BACKGROUND: In Addis Ababa, Ethiopia, open ditches along innner roads in residential areas serve to convey domestic wastewater and rainwater away from residences. Contamination of drinking water by wastewater through faulty distribution lines could expose households to waterborne illnesses. This prompted the study to assess the microbiological safety of wastewater and drinking water in Addis Ababa, identify the pathogens therein, and determine their antibiotic resistance patterns.
UNASSIGNED: O1, mainly Hikojima serotype, was isolated from 23 wastewater and 16 drinking water samples. Similarly, 19 wastewater and 10 drinking water samples yielded Escherichia coli O157:H7. V. cholerae O1 were 100% resistant to the penicillins (Amoxacillin and Ampicillin), and 51-82% were resistant to the cephalosporins. About 44% of the V. cholerae O1 isolates in this study were Extended Spectrum Beta-Lactamase (ESBL) producers. Moreover, 26% were resistant to Meropenem. Peperacillin/Tazobactam was the only effective β-lactam antibiotic against V. cholerae O1. V. cholerae O1 isolates showed 37 different patterns of multiple resistance ranging from a minimum of three to a maximum of ten antimicrobials. Of the E. coli O157:H7 isolates, 71% were ESBL producers. About 96% were resistant to Ampicillin. Amikacin and Gentamicin were very effective against E. coli O157:H7 isolates. The isolates from wastewater and drinking water showed multiple antibiotic resistance against three to eight antibiotic drugs.
CONCLUSIONS: Open ditches for wastewater conveyance along innner roads in residence areas and underground faulty municipal water distribution lines could be possible sources for V. cholerae O1 and E. coli O157:H7 infections to surrounding households and for dissemination of multiple drug resistance in humans and, potentially, the environment.

The Gram-negative Elizabethkingia express multiple antibiotic resistance and cause severe opportunistic infections. Vancomycin is commonly used to treat Gram-positive infections and has also been used to treat Elizabethkingia infections, even though Gram-negative organisms possess a vancomycin permeability barrier. Elizabethkingia anophelis appeared relatively vancomycin-susceptible and challenge with this drug led to morphological changes indicating cell lysis. In stark contrast, vancomycin growth challenge revealed that E. anophelis populations refractory to vancomycin emerged. In addition, E. anophelis vancomycin-selected mutants arose at high frequencies and demonstrated elevated vancomycin resistance and reduced susceptibility to other antimicrobials. All mutants possessed a SNP in a gene (vsr1 = vancomycin-susceptibility regulator 1) encoding a PadR family transcriptional regulator located in the putative operon vsr1-ORF551, which is conserved in other Elizabethkingia spp as well. This is the first report linking a padR homologue (vsr1) to antimicrobial resistance in a Gram-negative organism. We provide evidence to support that vsr1 acts as a negative regulator of vsr1-ORF551 and that vsr1-ORF551 upregulation is observed in vancomycin-selected mutants. Vancomycin-selected mutants also demonstrated reduced cell length indicating that cell wall synthesis is affected. ORF551 is a membrane-spanning protein with a small phage shock protein conserved domain. We hypothesize that since vancomycin-resistance is a function of membrane permeability in Gram-negative organisms, it is likely that the antimicrobial resistance mechanism in the vancomycin-selected mutants involves altered drug permeability.

Pathogenic, antibiotic-resistant, and biofilm-forming bacteria can be transferred to humans through the consumption of contaminated seafood. The present study was carried out to determine antibiotic resistance profiles and virulence determinants in biofilm-forming Enterococcus faecium isolated from seafood in Bangladesh. A total of 150 seafood samples, including shrimp (n = 50), crabs (n = 25), and marine fish (n = 75), were screened using cultural, staining, biochemical, polymerase chain reaction (PCR), Congo red (CR), and disk diffusion (DD) assays. In PCR, E. faecium was detected in 27.3% (41/150; CI95% 20.8; 34.9) of samples, where marine fish (34.7%, CI95% 24.9; 45.9) had the highest prevalence (p < 0.05) compared to crabs (32%, CI95% 17.2; 51.6) and shrimp (14%, CI95% 7.0; 26.1). Thirty-two (78.1%, CI95% 63.3; 88.0) of the E. faecium isolates were determined to be biofilm formers in the CR test, where 43.9% (18/41, CI95% 29.9; 59.0) and 34.2% (14/41, CI95% 21.6; 49.5) of the isolates were strong and intermediate biofilm formers, respectively. In PCR, virulence genes, i.e., pil (100%), ace (92.7%), agg (68.3%), fsrA (65.9%), gelE (63.4%), sprE (53.7%), fsrB (51.2%), and fsrC (43.9%), were detected in E. faecium isolates. All the E. faecium isolates were phenotypically resistant to ≥3 antimicrobial categories and ≥3 antibiotics, including WHO-classified reserve antibiotics linezolid (70.7%) and fosfomycin (19.5%). Moreover, the multiple antibiotic resistance index ranged up to 0.8, showing resistance to ten antibiotics and eight antibiotic classes. In this study, the prevalence of virulence genes and antibiotic resistance was significantly greater (p < 0.05) in strong biofilm-forming E. faecium strains as compared to strains with intermediate and non-biofilm-forming abilities. As far as we know, this study, for the first time in Bangladesh, determined antibiotic resistance and detected virulence genes in biofilm-forming E. faecium isolated from seafood samples. The data from this study could play a significant role in evaluating potential health hazards linked to the ingestion of uncooked or minimally processed seafood.

UNASSIGNED: Avocados are typically sold in unsanitary conditions at the retail markets in Ecuador, which can raise the risk of microbial contamination. These microorganisms could exhibit multi-antibiotic resistance (MAR), being a serious threat concern to human health. In this study, we aimed to evaluate the microbiota and its antibiotic resistance profile in avocado Guatemalan fruits (Persea nubigena var. guatemalensis), at ripe stage: immature, firm light green (ready to eat in 4 days), peel (AFPE) and pulp (AFPU), and mature intense green (ready to eat) peel (AMPE) and pulp (AMPU), to gain baseline information on the prevalence of MAR bacteria.
UNASSIGNED: Culture-independent (16S rRNA metagenomics) and culture-dependent approach (to detect specific indicator microorganisms) were used. Moreover, antibiotic susceptibility of selected target indicator bacteria was assessed providing information about the antibiotic resistance (AR) among the groups.
UNASSIGNED: Based on 16S rRNA gene metagenomic analysis, over 99.78% of reads were classified as bacteria in all samples. Shannon diversity index varies from 1.22 to 2.22, with the highest bacterial population assigned to AFPE samples (1327 species). The highest microbial counts of indicator Staphylococcus spp. (STAPHY), Enterobacter spp. (ENT), and Listeria spp. (LIST), were detected in AMPE samples. Thirty percent of the selected STAPHYs, and 20.91% of Enterobacter (ENT) clones were resistant to various classes of antibiotics. The MAR index varies between 0.25 to 0.88 and was clone-, and fruit ripe stage-dependent.
UNASSIGNED: The results indicated that ready to eat avocados contained detectable levels of MAR bacteria, including methicillin resistant (MR)-STAPHY, which may act as a potential vector for the spread of antibiotic resistance. To achieve the increase of the production and marketing of Fuerte cultivar in Ecuador, it is vitally important to consider valuable strategies to protect the fruits at the early ripe stage in future. Thus, it is crucial to set up efficient control measures and develop coordinated strategies to guarantee the microbiological quality of the food.

The presence of Staphylococcus aureus, a normal human flora on cellphones of different professionals in Ile-Ife was investigated with a view to determining their antibiotic susceptibility profile and nature of resistance and virulence genes. One hundred swab samples were collected aseptically from mobile phones of various users based on their profession. Surfaces of the mobile phones were swabbed and the streak plate method was used to isolate colonies showing characteristic golden yellow on mannitol salt agar plates. These isolates were further identified using standard microbiological methods. The antibiotic susceptibility of the isolates was determined using Kirby-Bauer\'s disk diffusion technique. Molecular detection of nuc, mecA and pvl genes in some isolates was carried out by polymerase chain reaction technique. All the 36 isolates obtained in this study were 100% resistant to amoxicillin and augmentin; the isolates also displayed 55.6%, 44.4% and 41.7% resistance to ceftriazone, erythromycin and chloramphenicol, respectively. Based on resistance to oxacillin, prevalence of methicillin resistant Staphylococcus aureus (MRSA) was 11.1%. Only one S. aureus was positive for plasmid analysis. MecA gene was genetically confirmed in four (4) out of the 16 suspected phenotypic MRSA strains, nuc gene was confirmed in all 28 isolates investigated, while there was no pvl gene in the strains investigated. Mobile phones harbor multiple antibiotics resistant S. aureus, which are responsible for important diseases in humans and could be difficult to manage with antibiotics thereby posing serious health risks.

Antimicrobial resistance bacteria are nowadays ubiquitous. Its presence has been reported in almost every type of source, from water for agricultural and recreative use, water distribution pipes, and wastewater, to food, fomites, and clinical samples. Enterobacteriaceae, especially Escherichia coli, are not the exception, showing an increased resistance to several antibiotics, causing a global health and economic burden. Therefore, the monitoring of fecal microbiota is important because it is present in numerous reservoirs where gene transfer between commensal and virulent bacteria can take place, representing a potential source of resistant E. coli. In this work, antibiotic resistance profiles of 150 E. coli isolates from environmental, animal, and human samples, collected in three rural areas in Panama, were analyzed. A total of 116 isolates were resistant to at least one of the nine antibiotics tested. Remarkably, almost 100% of these exhibited resistance to tetracycline. Plasmid-associated tetA and tetB genes were detected in 42.86% of the isolates analyzed, tetA being the most prevalent. These results suggest that tetracycline resistance would be used as a convenient indicator of genetic horizontal transfer within a community.

Sugarcane bagasse (SB), as a major by-product of sugarcane, is one of the most abundant organic matter and characterized by cheap and easily available carbon source in Hainan Island, China. The objective of this study was to isolate tropical cellulolytic bacteria from Hainan Island and demonstrate their prospects of utilization of SB as a low-cost carbon source to greatly reduce the cost of aquaculture. A total of 97 cellulolytic marine bacteria were isolated, of which, 58 cellulolytic marine bacteria displayed the hydrolysis capacity (HC) of more than 1, while 28 cellulolytic marine bacteria displayed more than 2. Of the 28 tropical cellulolytic bacterial strains with HC more than 2, Microbulbifer sp. CFW-C18 and Vibrio sp. MW-M19 exhibited excellent SB decomposition in a small-scale laboratory simulation of shrimp aquaculture, up to 75.31 and 74.35%, respectively, and both of them were safe for shrimps. Meanwhile, both of CFW-C18 and MW-M19 besides displaying low multiple antibiotic resistance (MAR) index, also increased the C/N ratio (CFW-C18: C/N ratio of 14.34; MW-M19: C/N ratio of 14.75) of the small-scale laboratory simulation of shrimp aquaculture by decreasing the nitrogen content after a supplement of SB for 15 days. More importantly, CFW-C18 and MW-M19 displayed a relatively low MAR index, 0.47 and 0.1, respectively, especially MW-M19, with the lowest MAR index (0.1), which was resistant to only three antibiotics, streptomycin, amikacin, and levofloxacin, indicating that this strain was safe and non-drug resistance for further use. Overall, tropical cellulolytic bacteria isolated from Hainan Island, especially CFW-C18 and MW-M19, will provide the proficient candidates as probiotics for further construction of the recirculating aquaculture system based on the supplement of low-cost external carbon source-SB.

Salmonella genomic island 1 (SGI1), an integrative mobilisable element (IME), was first reported 20 years ago, in the multidrug resistant Salmonella Typhimurium DT104 clone. Since this first report, many variants and relatives have been found in Salmonella enterica and Proteus mirabilis. Thanks to whole genome sequencing, more and more complete sequences of SGI1-related elements (SGI1-REs) have been reported in these last few years among Gammaproteobacteria. Here, the genetic organisation and main features common to SGI1-REs are summarised to help to classify them. Their integrases belong to the tyrosine-recombinase family and target the 3\'-end of the trmE gene. They share the same genetic organisation (integrase and excisionase genes, replicase module, SgaCD-like transcriptional activator genes, traN, traG, mpsB/mpsA genes) and they harbour AcaCD binding sites promoting their excision, replication and mobilisation in presence of A/C plasmid. SGI1-REs are mosaic structures suggesting that recombination events occurred between them. Most of them harbour a multiple antibiotic resistance (MAR) region and the plasticity of their MAR region show that SGI1-REs play a key role in antibiotic resistance and might help multiple antibiotic resistant bacteria to adapt to their environment. This might explain the emergence of clones with SGI1-REs.

Numerous prevalence studies of Vibrio spp. infection in fish have been extensively reported worldwide, including Malaysia. Unfortunately, information on the prevalence of Vibrio spp. in groupers (Epinephelus spp.) is limited. In this study, groupers obtained from nine farms located at different geographical regions in Malaysia were sampled for the presence of pathogenic Vibrio spp. and their susceptibility profiles against seven antibiotics.
Out of 270 grouper samples, 195 (72%) were detected with the presence of Vibrio spp. Vibrio communis showed highest prevalence in grouper (28%), followed by V. parahaemolyticus (25%), V. alginolyticus (19%), V. vulnificus (14%), V. rotiferianus (3%), Vibrio sp. (3%), V. campbellii (2%), V. mytili (2%), V. furnissii (2%), V. harveyi (1%), V. tubiashii (1%), V. fluvialis (0.3%) and V. diabolicus (0.3%). Assessment on the antibiotic susceptibility profiles of the Vibrio spp. revealed that majority of the isolates were susceptible to tetracycline, streptomycin, erythromycin and bacitracin, but resistance to ampicillin, penicillin G and vancomycin. The mean MAR index of the Vibrio isolates was 0.51, with 85% of the isolates showed MAR index value of higher than 0.2. Results indicate that the Vibrio spp. were continuously exposed to antibiotics. Furthermore, the plasmid profiles of Vibrio spp. showed that 38.7% of the isolates harbored plasmid with molecular weight of more than 10 kb, while 61.3% were without plasmid. During curing process, Vibrio spp. lost their plasmid, but remained resistant to ampicillin, penicillin G, bacitracin and vancomycin while a few isolates remained resistant to erythromycin, streptomycin and tetracycline. The results suggested that the resistance to antibiotics in isolated Vibrio spp. might be due to chromosomal and plasmid borne.
This study demonstrates the prevalence of Vibrio spp. in groupers and the distribution of multidrug resistance strains that could be of concern to the farmers in Malaysia. In addition, data from this study can be further used in fish disease management plan.

Vibrio Parahaemolyticus infections caused by the pandemic clone have become a global public health issue. The pandemic clone includes over ten sequence types and 49 serotypes. Several markers such as toxRS/new, orf8 and genomic islands were considered specific for pandemic strains, but subsequent studies later confirmed a lack of specificity. Thus, identifying stable indicators for the pandemic clone is still an open question. In recent years, several environmental pandemic strains are growing, constituting a new threat to seafood safety and human health. Traditional methods show limited discrimination in studying the microevolution of pandemic strains. For example, multilocus sequence typing divides many pandemic strains into ST3 type, making it difficult to further distinguish the variability within ST3 strains from different contexts. When using a whole genome sequencing-based technique, strains including those with the same sequence type, could be well separated. Whole genome sequencing-based technology also played important roles in dissecting the evolution process and revealing the mechanism underlying rapid serotype conversion within pandemic strains. In addition, the emergence of multiple-antibiotic resistant pandemic strains needs attention. Altogether, we are facing many challenges posed by pandemic V. parahaemolyticus strains, which need to be resolved in future studies.