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Microbiol. Res., Volume 15, Issue 3 (September 2024) – 13 articles

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17 pages, 683 KiB  
Article
New Insights into Molecular Characterization, Antimicrobial Resistance and Virulence Factors of Methicillin-Sensitive Coagulase-Positive Staphylococcus spp. from Dogs with Pyoderma and Otitis Externa
by Faten Ben Chehida, Wafa Tombari, Haythem Gharsa, Youssef Rabia, Sana Ferhi, Maha Jrad and Lilia Messadi
Microbiol. Res. 2024, 15(3), 1208-1224; https://doi.org/10.3390/microbiolres15030081 (registering DOI) - 12 Jul 2024
Abstract
The first Tunisian national molecular survey of coagulase-positive staphylococci (CoPS) isolated from dogs with pyoderma and otitis externa was conducted to evaluate the prevalence of CoPS and identify its phenotypic and genotypic diversities. A total of 99 out of the 195 samples collected [...] Read more.
The first Tunisian national molecular survey of coagulase-positive staphylococci (CoPS) isolated from dogs with pyoderma and otitis externa was conducted to evaluate the prevalence of CoPS and identify its phenotypic and genotypic diversities. A total of 99 out of the 195 samples collected from 39 sick dogs were identified across multiple sites as methicillin-susceptible CoPS belonging to the species S. pseudintermedius (64.4%), S. aureus (20.2%), S. coagulans (10.1%), and S. hyicus (5%). Fifteen sampled dogs carried more than one Staphylococcus species. Their antibiotic resistance and virulence factors were determined using conventional and molecular methods. Of the S. pseudintermedius isolates found, 17.4% were multidrug-resistant, whereas high rates of virulence genes were observed among the S. aureus isolates. On polystyrene surfaces, 75% of S. aureus isolates were biofilm producers, of which 15% were classified as strong producers. The capsular polysaccharide cap8 genotype was predominant among them. A MultiLocus Sequence Typing (MLST) analysis clustered the S.aureus isolates into five distinct sequence types (STs), with four assigned for the first time. Our findings highlight the spread of CoPS among diseased dogs and, especially, the emergence of S. hyicus, S. coagulans, multidrug-resistant S. pseudintermedius and S. aureus isolates with high genetic variability. The precise characterization of these strains, as well as their continuous monitoring, is necessary for the implementation of preventive strategies given the significant public health risk. Full article
(This article belongs to the Special Issue Veterinary Microbiology and Diagnostics)
19 pages, 1659 KiB  
Article
Predictive Modeling of UV-C Inactivation of Microorganisms in Glass, Titanium, and Polyether Ether Ketone
by Amira Chroudi, Talita Nicolau, Narayan Sahoo, Óscar Carvalho, Andrea Zille, Samir Hamza and Jorge Padrão
Microbiol. Res. 2024, 15(3), 1189-1207; https://doi.org/10.3390/microbiolres15030080 - 5 Jul 2024
Viewed by 423
Abstract
Biomaterials consist of both natural and synthetic components, such as polymers, tissues, living cells, metals, and ceramics. Their purpose is focused on repairing or replacing malfunctioning living tissues and organs. Therefore, it is imperative to ensure the safety and sterility of biomaterials before [...] Read more.
Biomaterials consist of both natural and synthetic components, such as polymers, tissues, living cells, metals, and ceramics. Their purpose is focused on repairing or replacing malfunctioning living tissues and organs. Therefore, it is imperative to ensure the safety and sterility of biomaterials before any contact with living tissue. Ultraviolet (UV)-C irradiation for biomaterial disinfection has been considered due to the high recurrence rate of bacterial infections and to prevent resistance. Physical composition and surface properties and UV-C sensitivity of microorganisms can alter its efficacy. The main objective of this study was to evaluate the efficacy of UV-C in terms of microbial lethality and additional underlying factors contributing to its performance, namely the surface properties. For this purpose, twelve different strains were first screened, from which four microorganism species known to have the ability to cause nosocomial infections were further tested, namely Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and Candida glabrata. These microorganisms were inoculated onto slides and disks of various bio contact surfaces, including glass (GLS), titanium (Ti), and poly ether etherketone (PEEK), and exposed to UV-C. The results demonstrate that bacterial pathogens on biomaterial surfaces respond differently to UV-C light exposure, and the bactericidal effect decreased in this order: glass, PEEK, and Ti (0.5 to 2.0 log reduction differences). P. aeruginosa ATCC 27853 on glass surfaces was reduced to an undetectable level after being exposed to 6.31 J.cm−2 of UV-C, displaying the highest reduction rate observed among all the tested microorganisms, 2.90 J−1.cm−3, compared to Ti and PEEK. Similarly, a higher reduction in C. glabrata ATCC 2001 was observed on glass; the modeled inhibition displayed a rate of 1.30 J−1.cm3, the highest observed rate among yeast, compared to Ti and PEEK, displaying rates of 0.10 J−1.cm−3 and 0.04 J−1.cm−3, respectively. The inactivation rates were higher for less hydrophobic materials with smoother surfaces as compared to biomaterials with rougher surfaces. Full article
11 pages, 5923 KiB  
Article
Limosilactobacillus reuteri and Its Probiotic Potential against Cariogenic Bacteria
by Brianda Karina Félix-Sicairos, Rita Elizabeth Martinez-Martinez, Saray Aranda-Romo, Roberto Gonzalez-Amaro, Mariana Salgado-Bustamante, Norma Verónica Zavala-Alonso and Ángel Gabriel Alpuche-Solís
Microbiol. Res. 2024, 15(3), 1178-1188; https://doi.org/10.3390/microbiolres15030079 - 3 Jul 2024
Viewed by 489
Abstract
Bacteriotherapy is a promising option in addressing dental caries, a persistent global public health challenge with multifactorial origin, including dysbiosis. Despite the exploration of various probiotics, outcomes remain inconclusive. Objective: This study aimed to assess the inhibitory potential of L. reuteri and other [...] Read more.
Bacteriotherapy is a promising option in addressing dental caries, a persistent global public health challenge with multifactorial origin, including dysbiosis. Despite the exploration of various probiotics, outcomes remain inconclusive. Objective: This study aimed to assess the inhibitory potential of L. reuteri and other potential probiotics like S. salivarius and S. oralis on the growth, adhesion, colonization, and viability of major cariogenic pathogens, comparing their probiotic efficacy. Methods: An in vitro experimental study was conducted, encompassing direct competition assays in solid and liquid co-culture tests and the characterization of adhesion to dental enamel and cell viability by life or death assay. Results: L. reuteri exhibited the significant inhibition of S. sobrinus and S. mutans growth in both solid and liquid cultures, with statistically notable differences. Scanning electron microscopy and confocal microscopy demonstrated reduced cariogenic biofilm formation when combined with L. reuteri, corroborated by diminished bacterial viability and decreased dental enamel coverage. These findings underscore L. reuteri’s potential as an effective agent in caries prevention. Conclusion: The study suggests L. reuteri could serve as an effective probiotic in bacteriotherapy against dental caries. It displayed substantial inhibitory activity in vitro against cariogenic bacteria, impeding biofilm formation and adhesion, thereby impacting cell viability. Full article
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15 pages, 932 KiB  
Article
Comparative High-Throughput Sequencing Analysis of the Bacterial Community Associated with the Rhizosphere of Date Palm (Phoenix dactyllifera L.) Irrigated with Treated Wastewater and Groundwater
by Fayza Kouadri
Microbiol. Res. 2024, 15(3), 1163-1177; https://doi.org/10.3390/microbiolres15030078 - 2 Jul 2024
Viewed by 259
Abstract
Water scarcity is a major agricultural issue in most arid and semi-arid regions of the world. Alternative water supplies, such as the reuse of wastewater for agricultural irrigation, have been introduced. However, little is known about their impact on the soil and rhizosphere [...] Read more.
Water scarcity is a major agricultural issue in most arid and semi-arid regions of the world. Alternative water supplies, such as the reuse of wastewater for agricultural irrigation, have been introduced. However, little is known about their impact on the soil and rhizosphere microbiomes that receive irrigation. Therefore, this work evaluates the impact of treated wastewater (TWW) irrigation on the soil and rhizosphere bacterial communities of date palms in Al Madinah, Saudi Arabia. In this study, metagenomic DNA from the rhizosphere of the date palm was sequenced using Illumina high-throughput sequencing. According to the observed OTUs, Chao1 richness estimations, and Shannon diversity values, soils from groundwater-irrigated date palms showed higher microbial diversity than did soils from TWW-irrigated date palms. A total of 569 OTUs were generated; most of them (97.3%) were assigned into 15 different phyla, whereas 2.7% were marked as unclassified. DNA sequence analysis of the WWT-irrigated rhizosphere showed that the most abundant phyla were Firmicutes (43.6%), Bacteroidetes (17.3%), Proteobacteria (15.2%), and Actinobacteria (14.6%), representing more than 90.7% of the total community, while the soil of the rhizosphere irrigated with GW was dominated by Actinobacteria (44.1%), Proteobacteria (23.4%), Firmicutes (15.5%), and Gemmatimonadetes (4.9%). The most frequently observed species in the two soils were also different. The dominant species in TWW-irrigated soil was Planococcus plakortidis, which is prevalent in saline and moderately saline habitats and can play an important ecological role. The GW-irrigated rhizosphere exhibited higher levels of biocontrol bacteria, particularly Nocardioides mesophilus. These results provide a comprehensive understanding and insights into the population dynamics and microbiome of date palm rhizosphere. The findings show that the irrigation water quality has a significant impact on the microbiome composition. Identifying the microbial diversity is the first step toward determining the best way to use TWW in irrigation. Full article
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19 pages, 2421 KiB  
Article
Plant Growth-Promoting and Tequila Vinasse-Resistant Bacterial Strains
by Alberto J. Valencia-Botín, Ismael F. Chávez-Díaz, Florentina Zurita-Martínez, Allan Tejeda-Ortega and Lily X. Zelaya-Molina
Microbiol. Res. 2024, 15(3), 1144-1162; https://doi.org/10.3390/microbiolres15030077 - 2 Jul 2024
Viewed by 433
Abstract
Tequila vinasse, a byproduct of the tequila industry, is frequently discharged into water bodies or agricultural fields, posing significant ecological and human health risks. Bacterial communities that inhabit these agricultural fields have developed mechanisms to utilize tequila vinasse as a potential nutrient source [...] Read more.
Tequila vinasse, a byproduct of the tequila industry, is frequently discharged into water bodies or agricultural fields, posing significant ecological and human health risks. Bacterial communities that inhabit these agricultural fields have developed mechanisms to utilize tequila vinasse as a potential nutrient source and to promote plant growth. In this study, strains from the phyla Actinomycetota and Pseudomonadota were isolated from agricultural fields irrigated with tequila vinasse for 2, 10, and 14 years in Jalisco, Mexico. The results showed that strains of Terrabacter, Azotobacter, Agromyces, Prescottella, and Leifsonia tolerate high concentrations of tequila vinasse and promote maize seedling growth in the presence of tequila vinasse. Additionally, some of the strains solubilize potassium and produce siderophores, cellulase, protease, lipase, and esterase. The strains Terrabacter sp. WCNS1C, Azotobacter sp. WCNS1D, and Azotobacter sp. WCNS2A have potential applications in the bioremediation of tequila vinasse in agricultural fields discharged with tequila vinasse. Full article
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10 pages, 500 KiB  
Brief Report
Factors Influencing Central Venous Catheter-Associated Bloodstream Infections in COVID-19 Patients
by Adriana Lemos de Sousa Neto, Thalita Campos, Clesnan Mendes-Rodrigues, Reginaldo dos Santos Pedroso and Denise Von Dolinger de Brito Röder
Microbiol. Res. 2024, 15(3), 1134-1143; https://doi.org/10.3390/microbiolres15030076 - 2 Jul 2024
Viewed by 471
Abstract
During the pandemic of COVID-19, the rates of bloodstream infection associated with venous catheter in patients infected with the disease admitted to an intensive care unit rose significantly. In this study, we evaluated the occurrence of bloodstream infections in patients with SARS-CoV-2 and [...] Read more.
During the pandemic of COVID-19, the rates of bloodstream infection associated with venous catheter in patients infected with the disease admitted to an intensive care unit rose significantly. In this study, we evaluated the occurrence of bloodstream infections in patients with SARS-CoV-2 and the variables that made the patients more susceptible to the catheter-associated bloodstream infection (CABSI). Blood culture results from patients interned between March 2020 and December 2021 (n= 109) were collected electronically from the hospital information system and then analyzed. The following variables presented statistical relevance after an adjusted model as follows: obesity (p = 0.003) and time of use of catheter before infection (p = 0.019). In conclusion, patients with shorter catheter use time and obesity had higher incidence of CABSI. Full article
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15 pages, 1003 KiB  
Article
Characterization of Extended-Spectrum β-Lactamase Producing- and Carbapenem–Resistant Escherichia coli Isolated from Diarrheic Dogs in Tunisia: First Report of blaIMP Gene in Companion Animals
by Asma Ben Haj Yahia, Ghassan Tayh, Sarrah Landolsi, Ala Maazaoui, Faten Ben Chehida, Aymen Mamlouk, Monia Dâaloul-Jedidi and Lilia Messadi
Microbiol. Res. 2024, 15(3), 1119-1133; https://doi.org/10.3390/microbiolres15030075 - 30 Jun 2024
Viewed by 407
Abstract
Escherichia coli is an important opportunistic pathogen, causing several infections in dogs. The antimicrobial resistance of E. coli occurring in companion animals becomes an emerging problem. This study aimed to estimate the prevalence of ESBL-producing E. coli in diarrheic dogs, investigate the occurrence [...] Read more.
Escherichia coli is an important opportunistic pathogen, causing several infections in dogs. The antimicrobial resistance of E. coli occurring in companion animals becomes an emerging problem. This study aimed to estimate the prevalence of ESBL-producing E. coli in diarrheic dogs, investigate the occurrence and molecular characterization of carbapenem-resistant isolates, and determine their virulence genes. Fecal samples were collected from 150 diarrheic dogs in Tunisia. E. coli isolates were screened for antimicrobial resistance against 21 antibiotics by the disk diffusion method. The characterization of β-lactamase genes, associated resistance genes, and virulence genes was studied using PCR. Among 95 E. coli strains, 25 were ESBL-producing, and most of them were multidrug-resistant. The most prevalent β-lactamase genes were blaCTX-M1 (n = 14), blaTEM (n = 3), and blaCMY (n = 2). The blaIMP carbapenemase gene was found in two carbapenem-resistant isolates, which showed that carbapenemase-producing E. coli spread to companion animals in Tunisia. Different virulence genes associated with extraintestinal pathogenic E. coli were detected. This is the first report of the characterization of carbapenem resistance and virulence genes in dogs in North Africa. Our study showed that diarrheic dogs in Tunisia can be a potential reservoir of ESBL- or carbapenemase-producing E. coli with a possible risk of transmission to humans. Full article
(This article belongs to the Special Issue Zoonotic Bacteria: Infection, Pathogenesis and Drugs)
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9 pages, 2613 KiB  
Communication
Clonality Analysis of Streptococcus pneumoniae in Clinical Specimens
by Delly Chipta Lestari, Pranee Somboonthum, Daisuke Motooka, Eiji Ishii, Shigeaki Matsuda, Anis Karuniawati and Tetsuya Iida
Microbiol. Res. 2024, 15(3), 1110-1118; https://doi.org/10.3390/microbiolres15030074 - 29 Jun 2024
Viewed by 377
Abstract
Pneumococcal pneumonia is a significant cause of illness and death globally, particularly among young children and the elderly. The cpsB gene is involved in the biosynthesis of the capsule polysaccharide, and polymorphisms in the cpsB gene are the basis for sequetyping, a molecular [...] Read more.
Pneumococcal pneumonia is a significant cause of illness and death globally, particularly among young children and the elderly. The cpsB gene is involved in the biosynthesis of the capsule polysaccharide, and polymorphisms in the cpsB gene are the basis for sequetyping, a molecular biology-based approach to serotyping. In this study, we attempted the sequetyping of pneumococci directly from clinical sputum specimens collected from adult patients diagnosed with community-acquired pneumonia (CAP). We performed conventional PCR for the cpsB gene, followed by TA cloning and Sanger sequencing of the amplicon. The results showed the status of clonality of pneumococci in each specimen. We also performed real-time PCR targeting pneumococci for each specimen. It revealed a significant association between the Ct value of the real-time PCR and the clonality status of pneumococci among the specimens (p-value 0.0007 by Fisher’s exact test analysis). Specifically, when the Ct value was below 22, there was a high probability that pneumococcus existed as a single clone. Thus, this study demonstrates the possible correlation between pneumococcal clonality and bacterial load in clinical specimens, which might indicate the infection status. Full article
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11 pages, 1691 KiB  
Communication
Effect of Buffer Room Configuration on Isolation of Bacteriophage phi6 and Micrococcus Luteus Emissions
by Radosław Wróbel, Monika Andrych-Zalewska, Jędrzej Matla, Justyna Molska, Gustaw Sierzputowski, Agnieszka Szulak, Radosław Włostowski, Adriana Włóka and Małgorzata Rutkowska-Gorczyca
Microbiol. Res. 2024, 15(3), 1099-1109; https://doi.org/10.3390/microbiolres15030073 - 27 Jun 2024
Viewed by 258
Abstract
In light of the growing need to mitigate viral infections, such as SARS-CoV-2, which has caused over 770 million infections and 7 million deaths worldwide, the role of architectural design in reducing pathogen spread has become paramount. This study explores how different buffer [...] Read more.
In light of the growing need to mitigate viral infections, such as SARS-CoV-2, which has caused over 770 million infections and 7 million deaths worldwide, the role of architectural design in reducing pathogen spread has become paramount. This study explores how different buffer room configurations impact microbial isolation using the bacterium Micrococcus luteus and bacteriophage phi6 as test organisms. Through a comprehensive approach integrating architecture, technology, and scientific methodologies, we developed strategies to optimise safe buffer environments. Comparative analyses of various isolation systems highlighted significant variations in their effectiveness at reducing bioaerosol concentrations, directly influenced by the design and layout of buffer rooms. These findings are crucial for the effective design of medical and public spaces, particularly during pandemics, where preventing virus transmission is critical for public health. Full article
8 pages, 512 KiB  
Article
First Seroepidemiological Study of Toxoplasma gondii Infection in Dromedaries (Camelus dromedarius) in Southern Tunisia
by Afef Jeljli, Syrine Rekik, Boubaker Ben Smida, Walid Chandoul, Limam Sassi and Mohamed Gharbi
Microbiol. Res. 2024, 15(3), 1091-1098; https://doi.org/10.3390/microbiolres15030072 - 25 Jun 2024
Viewed by 652
Abstract
Toxoplasma gondii is one of the most common zoonotic parasites worldwide, with infections in humans as well as in all mammals, including dromedaries, that affect reproductive health, leading to malformations and abortions. In the current study, we estimated, for the first time in [...] Read more.
Toxoplasma gondii is one of the most common zoonotic parasites worldwide, with infections in humans as well as in all mammals, including dromedaries, that affect reproductive health, leading to malformations and abortions. In the current study, we estimated, for the first time in Tunisia, the seroprevalence of T. gondii antibodies and its associated risk factors in dromedaries (Camelus dromedarius) using the enzyme-linked immunosorbent assay (ELISA) technique. A total of 248 sera samples were collected from dromedaries living in Médenine and Tataouine governorates, South Tunisia. The overall seroprevalence of T. gondii was estimated to be 29.8 ± 2.9% (74/248) with a significantly higher seroprevalence rate in dromedaries from Médenine (52.5 ± 7.9%; 21/40) compared to Tataouine (25.5 ± 3%; 53/208); it was also higher in dromedaries kept in intensive farming systems (55 ± 11.1%; 11/20) compared to extensive systems (27.6 ± 3%; 63/228), in non-pregnant females (39 ± 4%; 57/146) compared to pregnant ones (16.7 ± 3.7%; 17/102), and in those in contact with cats (66.7 ± 8.6%; 20/30) compared to those with no contact with cats (24.8 ± 2.9%; 54/218). The present study should be followed by others to explore the role of dromedaries in the epidemiological cycle of T. gondii in Southern Tunisia. Full article
8 pages, 304 KiB  
Communication
Multidrug-Resistant Enterococcus faecium and Enterococcus faecalis Isolated from Dogs and Cats in Southern Brazil
by Letícia da Silva, Cristina Zaffari Grecellé, Ana Paula Guedes Frazzon, André Felipe Streck, Diéssy Kipper, André Salvador Kazantzi Fonseca, Nilo Ikuta and Vagner Ricardo Lunge
Microbiol. Res. 2024, 15(3), 1083-1090; https://doi.org/10.3390/microbiolres15030071 - 25 Jun 2024
Viewed by 536
Abstract
Enterococcus spp. are isolated from infections of domestic animals and can present resistance to different antimicrobials. This study aimed to identify the main species of Enterococcus and determine the phenotypic resistance to antimicrobials of isolates of this bacterial genus obtained from dogs and [...] Read more.
Enterococcus spp. are isolated from infections of domestic animals and can present resistance to different antimicrobials. This study aimed to identify the main species of Enterococcus and determine the phenotypic resistance to antimicrobials of isolates of this bacterial genus obtained from dogs and cats admitted to a veterinary hospital in southern Brazil. A total of 57 Enterococcus spp. were isolated from different clinical samples (urine, feces, ears and skin) in domestic animals admitted to the hospital over a period of three years (2016 to 2019). MALDI-TOF results demonstrated the occurrence of Enterococcus faecium (39; 68.4%), Enterococcus faecalis (17; 29.8%) and Enterococcus avium (1; 1.8%). In an overall analysis, resistance was observed to the antimicrobials rifampicin (46; 80.7%), tetracycline and streptomycin (42; 73.7%), ampicillin and imipenem (41; 71.9%), erythromycin (39; 68.4%), gentamicin (38; 66.7%), ciprofloxacin (36; 63.2%), norfloxacin (32; 56.1%), nitrofurantoin (10; 17.5%) and chloramphenicol (9; 15.7%). None of the Enterococcus spp. showed resistance to vancomycin and linezolid. Multidrug resistance (MDR) was detected in 45 (78.9%) isolates. In conclusion, E. faecium and E. faecalis with MDR are frequent in infections of hospitalized dogs and cats from southern Brazil. Full article
10 pages, 614 KiB  
Article
Comparative Mortality Analysis in Febrile and Afebrile Emergency Department Patients with Positive Blood Cultures: A Retrospective Study
by Nicola Bonadia, Davide Antonio Della Polla, Rita Murri, Tiziana D’Inzeo, Barbara Fiori, Annamaria Carnicelli, Andrea Piccioni, Mariella Fuorlo, Martina Petrucci, Angela Saviano, Antonio Gasbarrini, Francesco Franceschi and Marcello Covino
Microbiol. Res. 2024, 15(3), 1073-1082; https://doi.org/10.3390/microbiolres15030070 - 22 Jun 2024
Viewed by 337
Abstract
This retrospective analysis at a major Italian university hospital (January 2018–September 2022) assessed the prognostic significance of fever in patients with bloodstream infections (BSIs). Of the 1299 patients with positive blood cultures, a comparison between febrile and afebrile patients at emergency department admission [...] Read more.
This retrospective analysis at a major Italian university hospital (January 2018–September 2022) assessed the prognostic significance of fever in patients with bloodstream infections (BSIs). Of the 1299 patients with positive blood cultures, a comparison between febrile and afebrile patients at emergency department admission was conducted. This study particularly focused on the mortality rates associated with these two groups. Notably, afebrile patients exhibited a higher mortality rate. The odds ratio for mortality in afebrile patients was significantly higher compared to febrile patients. This suggests that the absence of fever might be an indicator of increased mortality risk, highlighting the complexity of diagnosing bloodstream infections based on fever presence. This study contributes to the understanding of fever as a diagnostic marker in emergency settings. Full article
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23 pages, 2444 KiB  
Article
Xylella fastidiosa Dispersion on Vegetal Hosts in Demarcated Zones in the North Region of Portugal
by Talita Loureiro, Luís Serra, Ângela Martins, Isabel Cortez and Patrícia Poeta
Microbiol. Res. 2024, 15(3), 1050-1072; https://doi.org/10.3390/microbiolres15030069 - 21 Jun 2024
Viewed by 671
Abstract
The detection of Xylella fastidiosa in Portugal occurred through prospection in 2019 in Vila Nova de Gaia, Porto, in plants of Lavandula dentata L. Currently, in Portugal, there are 18 Xylella fastidiosa Demarcated Zones (DZs). The main objective of this study is to [...] Read more.
The detection of Xylella fastidiosa in Portugal occurred through prospection in 2019 in Vila Nova de Gaia, Porto, in plants of Lavandula dentata L. Currently, in Portugal, there are 18 Xylella fastidiosa Demarcated Zones (DZs). The main objective of this study is to gain a comprehensive understanding of this problem within the defined Demarcated Zones in the North Region of Portugal from 2019 to June 2023. This work comprised two phases: the prospection phase (inspecting plants, sampling, and submission of samples to the laboratory) and the research phase (collecting and organizing data and statistical treatment). Our findings provide essential insights, suggesting that the Northern Region of Portugal is highly conducive to Xylella fastidiosa. Portugal has Xf-preferred hosts such as the olive tree, orange tree, Laurus nobilis, Rosa spp., Nerium oleander L., Pelargonium sp., Hedera helix L., and Lavandula dentata L. Portugal’s favorable environmental factors such as temperature and humidity can have an important role in influencing the interaction between bacteria and hosts. Nevertheless, it is crucial to consider variations in the behavior of insect vectors, as these variations can limit the prevalence of the disease. Activities like the transport of infected planting materials from the first Demarcated Zone of the Area Metropolitana do Porto could be associated with the introduction and spread of Xylella fastidiosa, potentially triggering new disease outbreaks in the country. Our findings confirm the alarming spread of Xylella fastidiosa across Portugal. Factors such as the presence of insect vectors, abundance of host plants, and climate suitability play pivotal roles in Xylella fastidiosa dispersal. We recommend that countries identified with relatively high risk, like Portugal, undertake thorough individual risk analyses. The implementation of preventive measures and, if necessary, the enhancement of surveillance systems for early detection of Xylella fastidiosa in plants and insect vectors are crucial steps. Full article
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