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PloS One 2021The Rimac river is the main source of water for Lima, Peru's capital megacity. The river is constantly affected by different types of contamination including mine...
The Rimac river is the main source of water for Lima, Peru's capital megacity. The river is constantly affected by different types of contamination including mine tailings in the Andes and urban sewage in the metropolitan area. In this work, we aim to produce the first characterization of aquatic bacterial communities in the Rimac river using a 16S rRNA metabarcoding approach which would be useful to identify bacterial diversity and potential understudied pathogens. We report a lower diversity in bacterial communities from the Lower Rimac (Metropolitan zone) in comparison to other sub-basins. Samples were generally grouped according to their geographical location. Bacterial classes Alphaproteobacteria, Bacteroidia, Campylobacteria, Fusobacteriia, and Gammaproteobacteria were the most frequent along the river. Arcobacter cryaerophilus (Campylobacteria) was the most frequent species in the Lower Rimac while Flavobacterium succinicans (Bacteroidia) and Hypnocyclicus (Fusobacteriia) were the most predominant in the Upper Rimac. Predicted metabolic functions in the microbiota include bacterial motility and quorum sensing. Additional metabolomic analyses showed the presence of some insecticides and herbicides in the Parac-Upper Rimac and Santa Eulalia-Parac sub-basins. The dominance in the Metropolitan area of Arcobacter cryaerophilus, an emergent pathogen associated with fecal contamination and antibiotic multiresistance, that is not usually reported in traditional microbiological quality assessments, highlights the necessity to apply next-generation sequencing tools to improve pathogen surveillance. We believe that our study will encourage the integration of omics sciences in Peru and its application on current environmental and public health issues.
Topics: Aquatic Organisms; Arcobacter; Computational Biology; DNA Barcoding, Taxonomic; Environmental Monitoring; Flavobacterium; Fusobacteria; Microbiota; Peru; RNA, Ribosomal, 16S; Rivers; Sewage; Water; Water Microbiology; Water Pollution
PubMed: 33886647
DOI: 10.1371/journal.pone.0250401 -
Revista Espanola de Quimioterapia :... Jun 2021
Topics: Arcobacter; Child; Humans; Spain; Species Specificity
PubMed: 33810639
DOI: 10.37201/req/134.2020 -
Antibiotics (Basel, Switzerland) Mar 2021spp. are emerging waterborne and foodborne zoonotic pathogens responsible for gastroenteritis in humans. In this work, we evaluated the occurrence and the antimicrobial...
spp. are emerging waterborne and foodborne zoonotic pathogens responsible for gastroenteritis in humans. In this work, we evaluated the occurrence and the antimicrobial resistance profile of isolates recovered from different aquatic sources. Besides, we searched for spp. in seaweeds and the corresponding seawater samples. Bacteriological and molecular methods applied to 100 samples led to the isolation of 28 isolates from 27 samples. The highest prevalence was detected in rivers followed by artificial ponds, streams, well waters, and spring waters. Seaweeds contained a higher percentage of than the corresponding seawater samples. The isolates were identified as (96.4%) and (3.6%). All the isolates showed a multi-drug resistance profile, being resistant to at least three different classes of antibiotics. Molecular analysis of genetic determinants responsible for tetracycline resistance in nine randomly chosen isolates revealed the presence of and/or This work confirms the occurrence and the continuous emergence of antibiotic-resistant strains in environmental samples; also, the presence of quinolone-resistant spp. in aquatic sources used for water supply and irrigation represents a potential risk for human health.
PubMed: 33802125
DOI: 10.3390/antibiotics10030288 -
Journal of Food Protection Jan 2021Arcobacter species are gram-negative rods that have been implicated in food- and waterborne illness. Although various cultural isolation methods have been proposed, the...
ABSTRACT
Arcobacter species are gram-negative rods that have been implicated in food- and waterborne illness. Although various cultural isolation methods have been proposed, the current procedures are unable to fully suppress the growth of background microbiota present in food samples, which inhibits Arcobacter isolation. The purpose of this study was to develop a selective enrichment broth and chromogenic plating medium to detect three Arcobacter species that have been recognized as emerging foodborne pathogens: Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The developed Nguyen-Restaino-Juárez (NRJ) Arcobacter detection system consists of a selective enrichment broth (NRJ-B) and a selective-differential plating medium (NRJ-M). The protocol of the detection method was determined by evaluating the growth of A. butzleri, A. cryaerophilus, and A. skirrowii under various temperatures (30, 35, and 42°C) and incubation (aerobic, microaerophilic, and anaerobic) conditions. Additionally, 47 Arcobacter strains and 39 non-Arcobacter strains were tested in inclusivity and exclusivity evaluations of NRJ-B and NRJ-M. Overall, the study determined that the optimal growth conditions of Arcobacter species using the NRJ Arcobacter detection system were aerobic incubation at 30°C. NRJ-B supported good growth of A. butzleri, A. cryaerophilus, and A. skirrowii while effectively suppressing the growth of non-Arcobacter strains after 48 h. Furthermore, NRJ-M yielded 97.8% inclusivity and 100.0% exclusivity using the tested strains and resulted in salmon-pigmented Arcobacter colonies (1.0 to 1.5 mm in diameter) after 72 h. The novel protocol is the first to develop a chromogenic plating medium for the isolation of Arcobacter species. This simple and accurate test method would greatly contribute to understanding the distribution of pathogenic Arcobacter species in food samples.
Topics: Agar; Arcobacter; Culture Media
PubMed: 33411930
DOI: 10.4315/JFP-20-245 -
Microbial Pathogenesis Mar 2021Arcobacter spp colonize in human and animals intestine and cause food-associated infections. Hence, characterization of their virulence potential and health impacts is...
Arcobacter spp colonize in human and animals intestine and cause food-associated infections. Hence, characterization of their virulence potential and health impacts is required. Our subject was isolation and characterization of Arcobacter spp, from meat marketplaces. A total of 1297 fresh raw cattle meat samples were purchased randomly from various marketplaces in Baghdad, Iraq. One-hundred and twenty isolates were identified, including Arcobacter butzleri (A. butzleri n = 100) and Arcobacter cryaerophilus (A. cryaerophilus n = 20). Susceptibility to antimicrobials was examined using Kirby-Bauer disc diffusion method. Molecular investigation of antibiotic resistance and virulence factors was also conducted using polymerase chain reaction (PCR) technique. Most of A. butzleri were resistant to tetracycline (72%), amoxicillin (69%), erythromycin (67%) and cefoxitin (66%), while 33% and 6% of them were resistant to ceftazidime and carbapenems, respectively. All were susceptible to gentamicin, colistin and fosfomycin. Fifty-five and nine isolates of A. butzleri and A. cryaerophilus were respectively multidrug-resistant (MDR). The existence of tetA, tetB, dfrA, sul1, bla and bla included 61%, 58%, 57%, 34%, 46% and 3%, respectively. The virulence genes cadF, irgA, tylA, cdtC and cdtA genes were detected in all the A. butzleri and A. cryaerophilus isolates. While, ciaB mviN and pldA genes were respectively detected in 91%, 88% and 84% of A. butzleri and 97%, 93% and 87% of A. cryaerophilus isolates. There was a significant relation between MDR and existence of virulence genes. Existence of pathogenic and drug-resistant- Arcobacter spp in raw meat is a threat for human health, necessitating confirmation of quality and safety of meat products.
Topics: Animals; Arcobacter; Cattle; Food Microbiology; Iraq; Meat; Meat Products; Virulence Factors
PubMed: 33249163
DOI: 10.1016/j.micpath.2020.104649 -
Letters in Applied Microbiology Feb 2021This research aims to investigate the presence and pathogenic potential of Arcobacter in poultry meat samples purchased in the retail market of Valdivia (South of Chile)...
This research aims to investigate the presence and pathogenic potential of Arcobacter in poultry meat samples purchased in the retail market of Valdivia (South of Chile) as well as in faecal samples from backyard chickens from rural areas around this city. The isolates obtained were identified by molecular methods. Furthermore, putative virulence genes were assessed by PCR and the antimicrobial resistance was tested by phenotypic methods. Arcobacter was present in 41·6% of the samples, with the highest value in retail poultry meat (55·7%) followed by backyard production (28·0%). Arcobacter butzleri was the most prevalent species (75·6%) followed by Arcobacter skirrowii (14·8%) and Arcobacter cryaerophilus (9·6%). An 8·5% of A. butzleri strains from meat were resistant to both ciprofloxacin and tetracycline and 6·1% were resistant to erythromycin, while none was resistant to gentamycin, unlike strains from domestic chickens, which showed no resistance. Furthermore, A. butzleri strains from chicken meat presented a higher prevalence of virulence genes than strains from domestic chickens. In fact, in this last group, some genes (hecA, hecB and irgA) were completely absent. Therefore, this study provides insight on the epidemiology of Arcobacter in Chilean poultry and suggests that under traditional breeding conditions strains are, apparently, less pathogenic and drug resistant.
Topics: Animals; Anti-Bacterial Agents; Arcobacter; Chickens; Chile; Ciprofloxacin; Drug Resistance, Bacterial; Erythromycin; Feces; Food Microbiology; Gentamicins; Gram-Negative Bacterial Infections; Meat; Polymerase Chain Reaction; Poultry; Prevalence; Tetracycline; Virulence; Virulence Factors
PubMed: 33025583
DOI: 10.1111/lam.13404 -
Frontiers in Microbiology 2020(formerly ) is a globally emerging foodborne and zoonotic pathogen. However, little is known about the species' genomic features and diversity, antibiotic resistance...
(formerly ) is a globally emerging foodborne and zoonotic pathogen. However, little is known about the species' genomic features and diversity, antibiotic resistance and virulence. In this study, 27 strains from water poultry in Thuringia, Germany, were investigated using whole-genome sequencing. Four of these strains were sequenced using long- and short-read sequencing methods to obtain circularized genomes. The German strains belong to the cluster I. Cluster I genomes exhibited a high degree of genetic diversity in which variable sites comprised 9.1% of the core genome. The German strains formed three subgroups that contained 2, 6, and 9 strains, respectively. The genomic analysis of cluster I revealed variable presence of mobile elements and that 65% of the strains lack CRISPR systems. The four circularized genomes carried a ∼2 Mbp chromosome and a single megaplasmid (size 98.1-154.5 Kbp). The chromosome was densely packed with coding sequences (∼92%) and showed inversions and shifts in the gene blocks between different strains. Antimicrobial resistance was assessed using a gradient strip diffusion method and showed that all 27 strains were resistant to cefotaxime and susceptible to erythromycin, gentamicin, and ampicillin. Sixteen strains were also resistant to ciprofloxacin, whereas 23 were resistant to streptomycin. The genetic prediction of antibiotic resistance identified numerous efflux pumps similar to those found in . All strains harbored two beta-lactamase genes which may explain the cefotaxime resistance. A correlation between the A point mutation (Thr-85-Ile) and ciprofloxacin resistance was partially discovered in 15 out of 16 strains. virulence profiling showed a wide range of virulence factors including a full chemotaxis system and most of the flagellar genes. In contrast to , no urease cluster was found. This study provides new insights into the genomic variability of strains of cluster I. The different genetic makeup of these strains may contribute to the virulence of strains and the severity of the infections in humans.
PubMed: 32754133
DOI: 10.3389/fmicb.2020.01549 -
PLoS Neglected Tropical Diseases Jul 2020Leptospirosis is one of the most significant zoonoses across the world not only because of its impact on human and animal health but also because of the economic and...
BACKGROUND
Leptospirosis is one of the most significant zoonoses across the world not only because of its impact on human and animal health but also because of the economic and social impact on agrarian communities. Leptospirosis is endemic in Sri Lanka where paddy farming activities, the use of draught animals in agriculture, and peridomestic animals in urban and rural areas play important roles in maintaining the infection cycle of pathogenic Leptospira, especially concerning animals as a potential reservoir. In this study, an environmental DNA (eDNA) metabarcoding methodology was applied in two different agro-ecological regions of Sri Lanka to understand the eco-epidemiology of leptospirosis.
METHODOLOGY/PRINCIPAL FINDINGS
Irrigation water samples were collected in Kandy District (wet zone mid-country region 2) and Girandurukotte, Badulla District (intermediate zone low-country region 2); and analysed for the presence of pathogenic Leptospira, associated microbiome and the potential reservoir animals. Briefly, we generated PCR products for high-throughput sequencing of multiple amplicons through next-generation sequencing. The analysis of eDNA showed different environmental microbiomes in both regions and a higher diversity of Leptospira species circulating in Kandy than in Girandurukotte. Moreover, the number of sequence reads of pathogenic Leptospira species associated with clinical cases such as L. interrogans was higher in Kandy than in Girandurukotte. Kandy also showed more animal species associated with pathogenic bacterial species than Girandurukotte. Finally, several pathogenic bacterial species including Arcobacter cryaerophilus, responsible for abortion in animals, was shown to be associated with pathogenic Leptospira.
CONCLUSIONS/SIGNIFICANCE
Leptospirosis has been considered to be endemic in wet regions, consistently, leptospiral sequences were detected strongly in Kandy. The great Leptospira species diversity in Kandy observed in this study shows that the etiological agents of leptospirosis in Sri Lanka might be underestimated. Furthermore, our eDNA metabarcoding can be used to discriminate bacterial and animal species diversity in different regions and to explore environmental microbiomes to identify other associated bacterial pathogens in the environment.
Topics: Agricultural Irrigation; Animals; DNA, Bacterial; DNA, Environmental; Fresh Water; Humans; Leptospira; Leptospirosis; Phylogeny; Sri Lanka; Zoonoses
PubMed: 32701971
DOI: 10.1371/journal.pntd.0008437 -
Journal of Food Protection Dec 2020Arcobacter is considered an emergent foodborne enteropathogen. Despite the high prevalence of this genus in poultry, the occurrence of Arcobacter spp. contamination in...
ABSTRACT
Arcobacter is considered an emergent foodborne enteropathogen. Despite the high prevalence of this genus in poultry, the occurrence of Arcobacter spp. contamination in Tunisia remains unclear. The objectives of this study were (i) to isolate Arcobacter species (A. butzleri and A. cryaerophilus) by the culture method from different species of raw poultry meat, (ii) to verify the isolates by multiplex PCR (m-PCR) assay and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and (iii) to determine the antibiotic resistance profiles of the isolates. A total of 250 poultry product samples (149 chicken and 101 turkey) were collected from various supermarkets in Sfax. The samples consisted of breasts, wings, legs, and neck skins. The overall isolation frequency of Arcobacter spp. was 10.4%. Arcobacter spp. were found in 13.42% of the chicken samples and in 5.49% of the turkey samples. All the acquired isolates were subject to detailed confirmation with subsequent species classification using m-PCR and MALDI-TOF MS. A. butzleri was found in 22 samples (84.61%) and A. cryaerophilus in 4 samples (15.38%). Thus, m-PCR and MALDI-TOF MS were able to detect A. butzleri significantly better than the conventional method (χ2 = 49.1 and P < 0.001). Arcobacter was isolated from poultry in every season, at contamination levels of 30.76, 23.07, 19.23, and 26.92% in summer, spring, autumn, and winter, respectively. The disk diffusion method was used to determine the susceptibility of Arcobacter isolates to six antimicrobial drugs. All A. butzleri isolates (n = 24) were significantly resistant to erythromycin (P = 0.0015), ampicillin (P = 0.001), and ciprofloxacin (P = 0.05). All tested A. cryaerophilus strains (n = 4) were susceptible to ampicillin, gentamicin, and amoxicillin-clavulanic acid. Multidrug resistance was observed in 83% of the Arcobacter spp. isolates. Our study detected Arcobacter spp. in Tunisian poultry; because of their multidrug resistance, these species may constitute a public health problem.
Topics: Animals; Arcobacter; Drug Resistance, Microbial; Food Microbiology; Poultry; Tunisia
PubMed: 32634222
DOI: 10.4315/JFP-20-056 -
Applied and Environmental Microbiology Apr 2020Pathogenic bacteria in wastewater are generally considered to be efficiently removed in biological wastewater treatment plants. This understanding is almost solely based...
Pathogenic bacteria in wastewater are generally considered to be efficiently removed in biological wastewater treatment plants. This understanding is almost solely based on culture-based control measures, and here we show, by applying culture-independent methods, that the removal of species in the genus was less effective than for many other abundant genera in the influent wastewater. was one of the most abundant genera in influent wastewater at 14 municipal wastewater treatment plants and was also abundant in the "clean" effluent from all the plants, reaching up to 30% of all bacteria as analyzed by 16S rRNA gene amplicon sequencing. Metagenomic analyses, culturing, genome sequencing of isolates, and visualization by fluorescent hybridization (FISH) confirmed the presence of the human-pathogenic and in both influent and effluent. The main reason for the high relative abundance in the effluent was probably that cells, compared to those of other abundant genera in the influent, did not flocculate and attach well to the activated sludge flocs, leaving a relatively large fraction dispersed in the water phase. The study shows there is an urgent need for new standardized culture-independent measurements of pathogens in effluent wastewaters, e.g., amplicon sequencing, and an investigation of the problem on a global scale to quantify the risk for humans and livestock. The genus was unexpectedly abundant in the effluent from 14 Danish wastewater treatment plants treating municipal wastewater, and the species included the human-pathogenic and Recent studies have shown that is common in wastewater worldwide, so the study indicates that discharge of members of the genus may be a global problem, and further studies are needed to quantify the risk and potentially minimize the discharge. The study also shows that culture-based analyses are insufficient for proper effluent quality control, and new standardized culture-independent measurements of effluent quality encompassing most pathogens should be considered.
Topics: Arcobacter; Denmark; RNA, Bacterial; RNA, Ribosomal, 16S; Waste Disposal, Fluid; Wastewater
PubMed: 32111585
DOI: 10.1128/AEM.03044-19