our knowledge, this is the first report of F. bostrycoides, F. solani-melongenae, F. pseudocircinatum, and F.nirenbergiae causing CFR worldwide and the first report of F. lateritium causing CFR in Puerto Rico. Understanding the CFR disease complex and how the CBB contributes to dispersing different Fusarium spp. on coffee farms is important to implement disease management practices in Puerto Rico and in other coffee-producing countries.

Molecular based associations of bacterial pathogens with citrus: Implications for toxicology and disease management

The losses associated with the bacterial diseases of citrus leaves and fruits have been reported throughout all of Pakistan's citrus-growing regions and a lot of work was done on the morphological characterization of pre and post-harvest pathogens. However, this issue has not been addressed in Khanpur, which is a major citrus-producing city in Pakistan and can help in the economic growth of the country. The citrus fruits in the area under study exhibit disease symptoms during pre and post-harvesting, but the actual disease agents have not yet been identified. The retrospective object of the current research is to study the pathogenic variation along with the investigation of bacterial disease incidence i.e. citrus canker, bacterial spots, & HLB in citrus fruit. During a survey conducted in November 2019 and 2020, bacterial isolates from symptomatic citrus fruits were obtained from randomly chosen orchards in Khanpur.The disease assessment was estimated by determining prevalence, severity, and disease incidence (DI). The bacterial isolates were identified morphologically and further through 16S rRNA gene amplification. The identified species were Pseudomonas syringae, Pseudomonas viridiflava, Serratia marcescens, Xanthomonas axonopodis pv citri, and Candidatus liberibacter. In the pathogenicity test, the Xanthomonas strains responsible for the canker disease caused the most severe disease symptoms. To our knowledge, this is the first report on pathogens of citrus fruits in Khanpur. Given the significant contribution of citrus fruits to household income and foreign exchange in Pakistan, this study illustrates the molecular-based association of bacterial pathogens of citrus and the toxic behavior of pathogens which helps to provide recommendations on genetic testing and to develop management programs for disease management. The molecular identification of citrus pathogens will empower the citrus growers, researchers, and regulatory agencies to make informed decisions that will protect citrus crops, reduce economic losses, and promote sustainable agriculture. It is also likely to play an increasingly vital role in the future of citrus cultivation..

The reproductive biology of Fagaceae acorns in the current and future climate

Fagaceae is an iconic plant family with 1000+ species distributed predominantly in the northern hemisphere. The reproductive ecology of the Fagaceae species is highly complex, and in the past three centuries, understanding the fruit masting strategies, dispersal, pathogen pressure, and storage has enjoyed steady research interest. However, the interrelationship between these factors is surprisingly less explored, undermining how acorns survive the post-dispersal period. The ability of fruits (acorns, nuts) to survive post-dispersal conditions starts during the maturation drying stage. Successful fruit production depends on the maternal environment, with a ‘good fruit production year’ occurring every few years. This phenomenon might out-maneuver predators by providing sufficient numbers to be consumed but still leaving enough for germination. Because most Fagaceae species have desiccation-sensitive (DS) fruits, survival after dispersal is challenging due to frequent dry spells. Fruit size is not phylogenetically conserved within the genus, with the average fruit mass of Castanea being 50 times higher than the desiccation-tolerant genus Fagus. Larger fruit size may be an adaptation in dry areas because prolonged drying continuously desiccates large-sized cotyledons before desiccating embryos. For species adapted to dry environments, synchronizing dispersal with the growing season could be beneficial, but exceptions exist. Many consumers, such as rodents, aid in the dispersal of fruits, but some fruits are killed in the process, especially those predated late. Upon settling on a site, the pericarp protects the embryo to a certain extent; if drying occurs, the cotyledons become the first site of water loss. However, under extreme drying, survival depends on sporadic rainfall, i.e., wet-dry cycle, which aids in maintaining the critical moisture content required by the fruits for survival. Nonetheless, these adaptive mechanisms are challenged by climate change, which affects the maturation, persistence, and seedling establishment of numerous Fagaceae species.

Efficacy and Synergistic Potential of Cinnamon (Cinnamomum zeylanicum) and Clove (Syzygium aromaticum L. Merr. & Perry) Essential Oils to Control Food-Borne Pathogens in Fresh-Cut Fruits.

The presence of microbial pathogens in ready-to-eat produce represents a serious health problem. The antibacterial activity of cinnamon (Cinnamomum zeylanicum) and clove (Syzygium aromaticum L. Merr. & Perry) essential oils (EOs) was determined toward food-borne pathogens by agar disk diffusion and minimum inhibitory concentration (MIC) assays. The growth kinetics of all strains, both in a buffer suspension assay and "on food" in artificially contaminated samples, were also investigated. The two EOs demonstrated a good antibacterial effect both alone and in combination (EO/EO). The use of EO/EO led to a synergistic antibacterial effect, also confirmed by the growth kinetics studies, where the EOs were active after 10 h of incubation (p < 0.0001) at significantly lower concentrations than those when alone. In the "on food" studies performed on artificially contaminated fruit samples stored at 4 °C for 8 days, the greatest killing activity was observed at the end of the trial (8 days) with a reduction of up to 7 log CFU/g compared to the control. These results confirm the good antibacterial activity of the EOs, which were more effective when used in combination. Data from the "on food" studies suggest cinnamon and clove essential oils, traditionally used in the food industry, as a possible natural alternative to chemical additives.

Bioinformatics-based identification of GH12 endoxyloglucanases in citrus-pathogenic Penicillium spp

Millions of tons of citrus peel waste are produced every year as a byproduct of the juice industry. Citrus peel is rich in pectin and xyloglucan, but while the pectin is extracted for use in the food industry, the xyloglucan is currently not valorized. To target hydrolytic degradation of citrus peel xyloglucan into oligosaccharides, we have used bioinformatics to identify three glycoside hydrolase 12 (GH12) endoxyloglucanases (EC 3.2.1.151) from the citrus fruit pathogens Penicillium italicum GL-Gan1 and Penicillium digitatum Pd1 and characterized them on xyloglucan obtained by alkaline extraction from citrus peel. The enzymes displayed pH-temperature optima of pH 4.6–5.3 and 35–37°C. PdGH12 from P. digitatum and PiGH12A from P. italicum share 84% sequence identity and displayed similar kinetics, although kcat was highest for PdGH12. In contrast, PiGH12B from P. italicum, which has the otherwise conserved Trp in subsite −4 replaced with a Tyr, displayed a 3 times higher KM and a 4 times lower kcat/KM than PiGH12A, but was the most thermostable enzyme of the three Penicillium-derived endoxyloglucanases. The benchmark enzyme AnGH12 from Aspergillus nidulans was more thermally stable and had a higher pH-temperature optimum than the enzymes from Penicillum spp. The difference in structure of the xyloglucan oligosaccharides extracted from citrus peel xyloglucan and tamarind xyloglucan by the new endoxyloglucanases was determined by LC-MS. The inclusion of citrus peel xyloglucan demonstrated that the endoxyloglucanases liberated fucosylated xyloglucan oligomers, implying that these enzymes have the potential to upgrade citrus peel residues to produce oligomers useful as intermediates or bioactive compounds.

Mitigating postharvest quantitative and qualitative losses in mango fruits through the application of biocontrol agents: An in-vivo and in-vitro assessment

Mango is a commercial fruit crop of India that suffers huge postharvest losses every year. The application of biocontrol agents (BCAs) bears a vast potential for managing the same, which is yet to be exploited to its fullest extent. Hence, studies were conducted for BCAs application of Debaryomyces hansenii, Bacillus subtilis and Pseudomonas fluorescens strains on mango fruit under in-vitro, in-vivo conditions to know the efficacy of these BCAs on the postharvest pathogen, shelf life and quality retention of mango fruit. The ‘poisoned food technique’ was attempted for in-vitro studies. For the in-vivo studies, fruit of the commercial cultivar ‘Amrapali’ were un-inoculated and pre-inoculated with major postharvest pathogens (anthracnose: Colletotrichum gloeosporioides and stem-end rot: Botryodiplodia theobromae) were treated with BCA, followed by ambient storage at (24 ± 4 °C, 75 ± 5 % RH). From the results, it has been observed that under in vitro studies, BCA Debaryomyces hansenii (Strain: KP006) and Bacillus subtilis (Strain: BJ0011) at the treatment level 108 CFU mL−1 while, the Pseudomonas fluorescens at 109 CFU mL−1 (Strain: BE0001) were significantly effective for pathogen inhibition. However, under the in vivo studies, the BCA Debaryomyces hansenii (Strain: KP006) at 108 CFU mL−1 treatment level was found to significantly reduce the pathogen's decay incidence while positively influencing the shelf life and biochemical (quality) attributes. This treatment increased the storage life of mango fruit by more than three days over control fruit. Therefore, BCA Debaryomyces hansenii (Strain: KP006) at 108 CFU mL−1 can be used to control the postharvest pathological loss of mango fruit without affecting its internal quality.

Origanum dictamnus Essential Oil in Vapour or Aqueous Solution Application for Pepper Fruit Preservation against Botrytis cinerea

The use of synthetic sanitizers for fresh commodities preservation is of concern, with eco-friendly alternatives, including essential oils (EOs), attracting research and industry interest. Dittany (Origanum dictamnus—DIT) oil was applied, either through vapour or dipping, on pepper fruit or in vitro against Botrytis cinerea, and compared to untreated (control) or chlorine (CHL)-treated fruits stored at 11 °C. Direct DIT vapours (up to 6 d) suppressed lesion growth, spore germination, and spore production compared to the untreated fruits. The antimicrobial properties of EOs were evidenced in pre-exposed-to-DIT oil vapours (residual effect), resulting in fruit lesion suppression. However, DIT-pretreated fruits had the same spore production and spore germination as the control and CHL applications. In in vitro tests, DIT vapours decreased colony growth and spore production when fungi were grown on Potato Dextrose Agar (PDA) or PDA pre-exposed to DIT following B. cinerea inoculation. This evidenced that the disease suppression after DIT vapour application primarily affected the interaction of the fruit–pathogen and/or residual responses on fruit tissue and/or PDA media. Fungal biomass in Potato Dextrose Broth (PDB) was evaluated after DIT and CHL applications (10–50–100–500–5000 μL L−1) and decreased with the DIT oil. Additionally, DIT or CHL sanitary dipping on pepper was ineffective against B. cinerea compared to vapour application. Therefore, DIT vapours revealed antimicrobial properties and could be an alternative postharvest sanitiser. DIT oil application should also be evaluated at semi-commercial scale for further optimizations, prior to commercialisation.

Volatile organic compounds as potential biomarkers of Cadophora luteo‐olivacea presence on kiwifruits

AbstractCadophora luteo‐olivacea is the causal agent of the skin‐pitting disease of kiwifruit, a syndrome that appears after 4–5 months of cold storage. However, it is assumed that the infection takes place in the field during fruit development. The present work takes into consideration the production of volatile organic compounds (VOCs) of Actinidia deliciosa ‘Hayward’ at different phenological phases as potential C. luteo‐olivacea infection biomarkers. In vitro assays were conducted to gain knowledge on the effect of kiwifruit VOCs on pathogen conidial germination and mycelial growth. VOCs produced by kiwifruit either inoculated or not with C. luteo‐olivacea were analysed at different phenological phases by SPME/GC–MS analysis. In particular, ethanol, o‐xylene, d‐limonene and acetic acid showed a significant increase in the presence of fungal inoculation. Ethanol and d‐limonene were also detected as volatile metabolites of the pathogen. The effect of each compound (ethanol, o‐xylene, d‐limonene and acetic acid) was tested on the fungal conidial germination at different concentrations, showing a growth stimulation at lower amounts. These results show how the production of some VOCs can contribute to the knowledge of fruit–pathogen interaction in the field with the aim of developing future tools for early disease detection and consequent effective control.

A circular economy approach: A new formulation based on a lemon peel medium activated with lactobacilli for sustainable control of post-harvest fungal rots in fresh citrus fruit

The aim of the study was to develop an effective bio-formulation for preventing post-harvest fungal rots in fresh citrus fruit through a multi-step process that includes isolation and characterization of lactic acid bacteria (LABs) from citrus fruit peel, selection of LAB strains with high antifungal activity, chemical characterization of cell free supernatants (CFS), formulation of a lemon peel powdery-based medium (LM) activated with LAB fermentates, chemical characterization of fermented LM, and evaluation of the efficacy of the new bio-formulation against blue mold. Thirteen LABs were recovered from the rind of citrus fruit and identified by the peptide mass fingerprinting method. The antifungal activity of LABs, isolated from citrus fruit, as well as cell free supernatants (CFSs), obtained from their liquid cultures, was tested against diverse plant pathogenic fungi and oomycetes, using the dual-culture overlay and the diffusion agar assays, respectively. Two isolates designated as N3B2 and M2B2, both identified as Lactiplantibacillus plantarum, were selected among the others for their relevant antifungal activity. Both isolates showed a broad spectrum antagonistic activity comprising also major postharvest pathogens of citrus fruit, such as Penicillium digitatum and P. italicum, the causal agents of green and blue moulds, respectively, Alternaria alternata, Colletotrichum gloeosporioides, C. karsti, Phytophthora citrophthora and Ph. nicotianae. The N3B2 and M2B2 isolates were used as starter cultures to ferment a lemon peel powder medium enriched with a nutrient water solution (LM). The two formulations obtained by fermenting LM with the N3B2 and M2B2 isolates showed a strong in vitro inhibitory activity on the same broad range of pathogens used in the preliminary screening of LABs. Moreover, the two LM-based formulations reduced the severity of blue mold infection and inhibited the sporulation of P. italicum on artificially inoculated lemon fruit. The chemical analysis of LM-based bioformulations revealed their antifungal activity was very probably due to the LAB-derived acidic components, including lactic, acetic, DL-3-phenyllactic, 3-4-dihydroxyhydrocinnamic, salicylic acid vanillic acids. These innovative LM-based bioformulations activated with LAB-derived antifungal compounds will be exploited as edible and biodegradable fruit coatings to prolong the shelf-life and prevent post-harvest rots of fresh citrus fruit.

Antimicrobial-resistant pathogens in fruits and vegetables from retail and home gardens

Abstract Fruits and vegetables have been identified as vehicles for the transmission of pathogenic antimicrobial-resistant (AMR) microorganisms. This is of food safety concern, thus requiring continuous surveillance. This study aimed to profile AMR bacteria present in selected fruits and vegetables retailed in markets and from home gardens (HGs) in Oyo, Ogun, and Ekiti states, Nigeria. Watermelon, cucumber, tomato, and garden egg samples were collected and analyzed using standard microbiological procedures. The susceptibility of the isolates to eight antibiotics was determined. Multidrug-resistant isolates were screened for the presence of AMR genes by polymerase chain reaction. Fifty three bacteria were isolated and identified, belonging to the genera Bacillus, Corynebacterium, Listeria, Aeromonas, Enterobacter, Erwinia, Salmonella, Serratia, Shigella, and Vibrio. Thirty six (67.93%) isolates demonstrated phenotypic resistance to five of the eight antibiotics tested, being the most prevalent pattern observed. BlaTEM and blaCTX-M were detected in Salmonella enterica from retailed tomato, blaTEM, blaSHV, blaCTX-M, and erm(B) were detected in Listeria monocytogenes from retailed watermelon, blaSHV and blaCTX-M were detected in Bacillus cereus from retailed tomato, while blaTEM, blaSHV, blaCTX-M, and erm(F) were detected in Staphylococcus aureus isolated in garden egg from HG. The presence of multidrug-resistant pathogens in fruits and vegetables could pose a huge food safety and public health risk.

Exploring the Biocontrol Capability of Non-Mycotoxigenic Strains of Penicillium expansum.

Penicillium expansum is one the major postharvest pathogens of pome fruit during postharvest handling and storage. This fungus also produces patulin, which is a highly toxic mycotoxin that can contaminate infected fruits and their derived products and whose levels are regulated in many countries. In this study, we investigated the biocontrol potential of non-mycotoxigenic strains of Penicillium expansum against a mycotoxigenic strain. We analyzed the competitive behavior of two knockout mutants that were unable to produce patulin. The first mutant (∆patK) involved the deletion of the patK gene, which is the initial gene in patulin biosynthesis. The second mutant (∆veA) involved the deletion of veA, which is a global regulator of primary and secondary metabolism. At the phenotypic level, the ∆patK mutant exhibited similar phenotypic characteristics to the wild-type strain. In contrast, the ∆veA mutant displayed altered growth characteristics compared with the wild type, including reduced conidiation and abnormal conidiophores. Neither mutant produced patulin under the tested conditions. Under various stress conditions, the ∆veA mutants exhibited reduced growth and conidiation when exposed to stressors, including cell membrane stress, oxidative stress, osmotic stress, and different pH values. However, no significant changes were observed in the ∆patK mutant. In competitive growth experiments, the presence of non-mycotoxigenic strains reduced the population of the wild-type strain during in vitro growth. Furthermore, the addition of either of the non-mycotoxigenic strains resulted in a significant decrease in patulin levels. Overall, our results suggest the potential use of non-mycotoxigenic mutants, particularly ∆patK mutants, as biocontrol agents to reduce patulin contamination in food and feed.

The low-affinity calcium channel protein AaFig1 is essential for the growth and development, infection structure differentiation, and pathogenicity of Alternaria alternata

Fig1, as a low-affinity Ca2+ channel, is needed for Ca2+ influx and normal mating found in a few species, but its detailed molecular regulating mechanism in fungi, especially in Alternaria alternata, the causal agent of pear black spot, is still unclear. In this study, Fig1 in A. alternata was cloned, identified and named as AaFig1, and its function was characterized by targeted gene knockout strategy. The results showed that the ΔAaFig1 was severely decreased in vegetative growth, biomass accumulation and sporulation. Fluo-4, AM staining indicated that the Ca2+ content in ΔAaFig1 was decreased. Deletion of AaFig1 reduced the response of A. alternata to exogenous Ca2+ but improved the tolerance to osmotic stresses. Interestingly, the appressorium formation of the ΔAaFig1 strain on hydrophobic and pear wax extract coated surface, and infection hyphae formation on the onion epidermis were significantly reduced, meanwhile, the ΔAaFig1 strain could not penetrate the cellophane. In addition, virulence on wound-inoculated pear fruit and intact pear leaves, melanin production and the activities of cell wall degrading enzymes in AaFig1 strain were reduced. In summary, these results indicate that AaFig1-mediated Ca2+ homeostasis positively regulates the growth and development, infection structure formation and pathogenicity of A. alternata. The findings will provide a basis for developing new strategies to control this pathogen in pear fruit.