Potential Human Pathogens Research Articles

MOLECULAR IDENTIFICATION OF ROSEOMONAS MUCOSA AND DETERMINATION THE SOME OF ITS VIRULENCE FACTORS

This study was aimed to characterize Roseomonas mucosa isolate using molecular genetics and determination some bacterial virulence factors such as capsular, biofilm formation and antimicrobial susceptibility. By using Polymerase chain reaction (PCR) technique, the molecular characterization of the R. mucosa was done by using 16S rRNA gene sequence and phylogenetic analysis. Antimicrobial susceptibility test was carried out by Kirby-Bauer disk diffusion method. Capsular detection was examined using nigrosin pigment. While Congo red agar and microtiter plate using enzyme-linked immunosorbent assay (ELISA) reader methods are used to determined bacterial isolate capability to produce a biofilm. The molecular characterization shows the bacterium that isolated from catheter of a dialysis Iraqi patient is belong to Roseomonas mucosa. The isolated R. mucosa strain showed resistant to Amoxicillin, methicillin, carbapenemes, bacitracin and Sulfamethoxazole. While it was susceptible to tobramycin, levofloxacin, colisitin sulphate, doxycycline, azithromycin and cephalexin. The results show R. mucosa was encapsulated. Furthermore, it has the ability to form weak biofilm. The findings suggest that in spite of this genus seems to have an overall little pathogenic potential for humans, Roseomonas sp. in particular, R. mucosa may be substantial pathogens in individuals with underlying medical complications.

Bacterial communities shift and influence in an acid mine drainage treatment using barium carbonate disperse alkaline substrate system

Chemical passive treatment systems used to remediate acid mine drainage has been evaluated based mainly on the reactivity of the chemical alkaline reagents, overlooking the activity of the microorganisms that proliferate in these artificial ecosystems. In this study, the bacterial communities of a unique passive treatment system known as BDAS (Barium carbonate Dispersed Alkaline Substrate) were investigated using 16S rRNA gene metagenomic sequencing combined with hydrochemical characterization of the AMD and phenotypic characterization of biogenic precipitates. According to the hydrochemical characterization, the water quality improved as the water progressed through the system, with a drastic increase in the pH (up to alkaline conditions) and total organic carbon, as well as the removal of main contaminants such as Ca2+, SO42−, Fe3+, Al3+, and Mn2+. These environmental changes resulted in an increase in bacterial diversity (richness) after the inlet and in the shift of the bacterial communities from chemoautotrophs (e.g., Ferrovum and Acidiphilum) to chemoheterotrophs (e.g., Brevundimonas and Geobacter). Some of these taxa harbour potential to immobilize metals, aiding in the treatment of the water. One of the mechanisms involved in the immobilization of metals is microbially induced calcium carbonate precipitation, which seems to occur spontaneously in BDAS. The production of biofilm was also observed in most parts of the system, except in the inlet, helping with the removal of metals. However, in the long run, the build-up of biofilm and precipitation of metals could clog (i.e., biofouling) the pores of the matrix, reducing the treatment efficiency. Potential human pathogens (e.g. Legionella) were also detected in BDAS indicating the need for a treatment step at the end of the system to remove pathogenic microorganisms. These findings present a new perspective of the bacterial communities and their effects (both positively and negatively) in a chemical passive treatment system.

Increasing Antimicrobial Resistance and Potential Human Bacterial Pathogens in an Invasive Land Snail Driven by Urbanization

Our understanding of the role urbanization has in augmenting invasive species that carry human bacterial pathogens and antimicrobial resistance (AMR) remains poorly understood. Here, we investigated the gut bacterial communities, antibiotic resistance genes (ARGs) and potential antibiotic-resistant pathogens in giant African snails (Achatina fulica) collected across an urbanization gradient in Xiamen, China (n = 108). There was a lack of correlation between the microbial profiles of giant African snails and the soils of their habitats, and the resistome and human-associated bacteria were significantly higher than those of native snails as well as soils. We observed high diversity (601 ARG subtypes) and abundance (1.5 copies per 16S rRNA gene) of giant African snail gut resistome. Moreover, giant African snails in more urban areas had greater diversity and abundance of high-risk ARGs and potential human bacterial pathogens (e.g., ESKAPE pathogens). We highlight that urbanization significantly impacted the gut microbiomes and resistomes of these invasive snails, indicating that they harbor greater biological contaminants such as ARGs and potential human bacterial pathogens than native snails and soils. This study advances our understanding of the effect of urbanization on human bacterial pathogens and AMR in a problematic invasive snail and should help combat risks associated with invasive species under the One Health framework.

An updated review of diffuse mesothelioma of the pleura – A sentinel health event of potential elongate mineral particle pathogenicity

There are approximately 400 inorganic minerals in the Earth's crust, some of which can be encountered as elongate mineral particles [EMPs] with dimensional characteristics similar to the six minerals known as asbestos and other asbestiform amphiboles with established human pathogenicity. In addition, the rapidly developing field of nanotechnology is producing an ever-increasing array of high aspect ratio engineered nanomaterials [HARNs] with physical dimensions and biodurability similar to the asbestos fiber types with recognized pathogenic potential. Many of these non-asbestos/non-asbestiform EMPs and HARNs with the potential for aerosolization into the breathing zones of workers and in individuals in non-occupational environments have not yet been thoroughly studied with respect to their potential human pathogenicity, a fact which obviously poses concerns for both occupational health and public health professionals. On the basis of dose-response considerations it seems reasonable to infer that if any of these non-regulated EMPs or HARNs actually are pathogenic, then those mineral fiber exposure-induced disorders associated with the lowest cumulative exposure doses of the commercial amphibole types of asbestos, that is, diffuse mesothelioma of the pleura, and its non-malignant correlate of benign parietal pleural plaques, are those which are most likely to occur following inhalational exposures to any of the non-regulated EMPs and HARNs. Because of that observation, this paper reviews certain aspects of diffuse mesothelioma, including a summary of recent changes in the nomenclature of diffuse mesothelioma of the pleura; of both the descriptive and the analytical epidemiology of the disease; of the etiologies of mesothelioma, both “exposure” related and endogenous in nature; and of the asbestos population attributable fraction for diffuse mesotheliomas in the USA, both historically and in the future.

Gastrointestinal Polyparasitism in Bushmeat in Zadie Department in Northeast Gabon

Wild animals harbor pathogens that can be infectious agents for humans, including parasites. This study aimed to identify gastrointestinal parasites and assess their prevalence and the potential risk for humans associated with consuming these animals. The research was conducted from August to December 2019. Parasitological analyses were carried out on the feces and intestines of 113 wild animals, including antelopes (24), duikers (58), porcupines (18), small monkeys (Cercopithecus) (8), nandinia (2), pangolin (1), genet (1), and a crocodile (1), from the Zadié Department in the province of Ogooué-Ivindo in the northeast of Gabon. The results revealed 15 taxa of gastrointestinal parasites, including nine nematodes: Strongylids (61/113), Strongyloides spp. (21/113), Ascaris spp. (21/113), Trichuris spp. (39/113), Capillaria spp. (9/113), Protostrongylus spp. (5/113), Enterobius spp. (8/113), Toxocara spp. (7/113) and Mammomonogamus spp. (5/113); three species of protozoa, namely Balantidium spp. (12/113), Eimeria spp. (17/113), and Entamoeba spp. (9/113); two species of trematodes, namely Fasciola spp. (18/113) and Paramphistomum spp. (21/113); and cestode species, Taenia spp. (1/113). The prevalence of gastrointestinal parasitism in these animals was 85.84% (97/113). In addition, among these parasitic taxa, some are potential pathogens for humans, such as Ascaris spp., Balantidium spp., Entamoeba spp., and Taenia spp. The consumption of games, particularly offal, infested by these parasites, could threaten human health.

Emergence of multidrug-resistant Acinetobacter baumannii under fluctuating levofloxacin concentration and its control by chlorine and UV disinfection

Although wastewater treatment plants are considered hotspots for the spread of antibiotic-resistant bacteria (ARB) and antibiotic-resistant genes (ARGs), few studies have explored the mechanisms underlying the emergence of multidrug-resistance. Using four sequencing batch reactors under fluctuating concentrations of levofloxacin (LVX), we isolated 440 isolates, which were identified based on the sequence of the 16 S rRNA gene and growth at 37 ºC, focusing on potential human pathogens. We then evaluated the multidrug-resistance of 62% of these isolates by determining the minimum inhibition concentration (MIC) of erythromycin, tetracycline, chloramphenicol, and LVX in the emergence of ARB. The average MIC was increased more than 100-fold in the first exposure, whereas did not show the same response after re-exposure due to different LVX concentrations between the 2 conditions. We identified 33 Acinetobacter baumannii isolates that emerged only during re-exposure, especially in R-2 and R-3, in which the LVX concentration was changed from 4 and 16 mg/L, respectively, to 128 mg/L. Moreover, we assessed the disinfection efficiency of chlorine/UV treatment against pathogenic ARB. The growth of multidrug-resistant A. baumannii isolates was decreased by 0.9–7.4 log10 following disinfection treatment with > 0.5 mg /L Cl2 or > 30 min UV irradiation.

Antibiotic Resistance, Virulence Gene Detection, and Biofilm Formation in Aeromonas spp. Isolated from Fish and Humans in Egypt.

The genus Aeromonas is widely distributed in aquatic environments and is recognized as a potential human pathogen. Some Aeromonas species are able to cause a wide spectrum of diseases, mainly gastroenteritis, skin and soft-tissue infections, bacteremia, and sepsis. The aim of the current study was to determine the prevalence of Aeromonas spp. in raw fish markets and humans in Zagazig, Egypt; identify the factors that contribute to virulence; determine the isolates' profile of antibiotic resistance; and to elucidate the ability of Aeromonas spp. to form biofilms. The examined samples included fish tissues and organs from tilapia (Oreochromis niloticus, n = 160) and mugil (Mugil cephalus, n = 105), and human skin swabs (n = 51) and fecal samples (n = 27). Based on biochemical and PCR assays, 11 isolates (3.2%) were confirmed as Aeromonas spp. and four isolates (1.2%) were confirmed as A. hydrophila. The virulence genes including haemolysin (hyl A) and aerolysin (aer) were detected using PCR in A. hydrophila in percentages of 25% and 50%, respectively. The antimicrobial resistance of Aeromonas spp. was assessed against 14 antibiotics comprising six classes. The resistance to cefixime (81.8%) and tobramycin (45.4%) was observed. The multiple antibiotic resistance (MAR) index ranged between 0.142-0.642 with 64.2% of the isolates having MAR values equal to 0.642. Biofilm formation capacity was assessed using a microtiter plate assay, and two isolates (18.1%) were classified as biofilm producers. This study establishes a baseline for monitoring and controlling the multidrug-resistant Aeromonas spp. and especially A. hydrophila in marine foods consumed in our country to protect humans and animals.

The current state of the problem of tick-borne encephalitis in Russia and the world

Relevance. Despite the successes achieved over the 85-year history of the study of tick-borne encephalitis (TBE), there are still many controversial and unresolved issues. It is obvious that in the second decade of the XXI century, this natural focal neuroinfection, as before, poses a great threat to public health not only in Russia, but also in endemic countries of the world.Aim. To present modern aspects of etiology, epidemiology, diagnosis, specific prevention of TBE in Russia and endemic countries of the world.Conclusions. Currently, there are 4 genotypes of the TBE virus: Far Eastern, European, Siberian and Baikal, each of which has its own area, pathogenic potential for humans. Nosoareal of TBE covers most of the territory of Russia, 29 European countries. Cases of TBE disease or viral activity are registered in six Asian countries. For the correct verification of the diagnosis, criteria for the diagnosis of cases of the disease have been developed, which mainly concern unvaccinated persons. Since highly effective etiotropic drugs for the treatment of TBE have not yet been developed, the only strategy for reducing the incidence of TBE is mass vaccination of the population of endemic regions. Due to the increase in the rate of vaccination in Russia and European countries, in order to improve epidemiological surveillance of TBE, standards for specific laboratory diagnosis of the disease among vaccinated persons, clarification of the causes of the incidence of vaccinated, as well as deaths among them, need to be clarified. In conditions of combination of natural foci of TBE and other tick-borne infections, it is important to develop diagnostic algorithms, including differential diagnosis of this disease with other infections transmitted by Ixodic ticks.

Microbial Risk Assessment of Mature Compost from Human Excreta, Cattle Manure, Organic Waste, and Biochar

Lack of sanitation is the underlying cause of many diarrheal infections and associated deaths. Improving sanitation through the set-up of ecological sanitation dry toilets, followed by the thermophilic composting of human excreta, could offer a solution. In addition, treating the excreta via thermophilic composting allows us to recycle the nutrients to be used as fertilizer for agriculture. However, for this purpose, the compost should be free of pathogens. We conducted a thermophilic composting trial over 204 to 256 days with human excreta, along with vegetable scraps and teff straw, with and without biochar. A sawdust–cattle manure mixture with the same supplements served as a control treatment. To evaluate the hygienic quality of the mature compost, the bacterial indicators Escherichia coli and Salmonella were assessed using the cultivation-based most probable number method. In addition, Ascaris lumbricoides eggs were quantified through light microscopy. The amount of detected E. coli was below the thresholds of German and European regulations for organic fertilizer. Salmonella and Ascaris eggs were not detected. No significant differences between the treatments were observed. Thus, the composting process was efficient in decreasing the number of potential human pathogens. The mature compost fulfilled the legal regulations on organic fertilizer regarding potential human pathogens.

Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes

The development and spread of antibiotic resistance (AR) through microbiota associated with freshwater bodies is a major global health concern. In the present study, freshwater samples were collected and analyzed with respect to the total bacterial diversity and AR genes (ARGs) using both conventional culture-based techniques and a high-throughput culture-independent metagenomic approach. This paper presents a systematic protocol for the enumeration of the total and antibiotic-resistant culturable bacteria from freshwater samples and the determination of phenotypic and genotypic resistance in the culturable isolates. Further, we report the use of whole metagenomic analysis of the total metagenomic DNA extracted from the freshwater sample for the identification of the overall bacterial diversity, including non-culturable bacteria, and the identification of the total pool of different ARGs (resistome) in the water body. Following these detailed protocols, we observed a high antibiotic-resistant bacteria load in the range of 9.6 × 105-1.2 × 109 CFU/mL. Most isolates were resistant to the multiple tested antibiotics, including cefotaxime, ampicillin, levofloxacin, chloramphenicol, ceftriaxone, gentamicin, neomycin, trimethoprim, and ciprofloxacin, with multiple antibiotic resistance (MAR) indexes of ≥0.2, indicating high levels of resistance in the isolates. The 16S rRNA sequencing identified potential human pathogens, such as Klebsiella pneumoniae,and opportunistic bacteria, such as Comamonas spp., Micrococcus spp., Arthrobacter spp., and Aeromonas spp. The molecular characterization of the isolates showed the presence of various ARGs, such as blaTEM, blaCTX-M (β-lactams), aadA, aac (6')-Ib (aminoglycosides), and dfr1 (trimethoprims), which was also confirmed by the whole metagenomic DNA analysis. A high prevalence of other ARGs encoding for antibiotic efflux pumps-mtrA, macB, mdtA, acrD, β-lactamases-SMB-1, VIM-20, ccrA, ampC, blaZ, thechloramphenicol acetyltransferase gene catB10, andthe rifampicin resistance gene rphB-was also detected in the metagenomic DNA. With the help of the protocols discussed in this study, we confirmed the presence of waterborne MAR bacteria with diverse AR phenotypic and genotypic traits. Thus, whole metagenomic DNA analysis can be used as a complementary technique to conventional culture-based techniques to determine the overall AR status of a water body.

Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes

The development and spread of antibiotic resistance (AR) through microbiota associated with freshwater bodies is a major global health concern. In the present study, freshwater samples were collected and analyzed with respect to the total bacterial diversity and AR genes (ARGs) using both conventional culture-based techniques and a high-throughput culture-independent metagenomic approach. This paper presents a systematic protocol for the enumeration of the total and antibiotic-resistant culturable bacteria from freshwater samples and the determination of phenotypic and genotypic resistance in the culturable isolates. Further, we report the use of whole metagenomic analysis of the total metagenomic DNA extracted from the freshwater sample for the identification of the overall bacterial diversity, including non-culturable bacteria, and the identification of the total pool of different ARGs (resistome) in the water body. Following these detailed protocols, we observed a high antibiotic-resistant bacteria load in the range of 9.6 × 105-1.2 × 109 CFU/mL. Most isolates were resistant to the multiple tested antibiotics, including cefotaxime, ampicillin, levofloxacin, chloramphenicol, ceftriaxone, gentamicin, neomycin, trimethoprim, and ciprofloxacin, with multiple antibiotic resistance (MAR) indexes of ≥0.2, indicating high levels of resistance in the isolates. The 16S rRNA sequencing identified potential human pathogens, such as Klebsiella pneumoniae, and opportunistic bacteria, such as Comamonas spp., Micrococcus spp., Arthrobacter spp., and Aeromonas spp. The molecular characterization of the isolates showed the presence of various ARGs, such as blaTEM, blaCTX-M (β-lactams), aadA, aac (6')-Ib (aminoglycosides), and dfr1 (trimethoprims), which was also confirmed by the whole metagenomic DNA analysis. A high prevalence of other ARGs encoding for antibiotic efflux pumps-mtrA, macB, mdtA, acrD, β-lactamases-SMB-1, VIM-20, ccrA, ampC, blaZ, the chloramphenicol acetyltransferase gene catB10, and the rifampicin resistance gene rphB-was also detected in the metagenomic DNA. With the help of the protocols discussed in this study, we confirmed the presence of waterborne MAR bacteria with diverse AR phenotypic and genotypic traits. Thus, whole metagenomic DNA analysis can be used as a complementary technique to conventional culture-based techniques to determine the overall AR status of a water body.

In vitro digestion of ESC-resistant Escherichia coli from poultry meat and evaluation of human health risk.

The spread of antimicrobial resistance (AMR) has become a threat against human and animal health. Third and fourth generation cephalosporins have been defined as critically important antimicrobials by The World Health Organization. Exposure to Extended spectrum cephalosporin-resistant E. coli may result in consumers becoming carriers if these bacteria colonize the human gut or their resistance genes spread to other bacteria in the gut microbiota. In the case that these resistant bacteria at later occasions cause disease, their resistance characteristics may lead to failure of treatment and increased mortality. We hypothesized that ESC-resistant E. coli from poultry can survive digestion and thereby cause infections and/or spread their respective resistance traits within the gastro-intestinal tract. In this study, a selection of 31 ESC-resistant E. coli isolates from retail chicken meat was exposed to a static in vitro digestion model (INFOGEST). Their survival, alteration of colonizing characteristics in addition to conjugational abilities were investigated before and after digestion. Whole genome data from all isolates were screened through a custom-made virulence database of over 1100 genes for virulence- and colonizing factors. All isolates were able to survive digestion. Most of the isolates (24/31) were able to transfer their bla CMY2-containing plasmid to E. coli DH5-á, with a general decline in conjugation frequency of digested isolates compared to non-digested. Overall, the isolates showed a higher degree of cell adhesion than cell invasion, with a slight increase after digestion compared non-digested, except for three isolates that displayed a major increase of invasion. These isolates also harbored genes facilitating invasion. In the virulence-associated gene analysis two isolates were categorized as UPEC, and one isolate was considered a hybrid pathogen. Altogether the pathogenic potential of these isolates is highly dependent on the individual isolate and its characteristics. Poultry meat may represent a reservoir and be a vehicle for dissemination of potential human pathogens and resistance determinants, and the ESC-resistance may complicate treatment in the case of an infection.

A mouth watering case of Mycoplasma salivarium native mitral valve endocarditis: ‘case report’

BackgroundMycoplasma salivarium is part of our commensal oral flora and readily resides in dental plaque. Although considered indolent, few case reports have documented its pathogenic potential in humans. To this day no case of Mycoplasma salivarium infectious endocarditis has ever been described.Case presentationOur report describes a challenging case of Mycoplasma salivarium endocarditis, with a patient presenting with oligoarticular joint swelling, and later on in the course of his disease developed signs of right-sided heart failure. The diagnosis was initially mistaken for septic gonarthritis and was later established on the basis of echocardiography and eubacterial PCR of joint fluid.ConclusionThis report describes a first documented case of Mycoplasma salivarium culture negative endocarditis that was successfully treated with targeted antimicrobial therapy. Specific antimicrobial therapy targeting Mycoplasma spp, lead to clinical improvement, with radiological regression of the lesion and the resolution of the serum inflammation biomarkers.

West Nile virus in India: An update on its genetic lineages.

West Nile virus (WNV) is a rapidly spreading mosquito-transmitted zoonotic flavivirus. Mosquitoes belonging to the genus Culex are incriminated as the principal vectors of the virus, which causes West Nile fever (WNF) in humans. Manifestations of WNF include a mild, self-limiting, flu-like illness, which in severe cases (rare) may progress to encephalitis, resulting in life-threatening consequences. WNV is geographically distributed worldwide, covering Africa, the Americas, Europe, and Asia (except Antarctica). The virus exists in a bird-mosquito transmission cycle in nature, with humans and horses as incidental/accidental hosts. The virus can infect a large variety of hosts worldwide, i.e., about 300 birds and around 70 different mosquito species belonging to several genera. For a long time, it was believed that WNV was not highly virulent and caused only mild infection globally. However, the recent frequent and increasing incidence of clinically severe WNV infections, such as encephalitis in humans and horses with significant mortality, has been reported in the Americas, Europe, and several East Asian countries. The emergence of lineage 2 strains endemic to Africa, with epidemic potential in humans and horses in Europe, is considered a serious global health concern. Although WNV is known to circulate in India since 1952, its re-emergence with severe neuro-invasive pathogenic potential in humans in Assam, Kerala, West Bengal and Tamil Nadu states signals urgent efforts to understand the dynamics of circulating strains with regard to its vector, hosts, and environment. This could be done by prioritizing "One Health" approach for developing effective preventive and control strategies. In view of the global interest, we present an overview of the circulating genetic lineages of WNV in India in comparison with the global scenario. In addition, we stress on holistic approaches of "One Health" strategy as the current need of the hour for designing effective preventive and control strategies in the country.

Comunidades bacterianas em queijos artesanais de leite bovino cru da região sul do Brasil

ABSTRACT: The production of artisanal cheeses made with raw bovine milk has grown in the southern region of Brazil. It is important to obtain information about the risks of this practice, especially concerning food safety. In this study, next-generation sequencing was used to identify and characterize the bacterial communities of artisanal raw milk cheeses. We analyzed one pool of five raw milk samples (control group M1) from different dairy farms and nine pools (M2-M10) of 45 artisanal raw milk cheeses.The characterization of the bacterial communities included 199 species distributed across 59 different genera dispersed among the samples. Among the genera observed, 11 were classified as beneficial to the aroma, flavour, colour, and texture of the cheese. Thirty-one genera were classified as harmful to these characteristics. Another 17 were classified as potential pathogens for animals and humans, including Aeromonas, Bacillus, Cronobacter, Salmonella, Staphylococcus, and bacteria of the coliform group, including E. coli and Klebsiella. There was a significant difference (P < 0.05) in the number of bacterial communities identified between the control group (M1) and the two pools of artisanal raw milk cheeses (M2 and M8). This study demonstrated that next-generation sequencing provides in-depth information on the composition of the microbiota in artisanal raw milk cheeses, characterizing bacterial communities, identifying the wide microbial diversity, and identifying microbial benefits and risks.

Household spices and minerals as alternative disinfectants for mobile phones

Mobile phones play an essential role in enhancing personal, social, and professional life in many ways, including to access to communication, information, and entertainment. However, mobile phones also come in contact with environments containing high densities of microorganisms, such as hospitals, schools, restrooms, and kitchens, suggesting that mobile phones could also act as carriers of disease. Commercial disinfectants are highly effective against potential human pathogens found on mobile phones, yet these disinfectants also contain chemicals that are toxic to both human health and the environment. In addition, there is evidence that modern microorganisms are becoming resistant to commercially available disinfectants. We hypothesized that household spices and minerals, such as garlic, mint, zinc, and iron, could act as alternate solutions for disinfecting mobile phones after daily use. Our results demonstrate that garlic, iron, and zinc are just as effective at suppressing microorganisms compared to commercially available cleaners, Clorox and Pine-Sol. These results could help guide further research aimed at producing more environmentally friendly cleaners and reducing the need for toxic chemicals in household disinfectants.

First report on the molecular characteristics of mcr-1 colistin resistant E. coli isolated from retail broiler meat in Bangladesh

Poultry meat is considered as a potential source of colistin resistant Escherichia coli (CREC). This study aimed to determine the prevalence and characteristics of CREC in broiler meat and ascertain their possible zoonotic potential(s). Broiler meat (n = 104) comprising 26 of each of the thigh, breast, liver, and proventriculus-gizzard was purchased from the retail outlets, Bangladesh. CREC was isolated from the meat samples on MacConkey agar plates containing colistin sulfate followed by PCR confirmation, mcr subtyping (mcr-1 to mcr-5), phylogenetic grouping and detailed molecular characterization through whole genome sequencing (WGS). Antimicrobial resistance of the CREC isolates were evaluated by disc diffusion method and MIC (minimum inhibitory concentration) of colistin sulfate was determined by broth microdilution. The investigation revealed 58 (55.77 %) of 104 samples as positive for CREC, and 53 (91.38 %) of CREC isolates carried mcr-1 gene with no other mcr subtypes evident. Most of the CREC belonged to commensal E. coli (66.04 %) with some pathogenic phylotypes (33.96 %) based on dichotomous decision tree. All the mcr-1 CREC isolates were multidrug-resistant (MDR) and had MICs of 4–8 μg/mL colistin sulfate. WGS of a commensal MDR mcr-1 CREC strain 1ChBEc2mcr revealed as a potential human pathogen belonging to ST162 that harbored 60 virulence factors associated genes (VFGs). The mcr-1 gene in 1ChBEc2mcr genome was located on a plasmid (p1ChBEc2mcr) and showed nucleotide similarities (>95 %) to another plasmid reported from human E. coli in Bangladesh. Beyond mcr-1 gene, this plasmid (p1ChBEc2mcr) also harbored genes related to aminoglycoside, beta-lactams, macrolides, and tetracycline resistance. Presence of similar mcr-1 carrying plasmids in broiler and human CREC denotes a threat of possibly human to avian (broiler) or vice-versa transfer of mcr-1 CREC through close contact as prevailing in the retail outlets of Bangladesh.

Manure and biochar have limited effect on lettuce leaf endophyte resistome

Soil amendment with manure compost and biochar is widely adopted to improve soil fertility and promote plant growth, and their effects on soil microbial communities and resistome have been well documented. However, there is sparse information regarding their effects on vegetable endophytes, which represent a major source of human exposure to pathogens and antibiotic resistance genes (ARGs) when eaten raw. Here, we investigated the impacts of manure compost or biochar addition on the bacterial community compositions and ARGs in the soil-lettuce continuum including soil, seed, leaf, and root samples. A total of 137 ARGs and 31 mobile genetic elements (MGEs) were detected in all the samples after 60 days of cultivation. The relative abundance of ARGs and the diversity of bacteria communities presented a consistent decreasing trend from soil to root endophytes, then leaf endophytes. Manure compost addition increased the diversity and abundance of ARGs in soil, while significant changes in the ARG profiles and bacterial communities were not observed in leaf endophytes after manure compost or biochar addition, or both. Bipartite networks analysis suggested that seed microbiome was one of the major sources of plant endophytes and ARGs. Twenty potential human pathogens were isolated from lettuce, indicating potential exposure risk to pathogens via the consumption of raw lettuce. These results suggest limited impacts of manure compost and biochar addition on lettuce endophytes and highlight the contribution of seed microbiome to endophyte ARG profiles.

Potential Pathogenicity of Aeromonas spp. Recovered in River Water, Soil, and Vegetation from a Natural Recreational Area.

The genus Aeromonas is widely distributed in aquatic environments and is recognized as a potential human pathogen. Some Aeromonas species are able to cause a wide spectrum of diseases, mainly gastroenteritis, skin and soft-tissue infections, bacteremia, and sepsis. Currently, untreated river water is used for irrigation and recreational purposes. In this study, the Aeromonas spp. present in a river recreational environment was investigated by quantifying its presence in water, soil, and vegetation using three techniques: qPCR, plate counting in selective ADA medium, and Most Probable Number, in parallel. The presence of clones in the three types of samples was elucidated through genotyping with the ERIC-PCR technique, whereas the identification of the isolated Aeromonas was carried out by sequencing the rpoD gene. Finally, the pathogenic potential of some of the strains was explored by studying the presence and expression of virulence genes characteristic of the genus, their antimicrobial susceptibility profile, as well as the quantification of their cell damage and intracellular survival in an in vitro macrophages infection model. The results showed the presence of Aeromonas in all samples with the three quantification methods, with Aeromonas popoffii being the most prevalent species. The presence of strains with the same genotype (ERIC-PCR) was also confirmed in different samples. Some of the strains showed a high level of cell damage and intracellular bacterial survival, as well as the presence of various virulence factors. Furthermore, these strains showed resistance to some of the antibiotics tested and used therapeutically in both humans and animals. These results indicate that the presence of Aeromonas in this environment may represent a biosanitary risk that could be a public health problem.

Development of nine microsatellite loci for Trypanosoma lewisi, a potential human pathogen in Western Africa and South-East Asia, and preliminary population genetics analyses

Development of nine microsatellite loci for Trypanosoma lewisi, a potential human pathogen in Western Africa and South-East Asia, and preliminary population genetics analyses