Resistomes Associated with Transmissible Bacterial Pathogens in Water and Sediments from Oko-Agbon Stream, Makoko, Lagos, Nigeria

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Apeh, Emmanuel A. Meshach, Oluwatosin T. Fagbemi, Modupeade C. Adetunji, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6341991/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 03 Feb, 2026 Read the published version in BMC Microbiology → Version 1 posted 13 You are reading this latest preprint version Abstract Background: Water is essential for life; however, it serves as the route of transmission for many infectious pathogens. The increasing prevalence of multidrug-resistant (MDR) bacteria in water samples poses a significant public health risk. Aim : This study aimed to characterize and analyze the molecular profile of selected MDR bacteria isolated from water and sediment samples from the Oko-Agbon stream in the Makoko community. Method : The bacteria were identified using conventional methods and subsequently subjected to antimicrobial susceptibility testing using Kirby-Bauer disk diffusion technique. The MDR strains were screened for ESBLs and Carbapenemase genes phenotypically by double disk synergy method and genotypically by PCR amplification. The specific genes were expressed and visualized in agar gel electrophoresis using standard gene ladder. Results : A total of 41 bacterial pathogens were isolated, mainly: Bacillus spp ., Enterobacter spp., Salmonella spp ., Proteus spp ., Klebsiella spp ., E. coli, and Staphylococcus spp. The isolates were tested for their susceptibility to a panel of antibiotics using the disk diffusion method as recommended by the Clinical and Laboratory Standards Institute (CLSI). Multidrug resistance was recorded in 39 (95%) of the isolates Out of those; 5 (12.8%) were profiled genetically using multiplex Polymerase Chain Reaction assay and revealed resistome ( bla TEM in Proteus sp ., Klebsiella sp., E. coli, and Salmonella sp.) associated with beta-lactam resistance. Also, the KPC and bla OXA genes were found in Klebsiella sp. associated with carbapenem resistance. Conclusion : These findings raise a significant concern since these genes encode enzymes that provide bacteria with resistance to broad-spectrum antibiotics including carbapenem, which are the last resort in combating severe infections. Hence, it is a global health priority to control the spread of these resistant bacteria and to explore strategies for mitigating their negative impact on human health. water bacteria multidrug resistance genes Makoko community antibiotic resistance Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 INTRODUCTION Water is essential to maintain balance in human, animal, and environmental ecosystems considering the one health concept [ 1 ]. For instance, in a community like Makoko, water is the lifeblood of their daily activities: essential for their domestic use, and fishing [ 2 ]. Contrastingly, water is a significant source of public health challenges in the community [ 3 ]. Pathogenic bacteria in water pose significant public health risks, particularly in underdeveloped regions where sanitation and hygiene practices are often inadequate [ 4 , 5 ]. The World Health Organization reported that over two billion people worldwide consume water contaminated with feces, leading to diseases like cholera, typhoid, dysentery, diarrhea, and other enteric diseases [ 6 ]. Previous reports had Legionella, Mycobacteria, Vibrio cholerae , and Coliforms as the main bacterial contaminants via the faecal-oral route of water by human and animal faeces [ 7 ]. The Enterobacteriaceae family known as Coliforms include Escherichia coli ; Salmonella spp ., Enterobacter aerogenes , Shigella , and Klebsiella spp . These pathogens have been severally implicated in diseases like cholera, typhoid fever, and dysenteries in humans and livestock [ 7 ]. According to Mulamattahi et al ., [ 8 ] the prevalence of faecal Aeromonas sp. and Pseudomonas sp. might have serious health consequences for the community, particularly in individuals with low immunity. Perhaps, of greater concern is the preponderance of antimicrobial resistance occasioned by various drugs abuses of man: contamination of water bodies with antibiotics and other untreated wastes and chemical effluents. These activities spread mobile genetic elements resistant genes in bacteria, hence leading to drug resistance. Again, selection pressure coupled with interaction between pathogenic and ecological bacteria is likely to cause genetic exchange since environmental bacteria have developed resistance mechanism against antibiotics, thus making it hard to curtail, while the antibiotics continue to lose their efficacy [ 9 ]. The emergence of MDR bacteria in water bodies has generated concerns over the spread of antibiotic resistance genes and higher risk of invasive infection [ 10 ]. Furthermore, antibiotic-resistant bacteria have been identified in a wide range of waterbodies such as streams in developed and developing countries [ 11 ]. In a study by Obiakara-Amaechi et al . [ 12 ], S. aureus and Salmonella spp. were identified from the water and sediment samples collected from the Makoko and failed to screen for antibiotics resistant genes. A similar study in Pakistan reported Salmonella spp. resistance against beta-lactams and fluoroquinolones from water sources in that region. 13 Several studies in Nigeria further revealed the presence of Salmonella sp., Shigella sp., E. coli , Vibro cholera , and Pseudomonas s p. among water bodies [ 14 – 16 ]. However, only few studies have reported the prevalence of antibiotic resistant genes with bacterial pathogens from water. In Czech Republic, bla TEM, tet W, bla NDM-1, and van A resistance genes were found in the bacteria isolates obtained from wastewater [ 17 ]. Despite the palpable significance of waterbodies in the spread of resistomes many scholars reported that water ecosystem remains poorly investigated in Nigeria [ 11 , 18 ]. This study therefore aimed to reveal bacterial pathogen strains and resistomes associated with the MDR among Makoko community and the public health implications. MATERIALS AND METHODS Study design The study was a laboratory based experimental academic research. Study Area Makoko is an urban slum located in Yaba municilal area of Lagos, Nigeria. It is an impoverished community enclosed by mass of abundant Akoko trees, animals and wild swan vegetation [ 2 ]. The community is situated on a lagoon and depends on economic, domestic, and recreational activities for survival. The poor status of its inhabitants has made the community lack basic amenities such as good roads, schools, health care facilities and housing [ 2 ]. The community has high risk of waterborne diseases, as a result of the poor environmental hygiene practices and may serve as dangerous pathogen reservoir against the Lagos densely populated cities. Sample Collection Sediment and water samples were collected from three different part of the stream (Upstream, middle stream, and downstream). The water samples were purposively collected in 1liter sterile plastic bottles, while van Veen Grab sampler was used to collect the sediment sample into a Ziplock bag [ 19 ]. Sample Processing The sediment and samples were diluted serially using distilled water as the diluent and 1 mL aliquot of the selected dilutions were plated out on Muller Hinton agar (MHA) and cultured: using the standard pour plate technique as described by Terrones-Fernandez et al . [ 20 ] in duplicates and incubated at 37℃ for 24 hours. Morphologically distinct colonies of bacteria were sub-cultured onto fresh MHA to obtain pure cultures. Biochemical Characterization The isolates were speciated using biochemical tests as described by Hussain et al. [ 7 ], and identified using the guidelines of Bergey’s Manual of Determinative Bacteriology [ 21 ]. Antimicrobial Susceptibility Testing Inoculum Preparation Standard inoculum was obtained using McFarland number 5-standard to make bacterial concentration of approximately 1.5 x 10 8 CFU/ml [ 22 ]. Antibiotic susceptibility testing was carried out using recom0mended panel of antibiotics on each of the bacteria isolates applying modified Kirby-Bauer disk diffusion technique as described by CLSI [ 22 ]. Double Disk Synergy Test The MDR isolates were screened for ESBLs by placing Ceftazidime and Cefotaxime 36mm apart and a disk of Amoxicillin/clavulanate in between the two cephalosporins with a distance of 15mm from each edge on a MHA [ 23 ]. The observation of a “key-hole” zone of inhibition towards the Amoxicillin/clavulanate combination was indicative of neutralization of ESBLs enzyme Genotypic Characterization of Resistomes DNA Preparation The DNA for PCR gene amplification was carried out by introducing colonies of cell materials of overnight bacterial culture of ESBL-positive strains plated on Luria Bertani (LB) agar in 200 µl of phosphate‐buffered saline (PBS). Further procedures were applied as previously described by Obasi et al [ 24 ]. Detection of Extended Spectrum Beta-Lactamase (ESBLs) and Carbapenemase Genes For the ESBL screening, Multiplex Polymerase Chain Reaction was carried out to detect the presence of the three prominent ESBL genes (TEM, SHV and CTX-M) in the bacterial DNA samples using the following specific set of primers; TEM (Forward: 5-TTT CGT GTC GCC CTT ATT CC-3; Reverse: 5- ATC GTT GTC AGA AGT AAG TTG G-3), SHV (Forward: 5-CGC CTG TGT ATT ATC TCC CT-3; Reverse: 5- CGA GTA CTC CAC GAG ATC CT), CTX-M (Forward: 5- CGC TGT TGT TAG GAA GTG TG-3; Reverse: 5- GGC TGG GTG AAG TAA GTG AC -3). The PCR reaction was carried out using the Solis Biodyne 5X HOT FIREPol Blend Master mix. PCR was performed in a 20 µl reaction mixture, and the reaction concentration was brought down from 5x concentration to 1X concentration containing 1X Blend Master mix buffer Buffer (Solis Biodyne), 1.5 mM MgCl 2 , 200µM of each deoxynucleoside triphosphates (Solis Biodyne), 25pMol of each primer (BIOMERS, Germany), 2 unit of Hot FIREPol DNA polymerase (Solis Biodyne), Proofreading Enzyme, 2µl of the extracted DNA, and sterile distilled water was used to make up the reaction mixture. Thermal cycling was conducted in a PTC 200 gradient thermal Cycler Eppendorf for an initial denaturation of 95°C for 5 minutes followed by 35 amplification cycles of 1 minute at 95°C; 1 minute at 58°C and 1 minute at 72°C. This was followed by a final extension step of 10 minutes at 72°C. The resulting amplicons were separated on a 1.5% agarose gel and electrophoresis was carried out at 80V for 1 hour 30 minutes. After electrophoresis, DNA bands were visualized by ethidium bromide staining using a UV- Trans-illuminator. A 100bp DNA ladder was used as DNA molecular weight standard. For the screening of carbapenemase genes, the bacterial DNA were screened for three carbapenemase genes (IMP, NDM and KPC) using specific primer pairs for each gene (NDM; F: 5’-AAA ACG GCA AGA AAA AGC AG-3’, R: 5’-AAA ACG GCA AGA AAA AGC AG-3’, KPC; F: 5’-AAA ACG GCA AGA AAA AGC AG-3’, R: 3’-AAA ACG GCA AGA AAA AGC AG-3’. The reaction setup was similar to the ESBLs gene detection but in this case, only IMP and NDM were multiplexed due to similarity in their annealing temperature while KPC was ran as a uniplex. Thermal cycling condition was carried out at an initial denaturation of 95°C for 5 minutes followed by 35 amplification cycles of 1 minute at 95°C; 1 minute at 52°C and 1 minute at 72°C. This was followed by a final extension step of 10 minutes at 72°C. The resulting amplicons were separated on a 2% agarose gel and electrophoresis was carried out at 80V for 1 hour 30 minutes. After electrophoresis, DNA bands were visualized by ethidium bromide staining using a UV- Trans-illuminator. A 100bp DNA ladder was used as DNA molecular weight standard [ 24 ]. RESULTS A total number of 93 bacteria were isolated; however, 41 pathogenic strains were selected for further studies. Figure 1 presents the prevalence of pathogenic isolates studied. Figures 2 and 3 present the antibiogram of the gram-negative and gram-positive isolates respectively. All of the isolates were resistant to more than three classes of antibiotics except for the two Bacillus species that were resistant to just two classes of antibiotics. Table 1 , presents the antibacterial resistance index shown by the isolates and ranged from 0.2(the least) to 1(highest); being the reciprocal of the number of the antibiotics that were resistant to a particular strain of bacteria. Table 1 The resistance index range Isolates (No) No of Antibiotics No Resistant Range Index Range E. coli (3) 12 6–7 0.5–0.6 Enterobacter spp.(6) 12 5–8 0.4–0.7 Klebsiella spp. (4) 12 6–7 0.5–0.8 Acinetobacter spp. 12 12 1 Shigella spp. 12 4 0.3 Salmonella spp. (4) 12 3–8 0.3–0.7 Proteus spp. (4) 12 4–12 0.3-1 Pseudomonas spp. 12 8 0.7 Serratia spp. (2) 12 6–9 0.5–0.8 Bacillus spp. (13) 11 2–7 0.2–0.6 S. aureus (2) 11 3–7 0.3–0.6 ( The MAR index was calculated and interpreted according to Sazon and Migo (2020), and is the reciprocal of the number of antibiotics to which an isolate was resistant and the total number of antibiotics tested ). Table 2 presents the genotype of ESBLs expressed phenotypically among the organisms studied, and mainly showed TEM resistome. Isolate Code Organism PCR Code Gene Expressed CTX TEM SHV UW25 Proteus sp. 1 - + - UW157 Salmonella sp. 2 - - - MW172 Klebsiella sp. 3 - + - DS163 Shigella sp. 4 - + - UW158 Escherichia coli 5 - + - Legend : - = Negative; + = Positive Table 3 shows the genotype of the Carbapenemase producing bacteria studied Code Isolates PCR No Carbapenem Occurrence KPC IMP NDM OXA UW25 Proteus sp. 1 - - - - UW157 Salmonella sp. 2 - - - - MW172 Klebsiella sp. 3 + - - + DS163 Shigella sp. 4 - - - - UW158 Escherichia coli 5 - - - - Legend : Negative = - ; Positive = + The result showed that Klebsiella sp. was positive to bla KPC and bla OXA. The gene expression on the agar gel electrophoresis are shown on Fig. 5 . Discussion From the observations in the course of our study, Makoko community is an improvised area that lacks basic amenities. Hence, the release of waste materials, faeces, and unfettered solid wastes directly into the community stream (Oko-Agbon stream), this observation is in line with the previous assessment done by Simon et al. [ 2 ] and Obiakara-Amaechi et al. [ 12 ]. Therefore, due to the direct release of sewage and other pollutants, the number of pathogenic bacteria in the stream may most likely surge leading to contaminations and spread of resistomes among bacteria Concomitantly, due to poor quality of the stream water, the community possesses high risk of waterborne disease and low efficient of antibiotics due to increase of multi-drug resistant bacteria [ 25 ]. Also, the growing populations are a significant factor in the rising environmental health issues which escalate poor hygiene practices including dumping wastes directly into the stream, as reported by Ayeni [ 14 ]. Of the 41 bacteria purposively selected for further analysis, the most common isolates were gram-negative bacilli (n = 26), followed by gram positive bacilli (n = 13), and gram-positive cocci (n = 2) which correlated with the study by OV et al . [ 26 ], wherein the identified bacteria isolates were predominantly gram-positive cocci, Gram-negative bacilli, and Gram-positive bacilli respectively. Contrastingly, in this study, Bacilli organisms were identified to be more predominant in the water sample. This report agrees with similar research carried out by Obioma et al. [ 27 ]. Their report had it that Bacillus species are known to be ubiquitous, and that they produce spores that are resistant to environmental strain. Bacteria with such flexibility are known to thrive in almost all the environment, including water bodies [ 27 ]. This study revealed the presence of S. aureus which was similar to the findings of Obiakara-Amaechi et al. [ 12 ] from water and sediment in Makoko. The study suggested that the inherent pathogenicity of S. aureus is intricately linked to a set of virulence factors and its propensity to develop resistance against diverse classes of antibiotics and the same was also reported by Adekanmbi et al. [ 28 ]. The presence of E. coli, Klebsiella, Enterobacter, Salmonella, Shigella and Vibrio species are particularly worrisome, These are well documented pathogens that not only renders the water unsuitable for human consumption, but also gives rise to significant health apprehensions. Other scholars expressed such concerns [ 29 , 30 ]. These findings attribute the contamination to the indiscriminate disposal of organic waste and the pollution of the water source through fecal contamination. Furthermore, the presence of these bacteria in the stream signifies a serious indication of possible disease outbreaks and potential epidemics. Furthermore, noteworthy pathogens like Pseudomonas spp. and S. aureus have been associated with skin and soft tissue infections such as wound infections, as reported by Bessa et al . [ 31 ]. There is therefore a grave public health concern since the water studied is the only water source for domestic and commercial fishing business of the community. In this study, most of the bacteria showed resistance to 3 or more classes of antibiotics. This could be as the result of spread of resisitomes and mobile genetic elements as suggested by Tersagh et al. [ 32 ]. There exist reports of direct discharge of effluents to the stream and pharmaceutical industrial waste, including night soil men activities [ 32 , 11 ]. These reports raise the likelihood that Makoko is a vulnerable community. The MAR index proves invaluable for gauging the prevalence of resistant bacteria in a given environment. In this report (table 6), the MAR indices ranged from 0.2–1. Those with a MAR index exceeding 0.4 typically stemmed from human fecal sources, while those below 0.4 may have originated from non-human fecal contamination [ 33 ]. The study found that 92% of the bacterial isolates had a MAR value > 0.2, suggesting potential exposure to multiple antibiotics. A similar study by Poonia et al . [ 33 ] showed a lesser (36.2%) percentage of bacteria showed > 0.2 MAR index. This high resistance might be a result of selective pressure from antibiotic use in treating infections in animals or humans living in the slum. Furthermore, high resistance of Escherichia coli to more than three antibiotics was observed. Five bacteria with significant health implications were found to be MDR and harboured ESBLs and Carbapenemase genes. Of the 5, E. coli, Proteus sp., Shigella sp., and Klebsiella sp. were positive for bla TEM genes. Adesoji and Ogunjobi [ 34 ] reported the presence of the bla TEM gene in E. coli, Proteus sp., and Klebsiella sp. from water sources in western Nigeria and Lagos is in this region. This could imply high abuse of the third-generation cephalosporins. Furthermore, Klebsiella spp. was also positive for bla OXA and bla KPC, similarly, Ahmed et al [ 35 ] and Suzuki et al. [ 36 ] respectively reported the bla KPC gene from water samples. These reports indicate the broader distribution of the bla genes. Adesoji and Ogunjobi [ 34 ] reported that bacterial strains are capable of inducing life-threatening illnesses and could trigger widespread outbreaks. For example, Habib [ 37 ] documented that the presence of CTX-M and TEM genes in Klebsiella spp . and E. coli have been linked to nosocomial infections and outbreaks of diarrhea. Although this study did not find E. coli with the bla CTX gene, Pandey et al. [ 38 ] and Suzuki et al. [ 36 ] reported bla CTX among E. coli which mainly trail feca-oral route; several studies have reported similar findings [ 36 , 39 ]. Conclusion This study reveals an alarming 92% MDR bacterial strain from the study site. This finding highlights the urgent need for improved water quality management and infection control measures to allay the spread of MDR bacteria. Also, due to the increased rate of susceptibility to Gentamycin and Levofloxacin, antibiotics can be recommended for effective treatment of diseases caused by pathogenic bacterial isolates. Also, access to clean water and proper sanitation can significantly reduce the prevalence and spread of bacterial pathogens and implementation of policies that can regulate the discharge of untreated sewage and wastes directly into the stream; thus, reducing the environment reservoirs of MDR bacteria. The need for continued surveillance and in-depth investigation into the genetic determinants of antibiotic resistance in this community’s bacterial population is underscored. Declarations Acknowledgments The authors are grateful for the following people for their technical support, Dr. Apeh Isaac Ojima (University of Nigeria Nnsukka), Ms Elizabeth Abidemi Adebayor (Trinity University Yaba), Mr Peter and Dr. Fowora (Nigerian Institute of Medical Research). Authors’ contributions This work was carried out through the collaborative efforts of all the authors. Authors MEA, AJO, and AMC initiated the idea. Author AJO designed the study and wrote the protocol. Authors FOT, ECA, and MCA revised the protocol and study design. Authors MEA and AJO were involved in the sample collection and laboratory analysis. ECA, FOT, and SJO supervised the laboratory procedures. AJO wrote the draft for the manuscript. ECA, FOT, and SJO reviewed the manuscript. All authors read and approved the final manuscript. Funding The study was self-funded. Data availability All data generated in the course of the study are included in this article. Ethical approval The study was approved by ethical and faculty officers of Basic Medical and Applied Sciences of Trinity University Yaba as an academic program. Consent for publication Not applicable Consent to participate declaration Not applicable Competing interests No conflict of interest declared Author details 1 Department of Biological Sciences, Trinity University, Yaba, Lagos State, Nigeria. 2 Microbiology Department, Nigerian Institute of Medical Research, Yaba, Lagos State, Nigeria. References Olowe BM, Oluyege AO, Famurewa O. Prevalence of waterborne diseases and microbial assessment of drinking water quality in Ado-Ekiti and its environs, southwestern, Nigeria. Br Microbiol Res J. 2016;12(2):1–13. https://doi.org/10.9734/bmrj/2016/22444 . Simon R, Adegoke AK, Adewale BA. 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Contamination of water resources by pathogenic bacteria. AMB Express. 2014;4(1). https://doi.org/10.1186/s13568-014-0051-x . Akinduti PA, Olasehinde GI, Ejilude O, Taiwo OS, Obafemi YD. Fecal carriage and Phylo-Diversity of Community-Acquired bla TEM Enteric Bacilli in Southwest Nigeria. Infect Drug Resist. 2018;11:2425–33. https://doi.org/10.2147/IDR.S178243 . Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 03 Feb, 2026 Read the published version in BMC Microbiology → Version 1 posted Editorial decision: Revision requested 09 Sep, 2025 Reviews received at journal 13 Aug, 2025 Reviewers agreed at journal 05 Aug, 2025 Reviewers agreed at journal 05 Aug, 2025 Reviews received at journal 01 Aug, 2025 Reviewers agreed at journal 12 Jul, 2025 Reviewers agreed at journal 23 May, 2025 Reviewers agreed at journal 29 Apr, 2025 Reviewers invited by journal 24 Apr, 2025 Editor assigned by journal 18 Apr, 2025 Editor invited by journal 15 Apr, 2025 Submission checks completed at journal 15 Apr, 2025 First submitted to journal 15 Apr, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6341991","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":447329584,"identity":"200b60f4-d9cc-4236-a137-03750bd1dd5f","order_by":0,"name":"John O. Apeh","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA8ElEQVRIiWNgGAWjYHACNgYGHgYGAxDxAcRlJ0UL4wwQl5koLQwQLcw8IBYhLfyzm489+CBjI2/O3nvwsc2vbfJ8zAyMHz7m4NYicedYuuEMnjTDnT3nko1z+24btjEzMEvO3IbHmhs5ZtI8PIcTDECM3J7bjEAtbMy8eLTI38j/BtFy/435b8ue2/YEtQANZ4PawmPGzPDjdiJBLYZ3jplJgvyy4UxesmRvw+3kNmbGZrx+kbvd/EziY4+NvMHxswc//Phz23Z+e/PBDx/xeV8CiBl7oBzGNjDZgEc9VAvDDxjvD37Fo2AUjIJRMDIBAGN7T1wyTMOZAAAAAElFTkSuQmCC","orcid":"","institution":"Trinity University Sabo-Yaba","correspondingAuthor":true,"prefix":"","firstName":"John","middleName":"O.","lastName":"Apeh","suffix":""},{"id":447329586,"identity":"3883d6c3-ce38-4690-9c17-ce432c85d52a","order_by":1,"name":"Emmanuel A. Meshach","email":"","orcid":"","institution":"Trinity University Sabo-Yaba","correspondingAuthor":false,"prefix":"","firstName":"Emmanuel","middleName":"A.","lastName":"Meshach","suffix":""},{"id":447329588,"identity":"2e4460bf-0bae-41f8-b690-4bb87f1cff76","order_by":2,"name":"Oluwatosin T. Fagbemi","email":"","orcid":"","institution":"Trinity University Sabo-Yaba","correspondingAuthor":false,"prefix":"","firstName":"Oluwatosin","middleName":"T.","lastName":"Fagbemi","suffix":""},{"id":447329590,"identity":"9ec8440a-3b0a-4ead-b238-a557e8509ee7","order_by":3,"name":"Modupeade C. Adetunji","email":"","orcid":"","institution":"Trinity University Sabo-Yaba","correspondingAuthor":false,"prefix":"","firstName":"Modupeade","middleName":"C.","lastName":"Adetunji","suffix":""},{"id":447329591,"identity":"d5a28724-6eae-4a52-a90a-ca55801f5247","order_by":4,"name":"Michael N. Aransiola","email":"","orcid":"","institution":"Trinity University Sabo-Yaba","correspondingAuthor":false,"prefix":"","firstName":"Michael","middleName":"N.","lastName":"Aransiola","suffix":""},{"id":447329592,"identity":"5785a230-7fd1-479b-afa8-b2a9d90e3a94","order_by":5,"name":"Joseph O. Shaibu","email":"","orcid":"","institution":"Nigerian Institute of Medical Research","correspondingAuthor":false,"prefix":"","firstName":"Joseph","middleName":"O.","lastName":"Shaibu","suffix":""},{"id":447329593,"identity":"4030b65a-f92f-4abf-b44e-7c952aa929d4","order_by":6,"name":"Christian A. Enwuru","email":"","orcid":"","institution":"Nigerian Institute of Medical Research","correspondingAuthor":false,"prefix":"","firstName":"Christian","middleName":"A.","lastName":"Enwuru","suffix":""}],"badges":[],"createdAt":"2025-03-31 06:09:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6341991/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6341991/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12866-026-04793-3","type":"published","date":"2026-02-03T15:58:49+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":81542041,"identity":"4780f00a-f50e-4b51-98ea-d48bd2622e92","added_by":"auto","created_at":"2025-04-28 11:17:21","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":105665,"visible":true,"origin":"","legend":"\u003cp\u003eFrequency of the identified Gram negative and positive bacterial isolates\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6341991/v1/bfc54b16b11280282fa5e29d.png"},{"id":81544780,"identity":"fbc81fd9-6fd6-4747-87e2-c283aad23f71","added_by":"auto","created_at":"2025-04-28 11:33:22","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":346855,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSusceptibility and Resistance percentage for Gram-negative bacterial isolates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLegend\u003c/strong\u003e: NF=Nitrofurantoin; CXM=Cefuroxime; CRO= Ceftriaxone sulbactam; ACX= Ampiclox; ZEM= Cefexime; LBC= Levofloxacin; AUG= Amoxicillin clavulanic acid; CTX= Cefotaxime; IMP= Imipenem; OFX= Ofloxacin; GN= Gentamycin; NA= Nalidixic acid.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6341991/v1/62d126b7866fcdbe3fa3aeff.png"},{"id":81543722,"identity":"bee1d030-b98f-4742-b8ec-d96ea00af57b","added_by":"auto","created_at":"2025-04-28 11:25:22","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":235813,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSusceptibility and Resistance percentage for Gram-positive bacterial isolates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLegend:\u003c/strong\u003e CXM=Cefuroxime; CRO= Ceftriaxone sulbactam; AZN= Azithromycin; ERY= Erythromycin; ZEM= Cefexime; LBC= Levofloxacin; AUG= Amoxicillin clavulanic acid; CTX= Cefotaxime; IMP= Imipenem; OFX= Ofloxacin; GN= Gentamycin\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6341991/v1/d2b0d6a344025d6d782ee444.png"},{"id":81544779,"identity":"801749b6-0731-4544-8a29-06f80536c579","added_by":"auto","created_at":"2025-04-28 11:33:22","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":343761,"visible":true,"origin":"","legend":"\u003cp\u003eTEM (409bp) gene expressed at Lanes 1, 3, 4 and 5 after PCR on 1%Agarose gel electrophoresis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKey\u003c/strong\u003e: Lane M: 100-bp DNA ladder control and Lane N: Negative control\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6341991/v1/c2cca63dde6f9ca2d4fa6a55.png"},{"id":81542057,"identity":"d4fd806a-75eb-4ddc-8cea-dc77075a2cda","added_by":"auto","created_at":"2025-04-28 11:17:22","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":597357,"visible":true,"origin":"","legend":"\u003cp\u003eOXA and KPC genes after PCR on 1%Agarose gel electrophoresis. Lane M: 100-bp DNA ladder; Lane 3 is positive OXA gene at (585bp) and KPC gene at (340bp); Lane N: Negative control\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6341991/v1/bba06976b2017f01ffdd9f98.png"},{"id":102234355,"identity":"cc7fa64e-5059-4d3f-8aa8-235f5a19979b","added_by":"auto","created_at":"2026-02-09 16:10:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2498832,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6341991/v1/99c3ca37-1ac8-4ed9-8849-6cf5573ad30f.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Resistomes Associated with Transmissible Bacterial Pathogens in Water and Sediments from Oko-Agbon Stream, Makoko, Lagos, Nigeria","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eWater is essential to maintain balance in human, animal, and environmental ecosystems considering the one health concept [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. For instance, in a community like Makoko, water is the lifeblood of their daily activities: essential for their domestic use, and fishing [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Contrastingly, water is a significant source of public health challenges in the community [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Pathogenic bacteria in water pose significant public health risks, particularly in underdeveloped regions where sanitation and hygiene practices are often inadequate [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The World Health Organization reported that over two billion people worldwide consume water contaminated with feces, leading to diseases like cholera, typhoid, dysentery, diarrhea, and other enteric diseases [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePrevious reports had Legionella, Mycobacteria, \u003cem\u003eVibrio cholerae\u003c/em\u003e, and Coliforms as the main bacterial contaminants via the faecal-oral route of water by human and animal faeces [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. The Enterobacteriaceae family known as Coliforms include \u003cem\u003eEscherichia coli\u003c/em\u003e; \u003cem\u003eSalmonella spp\u003c/em\u003e., \u003cem\u003eEnterobacter aerogenes\u003c/em\u003e, \u003cem\u003eShigella\u003c/em\u003e, and \u003cem\u003eKlebsiella spp\u003c/em\u003e. These pathogens have been severally implicated in diseases like cholera, typhoid fever, and dysenteries in humans and livestock [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. According to Mulamattahi \u003cem\u003eet al\u003c/em\u003e., [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e] the prevalence of faecal \u003cem\u003eAeromonas\u003c/em\u003e sp. and \u003cem\u003ePseudomonas\u003c/em\u003e sp. might have serious health consequences for the community, particularly in individuals with low immunity.\u003c/p\u003e \u003cp\u003ePerhaps, of greater concern is the preponderance of antimicrobial resistance occasioned by various drugs abuses of man: contamination of water bodies with antibiotics and other untreated wastes and chemical effluents. These activities spread mobile genetic elements resistant genes in bacteria, hence leading to drug resistance. Again, selection pressure coupled with interaction between pathogenic and ecological bacteria is likely to cause genetic exchange since environmental bacteria have developed resistance mechanism against antibiotics, thus making it hard to curtail, while the antibiotics continue to lose their efficacy [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe emergence of MDR bacteria in water bodies has generated concerns over the spread of antibiotic resistance genes and higher risk of invasive infection [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Furthermore, antibiotic-resistant bacteria have been identified in a wide range of waterbodies such as streams in developed and developing countries [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. In a study by Obiakara-Amaechi \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eSalmonella\u003c/em\u003e spp. were identified from the water and sediment samples collected from the Makoko and failed to screen for antibiotics resistant genes. A similar study in Pakistan reported \u003cem\u003eSalmonella\u003c/em\u003e spp. resistance against beta-lactams and fluoroquinolones from water sources in that region.\u003csup\u003e13\u003c/sup\u003e Several studies in Nigeria further revealed the presence of \u003cem\u003eSalmonella\u003c/em\u003e sp., \u003cem\u003eShigella\u003c/em\u003e sp., \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eVibro cholera\u003c/em\u003e, and \u003cem\u003ePseudomonas s\u003c/em\u003ep. among water bodies [\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. However, only few studies have reported the prevalence of antibiotic resistant genes with bacterial pathogens from water. In Czech Republic, \u003cem\u003ebla\u003c/em\u003eTEM, \u003cem\u003etet\u003c/em\u003eW, \u003cem\u003ebla\u003c/em\u003eNDM-1, and \u003cem\u003evan\u003c/em\u003eA resistance genes were found in the bacteria isolates obtained from wastewater [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDespite the palpable significance of waterbodies in the spread of resistomes many scholars reported that water ecosystem remains poorly investigated in Nigeria [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. This study therefore aimed to reveal bacterial pathogen strains and resistomes associated with the MDR among Makoko community and the public health implications.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy design\u003c/h2\u003e \u003cp\u003eThe study was a laboratory based experimental academic research.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eStudy Area\u003c/h3\u003e\n\u003cp\u003eMakoko is an urban slum located in Yaba municilal area of Lagos, Nigeria. It is an impoverished community enclosed by mass of abundant Akoko trees, animals and wild swan vegetation [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The community is situated on a lagoon and depends on economic, domestic, and recreational activities for survival. The poor status of its inhabitants has made the community lack basic amenities such as good roads, schools, health care facilities and housing [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The community has high risk of waterborne diseases, as a result of the poor environmental hygiene practices and may serve as dangerous pathogen reservoir against the Lagos densely populated cities.\u003c/p\u003e\n\u003ch3\u003eSample Collection\u003c/h3\u003e\n\u003cp\u003eSediment and water samples were collected from three different part of the stream (Upstream, middle stream, and downstream). The water samples were purposively collected in 1liter sterile plastic bottles, while van Veen Grab sampler was used to collect the sediment sample into a Ziplock bag [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eSample Processing\u003c/h3\u003e\n\u003cp\u003eThe sediment and samples were diluted serially using distilled water as the diluent and 1 mL aliquot of the selected dilutions were plated out on Muller Hinton agar (MHA) and cultured: using the standard pour plate technique as described by Terrones-Fernandez \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] in duplicates and incubated at 37℃ for 24 hours. Morphologically distinct colonies of bacteria were sub-cultured onto fresh MHA to obtain pure cultures.\u003c/p\u003e\n\u003ch3\u003eBiochemical Characterization\u003c/h3\u003e\n\u003cp\u003eThe isolates were speciated using biochemical tests as described by Hussain \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], and identified using the guidelines of Bergey\u0026rsquo;s Manual of Determinative Bacteriology [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eAntimicrobial Susceptibility Testing\u003c/h2\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003eInoculum Preparation\u003c/h2\u003e \u003cp\u003eStandard inoculum was obtained using McFarland number 5-standard to make bacterial concentration of approximately 1.5 x 10\u003csup\u003e8\u003c/sup\u003eCFU/ml [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAntibiotic susceptibility testing was carried out using recom0mended panel of antibiotics on each of the bacteria isolates applying modified Kirby-Bauer disk diffusion technique as described by CLSI [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003eDouble Disk Synergy Test\u003c/h3\u003e\n\u003cp\u003eThe MDR isolates were screened for ESBLs by placing Ceftazidime and Cefotaxime 36mm apart and a disk of Amoxicillin/clavulanate in between the two cephalosporins with a distance of 15mm from each edge on a MHA [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. The observation of a \u0026ldquo;key-hole\u0026rdquo; zone of inhibition towards the Amoxicillin/clavulanate combination was indicative of neutralization of ESBLs enzyme\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eGenotypic Characterization of Resistomes\u003c/h2\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003eDNA Preparation\u003c/h2\u003e \u003cp\u003eThe DNA for PCR gene amplification was carried out by introducing colonies of cell materials of overnight bacterial culture of ESBL-positive strains plated on Luria Bertani (LB) agar in 200 \u0026micro;l of phosphate‐buffered saline (PBS). Further procedures were applied as previously described by Obasi \u003cem\u003eet al\u003c/em\u003e [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eDetection of Extended Spectrum Beta-Lactamase (ESBLs) and Carbapenemase Genes\u003c/h2\u003e \u003cp\u003eFor the ESBL screening, Multiplex Polymerase Chain Reaction was carried out to detect the presence of the three prominent ESBL genes (TEM, SHV and CTX-M) in the bacterial DNA samples using the following specific set of primers; TEM (Forward: 5-TTT CGT GTC GCC CTT ATT CC-3; Reverse: 5- ATC GTT GTC AGA AGT AAG TTG G-3), SHV (Forward: 5-CGC CTG TGT ATT ATC TCC CT-3; Reverse: 5- CGA GTA CTC CAC GAG ATC CT), CTX-M (Forward: 5- CGC TGT TGT TAG GAA GTG TG-3; Reverse: 5- GGC TGG GTG AAG TAA GTG AC -3). The PCR reaction was carried out using the Solis Biodyne 5X HOT FIREPol Blend Master mix. PCR was performed in a 20 \u0026micro;l reaction mixture, and the reaction concentration was brought down from 5x concentration to 1X concentration containing 1X Blend Master mix buffer Buffer (Solis Biodyne), 1.5 mM MgCl\u003csub\u003e2\u003c/sub\u003e, 200\u0026micro;M of each deoxynucleoside triphosphates (Solis Biodyne), 25pMol of each primer (BIOMERS, Germany), 2 unit of Hot FIREPol DNA polymerase (Solis Biodyne), Proofreading Enzyme, 2\u0026micro;l of the extracted DNA, and sterile distilled water was used to make up the reaction mixture. Thermal cycling was conducted in a PTC 200 gradient thermal Cycler Eppendorf for an initial denaturation of 95\u0026deg;C for 5 minutes followed by 35 amplification cycles of 1 minute at 95\u0026deg;C; 1 minute at 58\u0026deg;C and 1 minute at 72\u0026deg;C. This was followed by a final extension step of 10 minutes at 72\u0026deg;C. The resulting amplicons were separated on a 1.5% agarose gel and electrophoresis was carried out at 80V for 1 hour 30 minutes. After electrophoresis, DNA bands were visualized by ethidium bromide staining using a UV- Trans-illuminator. A 100bp DNA ladder was used as DNA molecular weight standard. For the screening of carbapenemase genes, the bacterial DNA were screened for three carbapenemase genes (IMP, NDM and KPC) using specific primer pairs for each gene (NDM; F: 5\u0026rsquo;-AAA ACG GCA AGA AAA AGC AG-3\u0026rsquo;, R: 5\u0026rsquo;-AAA ACG GCA AGA AAA AGC AG-3\u0026rsquo;, KPC; F: 5\u0026rsquo;-AAA ACG GCA AGA AAA AGC AG-3\u0026rsquo;, R: 3\u0026rsquo;-AAA ACG GCA AGA AAA AGC AG-3\u0026rsquo;. The reaction setup was similar to the ESBLs gene detection but in this case, only IMP and NDM were multiplexed due to similarity in their annealing temperature while KPC was ran as a uniplex. Thermal cycling condition was carried out at an initial denaturation of 95\u0026deg;C for 5 minutes followed by 35 amplification cycles of 1 minute at 95\u0026deg;C; 1 minute at 52\u0026deg;C and 1 minute at 72\u0026deg;C. This was followed by a final extension step of 10 minutes at 72\u0026deg;C. The resulting amplicons were separated on a 2% agarose gel and electrophoresis was carried out at 80V for 1 hour 30 minutes. After electrophoresis, DNA bands were visualized by ethidium bromide staining using a UV- Trans-illuminator. A 100bp DNA ladder was used as DNA molecular weight standard [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cp\u003eA total number of 93 bacteria were isolated; however, 41 pathogenic strains were selected for further studies. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e presents the prevalence of pathogenic isolates studied.\u003c/p\u003e\n\u003cp\u003eFigures \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e present the antibiogram of the gram-negative and gram-positive isolates respectively. All of the isolates were resistant to more than three classes of antibiotics except for the two Bacillus \u003cem\u003especies\u003c/em\u003e that were resistant to just two classes of antibiotics.\u003c/p\u003e\n\u003cp\u003eTable\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, presents the antibacterial resistance index shown by the isolates and ranged from 0.2(the least) to 1(highest); being the reciprocal of the number of the antibiotics that were resistant to a particular strain of bacteria.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eThe resistance index range\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIsolates (No)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth colspan=\"2\" align=\"left\"\u003e\n \u003cp\u003eNo of Antibiotics No Resistant Range\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIndex Range\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e(3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u0026ndash;7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.5\u0026ndash;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eEnterobacter\u003c/em\u003e spp.(6)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u0026ndash;8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.4\u0026ndash;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eKlebsiella\u003c/em\u003e spp. (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u0026ndash;7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.5\u0026ndash;0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAcinetobacter\u003c/em\u003e spp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eShigella\u003c/em\u003e spp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eSalmonella\u003c/em\u003e spp. (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u0026ndash;8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3\u0026ndash;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eProteus\u003c/em\u003e spp. (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u0026ndash;12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3-1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003ePseudomonas\u003c/em\u003e spp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eSerratia\u003c/em\u003e spp. (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u0026ndash;9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.5\u0026ndash;0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus\u003c/em\u003e spp. (13)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u0026ndash;7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2\u0026ndash;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eS. aureus\u003c/em\u003e (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u0026ndash;7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3\u0026ndash;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e(\u003c/strong\u003e \u003cstrong\u003eThe MAR index was calculated and interpreted according to Sazon and Migo (2020), and is the reciprocal of the number of antibiotics to which an isolate was resistant and the total number of antibiotics tested\u003c/strong\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n \u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003epresents the genotype of ESBLs expressed phenotypically among the organisms studied, and mainly showed TEM resistome.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIsolate Code\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eOrganism\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePCR Code\u003c/p\u003e\n \u003c/th\u003e\n \u003cth colspan=\"3\" align=\"left\"\u003e\n \u003cp\u003eGene Expressed\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eCTX\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTEM\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSHV\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUW25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eProteus\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUW157\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eSalmonella\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMW172\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eKlebsiella\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDS163\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eShigella\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUW158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eEscherichia coli\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003eLegend\u003c/strong\u003e: - = Negative; + = Positive\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eshows the genotype of the Carbapenemase producing bacteria studied\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCode\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIsolates\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePCR No\u003c/p\u003e\n \u003c/th\u003e\n \u003cth colspan=\"3\" align=\"left\"\u003e\n \u003cp\u003eCarbapenem Occurrence\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eKPC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eIMP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNDM\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eOXA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUW25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eProteus\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUW157\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eSalmonella\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMW172\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eKlebsiella\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDS163\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eShigella\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUW158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eEscherichia coli\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003e\u003cstrong\u003eLegend\u003c/strong\u003e: Negative = \u003cstrong\u003e-\u003c/strong\u003e ; Positive = \u003cstrong\u003e+\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe result showed that \u003cem\u003eKlebsiella\u003c/em\u003e sp. was positive to \u003cem\u003ebla\u003c/em\u003eKPC and \u003cem\u003ebla\u003c/em\u003eOXA. The gene expression on the agar gel electrophoresis are shown on Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eFrom the observations in the course of our study, Makoko community is an improvised area that lacks basic amenities. Hence, the release of waste materials, faeces, and unfettered solid wastes directly into the community stream (Oko-Agbon stream), this observation is in line with the previous assessment done by Simon \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e] and Obiakara-Amaechi \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Therefore, due to the direct release of sewage and other pollutants, the number of pathogenic bacteria in the stream may most likely surge leading to contaminations and spread of resistomes among bacteria\u003c/p\u003e \u003cp\u003eConcomitantly, due to poor quality of the stream water, the community possesses high risk of waterborne disease and low efficient of antibiotics due to increase of multi-drug resistant bacteria [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Also, the growing populations are a significant factor in the rising environmental health issues which escalate poor hygiene practices including dumping wastes directly into the stream, as reported by Ayeni [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOf the 41 bacteria purposively selected for further analysis, the most common isolates were gram-negative bacilli (n\u0026thinsp;=\u0026thinsp;26), followed by gram positive bacilli (n\u0026thinsp;=\u0026thinsp;13), and gram-positive cocci (n\u0026thinsp;=\u0026thinsp;2) which correlated with the study by OV \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e], wherein the identified bacteria isolates were predominantly gram-positive cocci, Gram-negative bacilli, and Gram-positive bacilli respectively. Contrastingly, in this study, Bacilli organisms were identified to be more predominant in the water sample. This report agrees with similar research carried out by Obioma \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Their report had it that Bacillus species are known to be ubiquitous, and that they produce spores that are resistant to environmental strain. Bacteria with such flexibility are known to thrive in almost all the environment, including water bodies [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThis study revealed the presence of \u003cem\u003eS. aureus\u003c/em\u003e which was similar to the findings of Obiakara-Amaechi \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e] from water and sediment in Makoko. The study suggested that the inherent pathogenicity of \u003cem\u003eS. aureus\u003c/em\u003e is intricately linked to a set of virulence factors and its propensity to develop resistance against diverse classes of antibiotics and the same was also reported by Adekanmbi \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe presence of \u003cem\u003eE. coli, Klebsiella, Enterobacter, Salmonella, Shigella and Vibrio\u003c/em\u003e species are particularly worrisome, These are well documented pathogens that not only renders the water unsuitable for human consumption, but also gives rise to significant health apprehensions. Other scholars expressed such concerns [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. These findings attribute the contamination to the indiscriminate disposal of organic waste and the pollution of the water source through fecal contamination. Furthermore, the presence of these bacteria in the stream signifies a serious indication of possible disease outbreaks and potential epidemics. Furthermore, noteworthy pathogens like \u003cem\u003ePseudomonas\u003c/em\u003e spp. and \u003cem\u003eS. aureus\u003c/em\u003e have been associated with skin and soft tissue infections such as wound infections, as reported by Bessa \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. There is therefore a grave public health concern since the water studied is the only water source for domestic and commercial fishing business of the community.\u003c/p\u003e \u003cp\u003eIn this study, most of the bacteria showed resistance to 3 or more classes of antibiotics. This could be as the result of spread of resisitomes and mobile genetic elements as suggested by Tersagh \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. There exist reports of direct discharge of effluents to the stream and pharmaceutical industrial waste, including night soil men activities [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. These reports raise the likelihood that Makoko is a vulnerable community.\u003c/p\u003e \u003cp\u003eThe MAR index proves invaluable for gauging the prevalence of resistant bacteria in a given environment. In this report (table 6), the MAR indices ranged from 0.2\u0026ndash;1. Those with a MAR index exceeding 0.4 typically stemmed from human fecal sources, while those below 0.4 may have originated from non-human fecal contamination [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. The study found that 92% of the bacterial isolates had a MAR value\u0026thinsp;\u0026gt;\u0026thinsp;0.2, suggesting potential exposure to multiple antibiotics. A similar study by Poonia \u003cem\u003eet al\u003c/em\u003e. [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e] showed a lesser (36.2%) percentage of bacteria showed\u0026thinsp;\u0026gt;\u0026thinsp;0.2 MAR index. This high resistance might be a result of selective pressure from antibiotic use in treating infections in animals or humans living in the slum. Furthermore, high resistance of \u003cem\u003eEscherichia coli\u003c/em\u003e to more than three antibiotics was observed.\u003c/p\u003e \u003cp\u003eFive bacteria with significant health implications were found to be MDR and harboured ESBLs and Carbapenemase genes. Of the 5, \u003cem\u003eE. coli, Proteus\u003c/em\u003e sp., \u003cem\u003eShigella\u003c/em\u003e sp., and \u003cem\u003eKlebsiella\u003c/em\u003e sp. were positive for \u003cem\u003ebla\u003c/em\u003eTEM genes. Adesoji and Ogunjobi [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] reported the presence of the \u003cem\u003ebla\u003c/em\u003eTEM gene in \u003cem\u003eE. coli, Proteus\u003c/em\u003e sp., and \u003cem\u003eKlebsiella\u003c/em\u003e sp. from water sources in western Nigeria and Lagos is in this region. This could imply high abuse of the third-generation cephalosporins. Furthermore, \u003cem\u003eKlebsiella spp.\u003c/em\u003e was also positive for \u003cem\u003ebla\u003c/em\u003eOXA and \u003cem\u003ebla\u003c/em\u003eKPC, similarly, Ahmed \u003cem\u003eet al\u003c/em\u003e [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e] and Suzuki \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] respectively reported the \u003cem\u003ebla\u003c/em\u003eKPC gene from water samples. These reports indicate the broader distribution of the \u003cem\u003ebla\u003c/em\u003e genes. Adesoji and Ogunjobi [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] reported that bacterial strains are capable of inducing life-threatening illnesses and could trigger widespread outbreaks. For example, Habib [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] documented that the presence of CTX-M and TEM genes in \u003cem\u003eKlebsiella spp\u003c/em\u003e. and \u003cem\u003eE. coli\u003c/em\u003e have been linked to nosocomial infections and outbreaks of diarrhea. Although this study did not find \u003cem\u003eE. coli\u003c/em\u003e with the \u003cem\u003ebla\u003c/em\u003eCTX gene, Pandey \u003cem\u003eet al.\u003c/em\u003e[\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e] and Suzuki \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] reported \u003cem\u003ebla\u003c/em\u003eCTX among \u003cem\u003eE. coli\u003c/em\u003e which mainly trail feca-oral route; several studies have reported similar findings [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study reveals an alarming 92% MDR bacterial strain from the study site. This finding highlights the urgent need for improved water quality management and infection control measures to allay the spread of MDR bacteria. Also, due to the increased rate of susceptibility to Gentamycin and Levofloxacin, antibiotics can be recommended for effective treatment of diseases caused by pathogenic bacterial isolates. Also, access to clean water and proper sanitation can significantly reduce the prevalence and spread of bacterial pathogens and implementation of policies that can regulate the discharge of untreated sewage and wastes directly into the stream; thus, reducing the environment reservoirs of MDR bacteria. The need for continued surveillance and in-depth investigation into the genetic determinants of antibiotic resistance in this community\u0026rsquo;s bacterial population is underscored.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful for the following people for their technical support, Dr. Apeh Isaac Ojima (University of Nigeria Nnsukka), Ms Elizabeth Abidemi Adebayor (Trinity University Yaba), \u0026nbsp;Mr Peter and Dr. Fowora (Nigerian Institute of Medical Research).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was carried out through the collaborative efforts of all the authors. Authors MEA, AJO, and AMC initiated the idea. Author AJO designed the study and wrote the protocol. Authors FOT, ECA, and MCA revised the protocol and study design. Authors MEA and AJO were involved in the sample collection and laboratory analysis. ECA, FOT, and SJO supervised the laboratory procedures. AJO wrote the draft for the manuscript. ECA, FOT, and SJO reviewed the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was self-funded.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated in the course of the study are included in this article.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was approved by ethical and faculty officers of Basic Medical and Applied Sciences of Trinity University Yaba as an academic program.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate declaration\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo conflict of interest declared\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor details\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003eDepartment of Biological Sciences, Trinity University, Yaba, Lagos State, Nigeria.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003eMicrobiology Department, Nigerian Institute of Medical Research, Yaba, Lagos State, Nigeria.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eOlowe BM, Oluyege AO, Famurewa O. 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Contamination of water resources by pathogenic bacteria. AMB Express. 2014;4(1). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s13568-014-0051-x\u003c/span\u003e\u003cspan address=\"10.1186/s13568-014-0051-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkinduti PA, Olasehinde GI, Ejilude O, Taiwo OS, Obafemi YD. Fecal carriage and Phylo-Diversity of Community-Acquired bla TEM Enteric Bacilli in Southwest Nigeria. Infect Drug Resist. 2018;11:2425\u0026ndash;33. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2147/IDR.S178243\u003c/span\u003e\u003cspan address=\"10.2147/IDR.S178243\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"water, bacteria, multidrug resistance genes, Makoko community, antibiotic resistance","lastPublishedDoi":"10.21203/rs.3.rs-6341991/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6341991/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eWater is essential for life; however, it serves as the route of transmission for many infectious pathogens. The increasing prevalence of multidrug-resistant (MDR) bacteria in water samples poses a significant public health risk.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAim\u003c/strong\u003e: This study aimed to characterize and analyze the molecular profile of selected MDR bacteria isolated from water and sediment samples from the Oko-Agbon stream in the Makoko community.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethod\u003c/strong\u003e: The bacteria were identified using conventional methods and subsequently subjected to antimicrobial susceptibility testing using Kirby-Bauer disk diffusion technique. The MDR strains were screened for ESBLs and Carbapenemase genes phenotypically by double disk synergy method and genotypically by PCR amplification. The specific genes were expressed and visualized in agar gel electrophoresis using standard gene ladder.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: A total of 41 bacterial pathogens were isolated, mainly: \u003cem\u003eBacillus \u003c/em\u003espp\u003cem\u003e., Enterobacter \u003c/em\u003espp.,\u003cem\u003e Salmonella \u003c/em\u003espp\u003cem\u003e., Proteus \u003c/em\u003espp\u003cem\u003e., Klebsiella \u003c/em\u003espp\u003cem\u003e.,\u003c/em\u003e \u003cem\u003eE. coli, \u003c/em\u003eand \u003cem\u003eStaphylococcus \u003c/em\u003espp. The isolates were tested for their susceptibility to a panel of antibiotics using the disk diffusion method as recommended by the Clinical and Laboratory Standards Institute (CLSI). Multidrug resistance was recorded in 39 (95%) of the isolates Out of those; 5 (12.8%) were profiled genetically using multiplex Polymerase Chain Reaction assay and revealed resistome (\u003cem\u003ebla\u003c/em\u003eTEM in \u003cem\u003eProteus \u003c/em\u003esp\u003cem\u003e., Klebsiella \u003c/em\u003esp.,\u003cem\u003e E. coli, and Salmonella\u003c/em\u003e sp.) associated with beta-lactam resistance. Also, the KPC and \u003cem\u003ebla\u003c/em\u003eOXA genes were found in \u003cem\u003eKlebsiella \u003c/em\u003esp. associated with carbapenem resistance.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: These findings raise a significant concern since these genes encode enzymes that provide bacteria with resistance to broad-spectrum antibiotics including carbapenem, which are the last resort in combating severe infections. Hence, it is a global health priority to control the spread of these resistant bacteria and to explore strategies for mitigating their negative impact on human health.\u003c/p\u003e","manuscriptTitle":"Resistomes Associated with Transmissible Bacterial Pathogens in Water and Sediments from Oko-Agbon Stream, Makoko, Lagos, Nigeria","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-28 11:17:17","doi":"10.21203/rs.3.rs-6341991/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-09T23:48:44+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-13T13:28:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"114221972308274758524393678357016425422","date":"2025-08-05T16:20:52+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"227175927245328179278613413683843013187","date":"2025-08-05T16:18:33+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-02T00:03:52+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"10267708742127447261630927015265852985","date":"2025-07-12T23:16:25+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"41498467639514943440201804413888946744","date":"2025-05-23T06:10:50+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"67019835226142617263524546311975107066","date":"2025-04-29T12:16:28+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-04-24T08:04:29+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-04-18T11:52:09+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-04-15T20:09:58+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-04-15T14:29:09+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Microbiology","date":"2025-04-15T14:27:58+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"82064171-fbba-4924-8fe8-2168f22934d3","owner":[],"postedDate":"April 28th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-02-09T16:05:07+00:00","versionOfRecord":{"articleIdentity":"rs-6341991","link":"https://doi.org/10.1186/s12866-026-04793-3","journal":{"identity":"bmc-microbiology","isVorOnly":false,"title":"BMC Microbiology"},"publishedOn":"2026-02-03 15:58:49","publishedOnDateReadable":"February 3rd, 2026"},"versionCreatedAt":"2025-04-28 11:17:17","video":"","vorDoi":"10.1186/s12866-026-04793-3","vorDoiUrl":"https://doi.org/10.1186/s12866-026-04793-3","workflowStages":[]},"version":"v1","identity":"rs-6341991","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6341991","identity":"rs-6341991","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}