Prevalence, Molecular Characterization, Pathogenicity and Antimicrobial Susceptibility Pattern of Clostridioides Difficile From Clinical Samples in South Eastern Nigeria.

Prevalence, Molecular Characterization, Pathogenicity and Antimicrobial Susceptibility Pattern of Clostridioides Difficile From Clinical Samples in South Eastern Nigeria. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Prevalence, Molecular Characterization, Pathogenicity and Antimicrobial Susceptibility Pattern of Clostridioides Difficile From Clinical Samples in South Eastern Nigeria. Eddison Igho Oghonyon, Malachy C. Ugwu, Charles O. Esimone, Anthony Ik Onah This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4955956/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The prevalence, molecular characterization, pathogenicity and antibioticsensitivity pattern of Clostridioides difficile from clinical samples in South-Eastern Nigeria wereevaluated in this study. Stool samples (450) were collected from patientsinsome selected public and private-owned clinics in the five South-Eastern states. Standard bacteriological methods were employed for enumeration of total anaerobes. Culture of Clostridioides difficile isolates were performed on Clostridium difficile differential agar (supplemented with C. difficile selective supplement). Biochemical tests like catalase, oxidase and reverse CAMP test were conducted before extraction of genomic bacterial DNA for suspected C. difficile isolates. Purification and amplification of bacterial DNA was carried out on 2% agarose gel. Amplified bacteria DNA was sequenced and blasted on the National Centre for Biotechnology Information (NCBI) website. Antibiotics susceptibility was carried out for C. difficile isolates using the Kirby-Bauer disc diffusion technique. PCR technique was employed for the detection of virulence ( tcdA, tcdB, cdtA and cdtB ) and resistance genes ( tetS, tetA and ermB ) in C. difficile isolates. Anaerobe counts for stool samples obtained showed that samples had a mean count of 5.63±0.09 log 10 CFU/g (Anambra state) while counts of 5.61±0.11 log 10 CFU/g and 5.65±0.07 log 10 CFU/g were obtained from Ebonyi and Enugu States. The antibacterial sensitivity revealed that the isolates were sensitive to gentamicin, meropenem and amoxicillin-clavulanic acid. The isolates were resistant to tetracyclines and erythromycin, and the multiple antibiotic resistance index of theisolates showed that the multi-drug resistant isolates had a MAR index of 0.44. All (14 C. difficile isolates) (100%) of C. difficile isolates were found to possess tcdB genes, and 69.56% expressed tcdA genes. One (4.34%) of the C. difficile isolatespossessed cdtA and cdtB genes for binary toxin production. prevalence molecular characterization pathogenicity Antibiogram Clostridioides difficile South-East INTRODUCTION Standard healthcare facilities are essential for survival in a world dominated by superbugs,which have also been implicated in infections and the re-occurrence of infection even after the treatment regimen. Pathogenic organisms have recorded unparalleled success in their ability to cause diseases as well as induce therapeutic failure through drug resistance (Adegokeet al., 2016 ). Clostridioides difficile has been reported as the leading cause of healthcare-associated diarrhea (Markantonis et al., 2024 ). Antibiotic exposure, which reduces the population of non-pathogenic anaerobes that normally inhabit the gut, has been pointed out asthe most important risk factor for Clostridioides difficile infection (CDI) (Markantonis et al., 2024 ).Thus, CDI occurs when there is a shift in the microbial flora of the colon, allowingtoxin-producing strains of the Gram-positive, spore-forming, anaerobic bacillus toproliferate (Smits et al., 2016; Markantonis et al., 2024 ). Other events implicated in gut microbiota disruption includeproton pump inhibitors (PPI) use and inflammatory bowel diseases (Buddle and Fagan, 2023 ; Xiaolu et al., 2024 ). Researchers have attributed the pathogenicity of C. difficile to the two major virulence factors produced by toxigenic C. difficile strains, enterotoxin A ( TcdA ) and cytotoxin B ( TcdB ) (Buddle and Fagan 2023 ; Xiaolu et al., 2024 ) On September 16th, 2013, the Centre for Disease Control (CDC) declared Clostridioides difficile as a threat with “urgent” concern to public health in the United States due to the identification of antimicrobial resistance among human and animal-derived Clostridioides difficile isolates to drugs commonly used in humans and also to their increased rates of associated patient mortality since the early 2000s(CDC, 2013). In all the studies, some strains of C. difficile appear to be species-specific, while other strains were isolated from multiple species, including man. The organism was initially described as part of the normal flora in stool samples from healthy infants in 1935 (Parija, 2023 ), as well as its detection in significant numbers in healthy asymptomatic children (Khoder et al., 2019 ). It was afterwardidentified as a pathogen associated with several diseases, and even now, in hospitals and long-term care facilities, it has become the most common cause of diarrhea, causing the loss of billions of dollars in healthcarecosts (Panhwar, 2022).The symptoms of infection usually appear mild in some cases, and in others, they appear to be life-threatening or fatal(Mondal, 2019 ). Diarrhea associated with C. difficile is accompanied by the passage of occult blood or mucus in the stool, but hematochezia and melena are other rare symptoms (Sheen et al., 2021 ; Xiaoluet al., 2024 ). Certain other symptoms are found in less than 50% of infected patients, including abdominal discomfort, fever, and peripheral leukocytosis (Voutsinaset al., 2020 ).Disease caused by C. difficile is linked most commonly to nosocomial infections in humans, especially after empiric antibiotic treatment. Humans develop a spectrum of diseases when infected with some strains of C. difficile . It has been shown that not all people who are on prolonged antibiotic medication, as well as those above the age of 60 years, are susceptible to CDI(Jaceket.al, 2019 ). The advent of some virulent new strains of C. difficile has triggered an increase in CDI rates even in younger populations originally thought to be at low risk of contracting C. difficile infection (Doyle, 2013 ).To date, little is known about CDI in Southeastern Nigeria as inadequate attention has been given to the understanding of the molecular epidemiology of CDI in Nigeria, a high-burden country.Thus, the research was aimedatdeterminingthe prevalence, molecular characterization, pathogenicity, and antimicrobial susceptibility pattern of Clostridioides difficile from clinical samples in the South-Eastern states of Nigeria. METHOD Study Area The study was conducted after obtaining due ethical approval from the ethical committee of the ChukwuemekaOdumegwuOjukwu University Teaching Hospital (COUTH), Amaku, Awka,for the collection of stool samples in the hospital in Awka andneighbouring states. Sample Collection The clinical samples were collected from selectedpublic hospitals.Stool samples were collected in red cap universal bottles from COOUTH in Anambra, Federal Teaching Hospital Abakaliki in Ebonyi, Abia State University Teaching Hospital in Abia, Imo State University Teaching Hospital in Imo and Enugu State Teaching Hospital in Enugu. Samples were collected from patients for analysis in the clinical laboratory, after which they werecollected and preserved in an ice pack for further analysis. The samples were collected and someof the challeges in collection includes: untimely collection, patient unwillingness to cooperate, cross contamination risk,health and safety issues A total of 450 stool samples were collected and evaluated in this study. The sample size was determined using the sample size determination formula below: \(\:Samplesize\:\left(N\right)=\:{(Z}^{2\:}P\left(1-P\right))÷{D}^{2}\) ………….Eq. 1 where: N = Sample size, Z = z score (1.969), P = prevalence (0.45) andD = 0.05 Isolation and Identification of Clostridium difficile For the isolation of Clostridioides difficile , all samples were pre-treated (heat shocked and alcohol shocked) to reduce competing flora and allow the growth of spore-forming bacilli. AStandard microbiological procedure was employed using Clostridium difficile differential agar CM 0601 (Oxoid) supplemented with Clostridium difficile selective supplement SR0096 (Oxoid) (Bridson, 2006 ). Stock solutions were prepared for the samples by weighing 20g of the stool samples into 180 ml of sterile saline water in a conical flask. A ten-fold serial dilution from the stock solution was made after mixing. From the fifth tube, 2 ml of the suspension was added to a well-labelled tube containing 2 ml of absolute ethanol (alcohol/ethanol shock method). The mixture was rocked gently to allow for homogenization, and it was left to stand for an hour before plating (of 100µl) was carried out. Heat treatment, also known as heat shocking, was employed by heating the tubes containing a mixture of samples to boil for 5 minutes. The tubes were then allowed to cool before plating was carried out on the media supplemented with Clostridium difficile supplement (Oxoid).The formula employed for the dilution factor is given below in Eq. (2) $$\:Dilution\:factor\:=\:\frac{final\:volume}{aliquot\:volume\:}\left(2\right)$$ $$\:where:final\:volume=aliquot\:volume\:\left(sample\:volume\right)\:+diluent\:volume$$ Enumeration of the bacterial/fungal isolates was carried out using the formula delineated by Willey et al. ( 2008 ), and it is shown in Eq. (3) below. $$\:\frac{cfu}{g}=\frac{number\:of\:colonies\:x\:dilution\:factor}{volume\:of\:inoculum\:}\:\:\:\:\:\:\:\:\:\:\:\left(3\right)\:$$ The cultural, morphological, microscopic and biochemical properties of the isolates were evaluated after Gram and spore staining was done to select the positive isolates for both stains. Biochemical tests like catalase, oxidase, indole, urease and sugar fermentation tests were carried out to identify Clostridium species. The identity of Clostridioides difficile was finalized following blasting of sequences obtained using 16sRNA universal primer sets. Antibiotic Susceptibility Test The identified C. difficile were subjected to standard antibacterial susceptibility testing (AST). The antibiotic discs (Oxoid UK) containing the following antibiotics Meropenem (10 µg), Erythromycin (15 µg), Metronidazole (50 µg), Amoxicillin/clavulanic acid (20/10 µg), Clindamycin (20 µg), Gentamicin (10 µg), Ciprofloxacin (5 µg), Vancomycin (30 µg), and Tetracycline (30 µg) was used. Briefly, an18-24 hours bacterial culture wascultured on MHA. The inoculum corresponding to 1.5 x 10 8 cells/ml McFarland standard wasstreaked using a sterile loop onto the MHA plates before the antibiotic discs were aseptically added with extreme care to the plates with the aid of sterile forceps. The susceptibility results were recorded after incubation for 24 hours at 37 ºC and interpreted usingClinical Laboratory Standards Institute breakpoints (CLSI, 2020). Multiple Antibiotic Resistance Index determination The MAR index was determined by the formula by Chitanandet al. ( 2010 ): \(\:MARindex=\frac{y}{nx}\) ………..Eq. 4 $$\:wherey=numberofresistancescored,\:n=numberofisolates$$ $$\:x=total\:number\:of\:antibioitics$$ Extraction of DNA DNA was extracted using boiling method described by Chakravorty et al . (2007). 2 ml of 24 h. broth culture of C. difficile was transferred to tube and centrifuged at high speed for 5 min. The supernatant was discarded after which 200 µL of sterile distilled water (SDW) was added to the pellets and vortexed using a vortex mixer for 1 min before it was heated at 100 o C for 15 mins. Lastly, it was thereafter centrifuge at high speed for 2 minutes. The supernatant from the second spin was now regarded as the pure DNA and from which, 10 µL of DNA was used for amplification of the gene by PCR. Table 1 Primer sets for virulence genes Genes Primer Sequence (5’-3’) Conc. (µM) Product size (bp) TcdA tcdA-F tcdA-R GTATGGATAGGTGGAGAAGTCAGTG CGGTCTAGTCCAATAGAGCTAGGTC 0.025 632 tcdB tcdB-F tcdB-R GAAGATTTAGGAAATGAAGAAGGTGA AACCACTATATTCAACTGCTTGTCC 0.01 441 cdtA cdtA-F cdtA-R ATGCACAAGACTTACAAAGCTATAGTG CGAGAATTTGCTTCTATTTGATAATC 0.2 260 cdtB cdtB-F cdtB-R ATTGGCAATAATCTATCTCCTGGA CCAAAATTTCCACTTACTTGTGTTG 0.5 179 The presence of genes which code for virulence ( tcdA, tcdB, cdtA and cdtB ) was determined using standardized primer sets in the table above. Table 2 Primer sets for resistance genes Genes Primer Sequence (5’-3’) Conc. (µM) Product size (bp) erm(B ErmB-F ErmB-R GAAAAGGTACTCAACCAAATA AGTAACGGTACTTAAATTGTTTAC 0.1 639 tet(S) TetS-F TetS-R ATCAAGATATTAAGGAC TTCTCTATGTGGTAATC 0.1 573 tet(A) tet(A)- F tet(A)- R TTGGCATTCTGCATTCACTC GTATAGCTTGCCGGAAGTCG 0.1 494 The genes coding for erythromycin ( erm B) and tetracycline resistance ( tet A and tetS ) were screened using polymerase chain reaction (PCR). The previously extracted bacterial DNA sample was used for the analysis. Agarose gel electrophoresis was carried out using each primer (forward and reverse primers) for the target genes (Call et al., 2003 ; Florez et al ., 2014). A thermocycler was used to determine an optimal annealing temperature for the specific binding of the primer set to the DNA template. RESULTS Table 3 Total anaerobic bacterial count of stool samples obtained from patients in different states Location Number of Samples Anaerobe count (Log 10 CFU/g) p-value (s) Anambra 90 5.63 ± 0.09 a 0.168 Ebonyi 90 5.62 ± 0.11 ab 0.168 Enugu 90 5.65 ± 0.07 a 0.168 Imo 90 5.37 ± 0.19 b 0.066 Abia 90 5.45 ± 0.09 b 0.066 The results of the anaerobic stool sample count from Eastern Nigeria areshown in Table 3 above.The total anaerobe count (log 10 CFU/g) for stool samples obtained from patients in different states showed that stool samples obtained from patients in Anambra had counts of 5.63 ± 0.09 log 10 CFU/g. In contrast, samples from Ebonyi, Enugu and Imo states had counts of 5.61 ± 0.11 log 10 CFU/g, 5.65 ± 0.07 log 10 CFU/g and 5.36 ± 0.19 log 10 CFU/g, respectively. Plate 1. Agarose gel electrophoresis of bacterial DNA isolated from stool samples Table 4 Identity of bacterial isolates (anaerobes) from stool samples from Eastern Nigeria Sample Code Source Identity Query Cover(%) Homology (%) Accession number anStool12 Stool Clostridioides difficile 100 100.00 MH888201.1 ebstool05 Stool Clostridioides difficile 98 91.4 CP019858.1 Imstool15 Stool Clostridioides difficile 99 99.8 CP019860.1 anStool21 Stool Clostridioides difficile 99 96.75 CP025047.1 anStool22 Stool Clostridioides difficile 97 90.35 CP020424.2 abstool12 Stool Clostridioides difficile 100 100.00 MH888202.1 abstool17 Stool Clostridioides difficile 99 99.8 CP019860.1 imstool83 Stool Clostridioides difficile 91 94.6 CP028361.1 imstool49 Stool Clostridioides difficile 91 94.6 CP028361.1 ebstool23 Stool Clostridioides difficile 91 94.6 CP028361.1 ebstool33 Stool Clostridioides difficile 99 99.8 CP019860.1 ebstool1 Stool Clostridioides difficile 99 96.75 CP025047.1 abstool12 Stool Clostridioides difficile 97 90.35 CP020424.2 anstool70 Stool Clostridioides difficile 100 100.00 MH888202.1 The identity and Phylogenetic relatedness of anaerobic bacterial isolates obtained from stool samples in Eastern Nigeria are shown in Table 4 above. Table 5 Prevalence of anaerobes from stool samples using standard cultural methods Samples Samples Number Anambra Ebonyi Enugu Imo Abia Total Sample(study) Study Total (%) Stool Samples 90 55(61) 45(50) 50(55) 41(46) 50(55) 450 241(54) The prevalence of anaerobes from stool samples using standard cultural methods (Table 5 ) revealed that all stool samples but Imo (46%) had anaerobic counts equal to or greater than 50% following a cultural phenotypic standard. In comparison, a total of 54% was obtained in the entire study. Table 6 Prevalence of Clostridioides difficile from stool samples in the study Samples Sample Number Anambra Ebonyi Enugu Imo Abia Total sample (study) Study Total (%) Stool Samples 90 4(44.4) 2(22.2) 2(22.22) 3(33.3) 3(33.3) 450 14(3.11) The prevalence of Clostridioides difficile from stool samples (Table 6 ) revealed a total study prevalence of 3.11% for stool samples. Stool samples from Ebonyi and Enugu states had a prevalence of 2.22% apiece. In comparison, Imo and Abia states had a 3.33% prevalence of C. difficile from stool samples, with Anambra state having the highest prevalence of 4.44%. Table 7 Percentage Recovery of C. difficile from stool samples using cultural methods Sample Number Heat Treatment (%) Alcohol Treatment (%) Study Total (%) Stool Samples 450 5(1.11) 14(3.11) 14(3.11) The prevalence of C. difficile recovered by the heat treatment method (1.11%) was found to be lower than the percentage recovered via alcohol treatment (3.11%). Table 8 a: Antibiotic sensitivity profile to common antibiotics Code Samples MEM ERY MET AMC CL CN CIP VA TET anStool12 Stool S R S R S S S R R ebStool05 Stool S S S R S S S R R imStool15 Stool S R R S R S R S R anStool21 Stool S S S S S S S S R anStool22 Stool S R S S S S R S R abStool12 Stool R S R S R S S S R abStool77 Stool S R R S S S R S R imStool83 Stool S R S S R S R S S imstool49 Stool S R R S R S R S R ebstool23 Stool R R R S S S S S R enstool33 Stool S R S S S S S S R enstool1 Stool S R S S R S R S R abStool14 Stool S S S R S S S R S anstool70 Stool S S S S S S R S S Keys: Meropenem (10 µg), Erythromycin (15 µg), Metronidazole (50 µg), Amoxicillin/clavulanic acid (20/10 µg), Clindamycin (20 µg), Gentamicin (10 µg), Ciprofloxacin (5 µg), Vancomycin (30 µg), Tetracycline (30 µg) Antibacterial sensitivity pattern and percentage of resistance/susceptibility of C. difficile isolates stool samples to common antibiotics showed susceptibility to carbapenems (82.61%), aminoglycosides (86.98%) and Β-Lactam/combination agents (73.91%). Notable resistance was observed for tetracycline (73.91%), macrolides (73.91%) and fluoroquinolones (43.48%), respectively. Table 8 b: Percentage of antibiotic resistance and susceptibility profile of common antibiotics Antibiotics Classes Antibiotics Susceptibility profile n = 09 Sensitive (%) Resistant (%) Carbapenems MEM 19(82.61) 3(17.39) Macrolides ERY 6(26.09) 17(73.91) Nitroimidazole MET 16(69.56) 7(30.43) Β-Lactam/combination agents AMC 17(73.91) 6(26.09) Lincosamide CL 15(65.22) 8(34.78) Aminoglycosides CN 20(86.98) 3(13.02) Fluoroquinolone CIP 13(56.52) 10(43.48) Glycopeptides VA 16(69.56) 7(30.43) Tetracyclines TET 6(26.09) 17(73.91) Key: Meropenem (10 µg), Erythromycin (15 µg), Metronidazole (50 µg), Amoxicillin/clavulanic acid (20/10 µg), Clindamycin (20 µg), Gentamicin (10 µg), Ciprofloxacin (5 µg), Vancomycin (30 µg), Tetracycline (30 µg). Table 9 Multiple antibiotic resistance distribution of C. difficile isolates from clinical sources Isolate Code Antibacterial Classes Number of Antibiotics Resistance Phenotypes Resistant phenotypes (%) MAR Index anBE7, ebstool05 9 9 AMCR, VAR, TETR 2(8.69) 0.33 enLV4, ebstool23 9 9 MEMR, ERYR, METR, TETR 2(8.69) 0.44 ebLV5 9 9 ERYR, CNR, TETR 1(4.35) 0.33 anStool12 9 9 ERYR, AMCR, VAR, TETR 1(4.35) 0.44 imstool15, imstool49 9 9 ERYR, METR, CLR, CIPR, TETR, 2(8.69) 0.55 anstool21 9 9 TETR 1(4.35) 0.11 anstool222 9 9 ERYR, CIPR, TETR 1(4.35) 0.33 abstool12 9 9 MEMR, ERYR, METR, CLR, TETR 1(4.35) 0.55 enstool33 9 9 ERYR, TETR 1(4.35) 0.22 enstool1 9 9 ERYR, CLR, CIPR, TETR 1(4.35) 0.44 abstool14 9 9 AMCR, VAR 1(4.35) 0.22 anstool70 9 9 CIPR 1(4.35) 0.11 Keys : Meropenem (10 µg), Erythromycin (15 µg), Metronidazole (50 µg), Amoxicillin/clavulanic acid (20/10 µg), Clindamycin (20 µg), Gentamicin (10 µg), Ciprofloxacin (5 µg), Vancomycin (30 µg), Tetracycline (30 µg) The multiple antibiotic resistance phenotypic distribution of C. difficile isolates from clinical samples (Table 9 ) showed that isolates with a MAR index of 0.44 had the highest resistance phenotypes. In contrast, isolates with a MAR index of 0.33 had the next highest resistance phenotypes. Table 10 Prevalence of virulence genes in isolates of C. difficile obtained from stool samples Lanes Code Samples tcdA tcdB cdtA cdtB Total (%) 1 anStool12 Stool + + - - 2(50) 2 ebstool05 Stool + + - - 2(50) 3 Imstool15 Stool + + - - 2(50) 4 anstool21 Stool - + - - 1(25) 5 anstool22 Stool + + + + 4(100) 6 abstool12 Stool + + - - 2(50) 7 abstool77 Stool + + - - 2(50) 8 imstool83 Stool - + - - 1(25) 9 imstool49 Stool + + - - 2(50) 10 ebstool23 Stool - + - - 2(50) 11 enstool33 Stool + + - - 2(50) 12 enstool11 Stool - + - - 1(25) 13 abstool12 Stool + + - - 2(50) 14 anstool70 Stool + + - - 2(50) Total Study(%) 16(69.56) 23(100.00) 1(4.34) 1(4.34) All (100%) of C. difficile isolates were found to possess tcdB genes. About tcdA genes, 10 of 14 positive isolates were found to possess the virulence genes, making a total of 69.56% of isolates with positive results for the presence of the genes. None of the isolated C. difficile strains but one possessed cdtA and cdtB genes, thus giving a percentage of 4.34% apiece for the aforementioned genes. The prevalence of virulence genes in Table 10 revealed that 3 isolates possessed 25% (one of the four) of virulence genes analyzed in the study, while 10 isolates possessed two of the four virulence genes (50%) Table 11 Prevalence of resistant genes to erythromycin and tetracycline found in isolates of C. difficile obtained from stool samples Lanes Code Samples TetA tetS ErmB Total(%) 1 anStool12 Stool + + + 3(100) 2 ebstool05 Stool + + - 2(67) 3 Imstool15 Stool + + + 3(100) 4 anstool21 Stool + + - 2(67) 5 anstool22 Stool + + + 3(100) 6 abstool12 Stool + + + 3(100) 7 abstool77 Stool + + + 3(100) 8 imstool83 Stool - - + 1(33) 9 imstool49 Stool + + + 3(100) 10 ebstool23 Stool + + + 3(100) 11 enstool33 Stool + + + 3(100) 12 enstool1 Stool + + + 3(100) 13 abstool12 Stool - - - 0(0) 14 anstool70 Stool - - - 0(0) Total study (%) 17(73.91) 17(73.91) 17(73.91) . Agarose gel electrophoresis of resistance genes present in C. difficile obtained from stool samples is shown in Table 11 . In contrast, erm(B) gene PCR products amplified from Clostridioides difficile isolates were found in 73.91%; it was also the same percentage that was present for tetracycline resistance ( tet(S) and tet(A) genes). Nine isolates were found to possess all resistance genes analyzed in the study . DISCUSSION The majority of bacteria (anaerobes) in the stool are commensals, which oftendo not harm humans unless there is an underlying risk factor or the case of a compromised immune system. The bacterium of interest in this study ( C. difficile ) is regarded as both a commensal organism and a pathogen to domestic animals (Doyle, 2013). Most of these anaerobes in the gut that have been isolated from the feces are not competitive, and their population can only explode, or they can thrive better in the gut after an array of antibiotic treatments which could destroy much of the intestinal normal flora (Doyle, 2013). The statistical comparison of the total anaerobic count in stool samples showedno significant difference ( p>0.05 ) between samples obtained from Ebonyi and other Eastern states in the study. However, there was a significant difference between samples obtained from Anambra and those from Imo and Abia states, respectively. The bacterial identity, query cover, and percentage homology were written against the confirmed blasted identity of the isolates,including accession numbers obtained from the National Centre for Biotechnology Information (NCBI) website. The 3.11% prevalence of Clostridiumdifficile obtained in this study is less than several reported prevalences in the literature. This implies that the probability of infection due to C. difficile would be very low.However, the presence of the pathogen (even in its low prevalence) in this study is an indication that there is a likely chance of a CDI outbreak if proper measures are not in place to curtail the infection. Following reports by Warriner et al . (2017), C. difficile was previously thought to be clinically associated alone. Still, with advanced scientific methods, it is now being muted as a community-acquired infection. The prolonged survival of spores of C. difficile in the environment might also increase the possibility of animal and food contamination which is a potential source of infection(Weese, 2010). In Europe, a higher prevalence had also been reported thanthe one obtained in this study, howbeit for hospitalized patients with even higher reports of toxigenic strains of C. difficile recovered from patients’stool (Martirosian et al ., 1993; Shehabi et al., 2001). Nonetheless, findings in this study corroborated the report of Mori and Aoki (2015) and Deane et al. (2021),who opined that carriage rates of C. difficile in healthy adults varied from 0% to 3% in Europe to up to 15% in Japan. Following cultural phenotypic methods, the percentage recovery of C. difficile from stool samples revealed that while both treatment methods (heat and alcohol treatments) yielded positive C. difficile growth, the alcohol shock method was found to be better at recovering the bacterium isolates from clinical samples. The findings in this study about how the spores of the bacterium were recovered from both treatment processes agreed with the findings of Lawley et al . (2009). They reported that spores are difficult to eradicate due to their ability to resist radiation, heat and chemicals (such as alcohols), which are usually employed as disinfectants in both hospital and community settings. In this study, alcohol shock samples (3.11%) had a better recovery rate than heat shock samples (1.11%).However, the susceptibility to Carbapenems and aminoglycosides by most isolates of C. difficile in this study was at variance with what was observed in other studies (Rodriguez-Palacios et al ., 2007; Simango and Mwakurudz, 2008; Jöbstl et al ., 2010) which separately reported some levels of resistance to the antibiotics by the bacterium of interest.It is noteworthy that some of these antibiotics, as mentioned above, are commonly used for the treatment of diarrhea in humans. In vitro, resistance in C. difficile to several classes of antimicrobial agents hasbeen recognized and reported by Gerding (2004). Thus, there is the possibility of a high risk for CDI to occur in healthcare settings and the community at large. This finding corroborated the report of Freeman et al. (2009), who asserted that the resistance of C. difficile to several antibacterial agents may not be essential for infection because the organismisrelatively susceptible to high-risk antimicrobial agents. It has been reported that the major pathogenic mechanism of C. difficile is the production of cytotoxin B and enterotoxin A, which are respectively encoded by tcdB and tcdA genes, co-located in a 19.6 kb region of the chromosome named pathogenicity loci (Paloc) along with other regulatory genes (Freeman et al ., 2009;Buddle and Fagan 2023; Xiaolu et al., 2024).Toxin A causes diarrhea, and Toxin B is cytotoxic to the colonic cells. Virtually all studies where C. difficile have been successfully recovered have screened for the ability of the isolates to have had at least one of the three toxins or genes ( tcdA, tcdB, cdtA/B ) needed to fulfill the criteria for virulence/toxigenicity (Rodriguez-Palacios et al ., 2014). The findings in this study (69.56% and 100.00% for tcdA and tcdB genes respectively) are in agreement with the report of Shokoohizadeh et al . (2021), who opined that the genes for enterotoxin production tcdA are found in approximately 70% of C. difficile isolates, while the genes for cytotoxin production are found in all strains of C. difficile isolates. The severity of the disease caused by the bacterium is conferred by the presence of the genes ( cdtA and cdtB genes), which code for the production of binary toxins (Rupnik, 2009; Bacci et al ., 2011). The pathogenicity of C. difficile is mainly due to the presence of two large protein toxins (toxin A and toxin B) and the fully described binary toxin (Chandrasekaran and Db, 2017). Toxin A is a potent enterotoxinthat causes the accumulation of fluid in the gut, is cytotoxic to cells in tissue culture, and isperhaps lethal to experimental animals (Kuehne et al., 2010).Toxin B has been reported to be a 1,000-fold more potentcytotoxin than toxin A (but is not an enterotoxin) in original animal studies. Still,there are reports that some strains produce toxin B and not toxin A,which causes severe diarrhea in humans (Barbut et al ., 2007). More so, the reports of C. difficile strains are toxin A–negative and toxin B–positive, whichhave been associated with human diseases described in certain epidemics around the world (Johnson et al ., 2001). The variation in the findings in this study could be a function of geographical differences and environmental conditions with respect to cultural methods and methods used for the analysis of the bacterium. The findings in this study are consistent with the reports from literature, which have it that the bacterium ( C. difficile ) from humans or animals are commonly resistant to macrolides (erythromycin) and tetracyclines as well as fluoroquinolones (moxifloxacin) and the lincosamides (clindamycin).The findings in this study are consistent with the reports from theliterature, which have it that the bacterium ( C. difficile ) from humans or animals are commonly resistant to macrolides (erythromycin) and tetracyclines as well as fluoroquinolones (moxifloxacin) and the lincosamides (clindamycin) (Gerding, 2004; Simango and Mwakurudz, 2008; Jöbstl et al ., 2010; ). Also consistent with the finding in this study was the report by Knetsch et al . (2018), who opined that the global C. difficile population contained a broad array of antibiotic-resistance genes encoding resistance to tetracycline and erythromycin. This study provides baseline data for the prevalence of Clostridium difficile in clinical samples in Southeast Nigeria. Chances of severe infections for this pathogen in the studied location will be rare owing to the low prevalence and non-possession of genes for binary toxin production in most of the isolates in the study. The study has re-enacted the idea that the alcohol shock method for isolation of Clostridium difficile is more effective than the heat-shock method. Pathogenic Clostridium difficile isolates in the studied location were multi-drug resistant. CONCLUSION This study evaluated the prevalence, molecular characterization, pathogenicity, and antibiotic sensitivity patterns of Clostridium difficile in South-Eastern Nigeria, revealing high sensitivity to gentamicin, meropenem, and amoxicillin-clavulanic acid, but resistance to tetracyclines and erythromycin, with significant expression of tcdB and tcdA virulence genes. Declarations CONFLICT OF INTEREST The authors declare that they have no conflict of interest. FUNDING No funding received for this study. Ethics Approval and Consent to Participate This research was carriedout in line with the principles of the Declaration of Helsinki. Ethical approval for this research was obtained from Ethical Committee, Chuckwuemeka Odumegwu Ojukwu University Teachicng Hosptial Amaku, Awka, Anambra State, Nigeria. The ethics approval number is COOUTH/CMAC/ETH.C/VOL.1/0035 and the approval was granted on 1/08/2018 . The committee reviewed and approved all aspects of the study, including the study design, recruitment methods, and data collection procedures, ensuring that they were in compliance with ethical standards and guidelines. The consent to Participate was obtained from all individuals that participated in this research. Individuals were given detailed information regarding the research’s objectives, procedures, potential risks, and benefits. They were informed that participation was voluntary, that they could withdraw from the research at any given time without any punishment, and their data would be under confidentiality. All these were done prior to sample colections from the individuals Consent for Publication This was done by informing all participants involved in this research about the nature of the data/material to be published, the purpose of its publication, and the potential accessibility of the publication to the general public. The individuals agrees that their details may be used in the manuscript titled “ Prevalence, Pathogenicity and Antimicrobial Susceptibility Pattern of Clostridioides Difficile From Clinical Samples in South Eastern Nigeria ” and published in BMC Microbiology Journal They gave their full consent knowing fully well that this redearch will contribute greatly to scientific knowledge, which could be made available in print and online formats. .Individuals were given full assurance that their confidentiality and privacy would be maintained to the fullest extent possible and that any identifying details will only be published with their explicit permission. They were brief about their right to withdraw their consent at any time before the publication process is completed, without any punishment to their participation in the research. Availibiliy of data and material Data and materials used in this research are not available for sharing due to privacy concerns protecting individuals confidentiality and privacy, since it is involving human subjects, which might contain sensitive information. Materials are no longer available due to resource constraints or supply chain disruptions Acknowledgements Thanks to Dr. A.G. Ogofure and the members of staff of Department of Microbiology, University of Benin, Benin City, Nigeria for their enabling environment for the Laboratory work. The authors wish to also acknowledge support received from TETFUND –Centre of Excellecne for Biomedical, Engineering and Agricultural Translational studies UNIZIK, Awka, Nigeria. References Adegoke, A., Faleye, A., Singh, G. and Stenström, T. (2016). Antibiotic-Resistant Superbugs: Assessment of the Interrelationship of Occurrence in Clinical Settings and Environmental Niches. Molecules: A Journal of Synthetic Chemistry and Natural Product Chemistry 22 . https://doi.org/10.3390/molecules22010029. Bacci, S., Mølbak, K., Kjeldsen, M.K. and Olsen, K.E. (2011). Binary toxin and death after Clostridium difficile infection. Emerging Infectious Diseases 17 :976–982. Barbut F., P. Mastrantonio, M. Delme´e, J. Brazier, E. Kuijper, and I.Poxton ( 2007). Prospective study of Clostridium difficile -associated disease in Europe with phenotypic and genotypic characterization of the isolates. Clin. Microbiol. Infect. 13: 1048–1057. Bridson, E.Y. (2006). The Oxoid Manual (9 th Edition). Oxoid Limited, Basingstoke Hamsphire, England 623 pp. Buddle, J. E. and Fagan, R. P. (2023). Pathogenicity and virulence of Clostridioides difficile . Virulence 14 (1): 2150452. Call, D.R., Bakko, M.K., Krug, M.J. and Roberts, M.C. (2003). Identifying antimicrobial resistance genes with DNA microarrays. Antimicrobial Agents and Chemotherapy 47 : 3290 – 3295. 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A multicentre analysis of Clostridium difficile in persons with Cystic Fibrosis demonstrates that carriage may be transient and highly variable with respect to strain and level: Cystic Fibrosis and Clostridium difficile. The Journal of Infection . https://doi.org/10.1016/j.jinf.2020.12.027. Doyle, E.M. (2013). Clostridium difficile as a Risk Associated with Animal Sources. FRI Food Safety Reviews. Food Research Institute, University of Wisconsin, pp. 1-18. Flórez, A.B., Alegría, Á., Rossi, F., Delgado, S., Felis, G.E., Torriani, S. and Mayo, B. (2014). Molecular identification and quantification of tetracycline and erythromycin resistance genes in Spanish and Italian retail cheeses. BioMed Research International. https://doi.org/10.1155/2014/746859 Freeman J., W. N. Fawley, J. Hobbs, P. Else, P. Verity, P. Parnell, S., D., Baines, and M. H. Wilcox ( 2009). Decreasing metronidazole susceptibility in epidemic Clostridium difficile strains, abstr. C2-1964. Abstract on 49th Interscience Conference on Antimicrobial Agents and Chemotherapy, San Francisco, CA. Gerding, D. N. ( 2004). Clindamycin, cephalosporins, fluoroquinolones, and Clostridium difficile -associated diarrhea: this is an antimicrobial resistance problem. Clin. Infect. Dis . 38: 646–648. Jacek, C., Mirosław, D., Hanna, P., Kuijper, J., William, P., Aleksandra, M., Sarah, G., Dorota, W., Aleksander, G., and Grażyna, B., (2019). European Journal of Clinical Microbiology and Infectious Diseases . 38 (7): 1211–1221 Jobstl, M., Heuberger, S., Indra, A., Nepf, R., Kofer, J. and Wagner, M. (2010). Clostridium difficile in raw products of animal origin. International Journal of Food Microbiology. 138 (2):172–175. Johnson, S., Kent, S.A. and O’Leary, K.J. (2001). Fatal pseudomembranous colitis associated with a variant Clostridium difficile strain not detected by toxin A immunoassay. Annals of Internal Medicine. 135 : 434–438. Khoder, G., Muhammad, J. S., Mahmoud, I., Soliman, S. S. and Burucoa, C. (2019). Prevalence of Helicobacter pylori and its associated factors among healthy asymptomatic residents in the United Arab Emirates. Pathogens 8( 2 ): 44. Kuehne, S., Cartman, S., Heap, J., Kelly, M., Cockayne, A., & Minton, N. (2010). The role of toxin A and toxin B in Clostridium difficile infection. Nature , 467, 711-713. https://doi.org/10.1038/nature09397. Lawley, T.D., Croucher, N.J., Yu, L.U., Clare, S., Sebaihia, M., Goulding, D., Pickard, D.J., Parkhill, J., Choudhary, J. and Dougan, G. (2009). Proteomic and genomic characterization of highly infectious Clostridium difficile 630 spores . Journal of Bacteriology. 191 :5377–5386. Markantonis, J.E., Fallon, J.T., Madan, R. and Alam, M.Z. (2024). Clostridioides difficile Infection: Diagnosis and Treatment Challenges. Pathogens 13 : 118. https://doi.org/10.3390/pathogens13020118 Martirosian, G., Polanski, J., Szubert, A. and Meisel-Mikolajczyk, F. (1993). Clostridium difficile in a department of surgery. Materia medica Polona 25 :45-47. Mondal, S. (2019). Prospective observational study to assess patient profile with Clostridium difficile colitis. Mori, N. and Aoki, Y. (2015). Clinical characteristics and risk factors for community-acquired Clostridium difficile infection: A retrospective, case-control study in a tertiary care hospital in Japan. Journal of Infection and Chemotherapy: Official Journal of the Japan Society of Chemotherapy 21 (12): 864-867. https://doi.org/10.1016/j.jiac.2015.09.004. Panhwar, A., Abro, R., Kandhro, A., Khaskheli, A. R., Jalbani, N., Gishkori, K. A. and Qaisar, S. (2022). Global Water Mapping, Requirements, and Concerns over Water Quality Shortages. Parija, S. C. (2023). Clostridium. In Textbook of Microbiology and Immunology (pp. 465-488). Singapore: Springer Nature Singapore. Rodriguez-Palacios, A., Ilic, S. and LeJeune, J.T (2014). Clostridium difficile with Moxifloxacin/Clindamycin Resistance in Vegetables in Ohio, USA, and Prevalence Meta-Analysis. Journal of Pathogens 7 Rodriguez-Palacios, A., Staempfli, H. R., Duffield, T., Weese, J. S. (2007). Clostridium difficile in retail ground meat, Canada. Emerging Infectious Diseases. 13 (3):485–487. Rupnik, M., Wilcox, M.H. and Gerding, D.N. (2009). Clostridium difficile infection: new developments in epidemiology and pathogenesis. Nature Reviews Microbiology 7 :526–536. Sheen, E., Pan, J., Ho, A.andTriadafilopoulos, G. (2021). Lower Gastrointestinal Bleeding. In Geriatric Gastroenterology (pp. 1305-1325). Cham: Springer International Publishing. Shehabi A. A., Abu-Ragheb H. A. and Allaham N. A. (2001). Prevalence of Clostridium difficile -associated diarrhoea among hospitalized Jordanian patients. Eastern Medical and Health Journal. 7 :750-5. Shokoohizadeh, L., Alvandi, F., Yadegar, A., Azimirad, M., Hamid-Hashemi, S. and Alikhani, M.A. (2021). Frequency of toxin genes and antibiotic resistance pattern of Clostridioides difficile isolates in diarrheal samples among hospitalized. patients in Hamadan, Iran. GastroenterolHepatol Bed Bench 14(2):165-173 Simango, C. and Mwakurudza, S. (2008). Clostridium difficile in broiler chickens sold at marketplaces in Zimbabwe and their antimicrobial susceptibility . International Journal of Food Microbiology. 124 : 268–270. Voutsinas, N., Toussie, D., Jacobi, A., Bernheim, A.and Chung, M. (2020). Incidental CT findings in the lungs in COVID-19 patients presenting with abdominal pain. Clinical Imaging 67: 1-4. Warriner, K., Xu, C., Habash, M., Sultan, S. and Weese, S. (2017). Dissemination of Clostridium difficile in food and the environment: Significant sources of C. difficile community‐acquired infection?. Journal of Applied Microbiology 122 . https://doi.org/10.1111/jam.13338. Weese, J.S., Finley, R., Reid-Smith, R. R., Janecko, N. and Rousseau, J. (2010). Evaluation of Clostridium difficile in dogs and the household environment. Epidemiological Infection. 138 :1100–1104. Willey, L.M., Sherwood, L.J. and Woolverton, L. (2008). Microbiology (7 th Edition). Mc-Graw Hill, New York, 966 pp. Xiaolu, L., Yizhong, W., Rong, C., Fangfei, X., Xufei, W., Lin, Y., Yongmei, X., Dan, L. and Ting, Z. (2024). Antimicrobial Resistance of Clostridioides difficile in Children from a Tertiary Pediatric Hospital in Shanghai, China. Infection and Drug Resistance 17 : 329–339. Plate Plate 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Plate1.docx Cite Share Download PDF Status: Posted Version 1 posted 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. 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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-4955956","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":352537470,"identity":"aaf83ff2-6b1c-4628-85eb-fe8339a5bf5a","order_by":0,"name":"Eddison Igho Oghonyon","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA90lEQVRIiWNgGAWjYFACHijN3mDAUMFwAMQ0IFILzwEDhjOkaZFIIFILP//ag48L22zy+CUfb5M42HbHnoG9eZsE445anFokZ7xLNp7ZllYsOTutDKjlWWIDz7EyCcYzx3FqMbhxxkyat+1w4obbOWbSH9sOJzBI5JhJMLYdI6xl/80zZkBbDtszyL8hoOV8D9QWCR6wFsYGEIOxrQaPX/iSjXnOpSXOOJNWbHHg3OHENh4gI7HtAE4t/PxnDz7mKbNJ7G8/vPHGgbLD9vzsQMbHtjqcWoDRwcDAyIYkAGYnMBzGrYUf5II/mOJ4bBkFo2AUjIKRBgD/CFoWDzh7GgAAAABJRU5ErkJggg==","orcid":"","institution":"Nnamdi Azikiwe University","correspondingAuthor":true,"prefix":"","firstName":"Eddison","middleName":"Igho","lastName":"Oghonyon","suffix":""},{"id":352537471,"identity":"af9370c1-6024-4173-8548-6c8833098757","order_by":1,"name":"Malachy C. Ugwu","email":"","orcid":"","institution":"Nnamdi Azikiwe University","correspondingAuthor":false,"prefix":"","firstName":"Malachy","middleName":"C.","lastName":"Ugwu","suffix":""},{"id":352537472,"identity":"c3c6821e-317d-48e8-93b2-303b29e10484","order_by":2,"name":"Charles O. Esimone","email":"","orcid":"","institution":"Nnamdi Azikiwe University","correspondingAuthor":false,"prefix":"","firstName":"Charles","middleName":"O.","lastName":"Esimone","suffix":""},{"id":352537473,"identity":"00ec80cc-a648-4688-96ef-48e7eac602bf","order_by":3,"name":"Anthony Ik Onah","email":"","orcid":"","institution":"David Uhami Federal University of Health Sciences, Uburu Ebonyi State, Nigeria","correspondingAuthor":false,"prefix":"","firstName":"Anthony","middleName":"Ik","lastName":"Onah","suffix":""}],"badges":[],"createdAt":"2024-08-22 07:24:34","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4955956/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4955956/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":72560934,"identity":"dbcd7f81-5f95-4ec1-99f1-306fba25a226","added_by":"auto","created_at":"2024-12-29 22:16:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1276719,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4955956/v1/3b367d3d-7c9f-40b7-9737-59483a324e91.pdf"},{"id":66041325,"identity":"7d3ce925-62a0-4f0d-8ca1-e94b705fc8fb","added_by":"auto","created_at":"2024-10-07 05:58:40","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":142049,"visible":true,"origin":"","legend":"","description":"","filename":"Plate1.docx","url":"https://assets-eu.researchsquare.com/files/rs-4955956/v1/38cef4c10bad4860c5b6b068.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003ePrevalence, Molecular Characterization, Pathogenicity and Antimicrobial Susceptibility Pattern of Clostridioides Difficile From Clinical Samples in South Eastern Nigeria.\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eStandard healthcare facilities are essential for survival in a world dominated by superbugs,which have also been implicated in infections and the re-occurrence of infection even after the treatment regimen. Pathogenic organisms have recorded unparalleled success in their ability to cause diseases as well as induce therapeutic failure through drug resistance (Adegokeet al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003cem\u003eClostridioides difficile\u003c/em\u003e has been reported as the leading cause of healthcare-associated diarrhea (Markantonis et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Antibiotic exposure, which reduces the population of non-pathogenic anaerobes that normally inhabit the gut, has been pointed out asthe most important risk factor for \u003cem\u003eClostridioides difficile\u003c/em\u003e infection (CDI) (Markantonis et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).Thus, CDI occurs when there is a shift in the microbial flora of the colon, allowingtoxin-producing strains of the Gram-positive, spore-forming, anaerobic bacillus toproliferate (Smits et al., 2016; Markantonis et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Other events implicated in gut microbiota disruption includeproton pump inhibitors (PPI) use and inflammatory bowel diseases (Buddle and Fagan, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Xiaolu et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Researchers have attributed the pathogenicity of \u003cem\u003eC. difficile\u003c/em\u003e to the two major virulence factors produced by toxigenic \u003cem\u003eC. difficile\u003c/em\u003e strains, enterotoxin A (\u003cem\u003eTcdA\u003c/em\u003e) and cytotoxin B (\u003cem\u003eTcdB\u003c/em\u003e) (Buddle and Fagan \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Xiaolu et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eOn September 16th, 2013, the Centre for Disease Control (CDC) declared \u003cem\u003eClostridioides difficile\u003c/em\u003e as a threat with \u0026ldquo;urgent\u0026rdquo; concern to public health in the United States due to the identification of antimicrobial resistance among human and animal-derived \u003cem\u003eClostridioides difficile\u003c/em\u003e isolates to drugs commonly used in humans and also to their increased rates of associated patient mortality since the early 2000s(CDC, 2013).\u003c/p\u003e \u003cp\u003eIn all the studies, some strains of \u003cem\u003eC. difficile\u003c/em\u003e appear to be species-specific, while other strains were isolated from multiple species, including man. The organism was initially described as part of the normal flora in stool samples from healthy infants in 1935 (Parija, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), as well as its detection in significant numbers in healthy asymptomatic children (Khoder et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). It was afterwardidentified as a pathogen associated with several diseases, and even now, in hospitals and long-term care facilities, it has become the most common cause of diarrhea, causing the loss of billions of dollars in healthcarecosts (Panhwar, 2022).The symptoms of infection usually appear mild in some cases, and in others, they appear to be life-threatening or fatal(Mondal, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Diarrhea associated with \u003cem\u003eC. difficile\u003c/em\u003e is accompanied by the passage of occult blood or mucus in the stool, but hematochezia and melena are other rare symptoms (Sheen et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Xiaoluet al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Certain other symptoms are found in less than 50% of infected patients, including abdominal discomfort, fever, and peripheral leukocytosis (Voutsinaset al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).Disease caused by \u003cem\u003eC. difficile\u003c/em\u003e is linked most commonly to nosocomial infections in humans, especially after empiric antibiotic treatment. Humans develop a spectrum of diseases when infected with some strains of \u003cem\u003eC. difficile\u003c/em\u003e. It has been shown that not all people who are on prolonged antibiotic medication, as well as those above the age of 60 years, are susceptible to CDI(Jaceket.al, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The advent of some virulent new strains of \u003cem\u003eC. difficile\u003c/em\u003e has triggered an increase in CDI rates even in younger populations originally thought to be at low risk of contracting \u003cem\u003eC. difficile\u003c/em\u003e infection (Doyle, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).To date, little is known about CDI in Southeastern Nigeria as inadequate attention has been given to the understanding of the molecular epidemiology of CDI in Nigeria, a high-burden country.Thus, the research was aimedatdeterminingthe prevalence, molecular characterization, pathogenicity, and antimicrobial susceptibility pattern of \u003cem\u003eClostridioides difficile\u003c/em\u003e from clinical samples in the South-Eastern states of Nigeria.\u003c/p\u003e"},{"header":"METHOD","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy Area\u003c/h2\u003e \u003cp\u003e The study was conducted after obtaining due ethical approval from the ethical committee of the ChukwuemekaOdumegwuOjukwu University Teaching Hospital (COUTH), Amaku, Awka,for the collection of stool samples in the hospital in Awka andneighbouring states.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSample Collection\u003c/h3\u003e\n\u003cp\u003eThe clinical samples were collected from selectedpublic hospitals.Stool samples were collected in red cap universal bottles from COOUTH in Anambra, Federal Teaching Hospital Abakaliki in Ebonyi, Abia State University Teaching Hospital in Abia, Imo State University Teaching Hospital in Imo and Enugu State Teaching Hospital in Enugu. Samples were collected from patients for analysis in the clinical laboratory, after which they werecollected and preserved in an ice pack for further analysis. The samples were collected and someof the challeges in collection includes: untimely collection, patient unwillingness to cooperate, cross contamination risk,health and safety issues A total of 450 stool samples were collected and evaluated in this study. The sample size was determined using the sample size determination formula below:\u003c/p\u003e \u003cp\u003e \u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(\\:Samplesize\\:\\left(N\\right)=\\:{(Z}^{2\\:}P\\left(1-P\\right))\u0026divide;{D}^{2}\\)\u003c/span\u003e \u003c/span\u003e \u0026hellip;\u0026hellip;\u0026hellip;\u0026hellip;.Eq.\u0026nbsp;1\u003c/p\u003e \u003cp\u003ewhere: \u003cem\u003eN\u0026thinsp;=\u0026thinsp;Sample size, Z\u0026thinsp;=\u0026thinsp;z score (1.969), P\u0026thinsp;=\u0026thinsp;prevalence (0.45) andD\u0026thinsp;=\u0026thinsp;0.05\u003c/em\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003eIsolation and Identification of\u003c/b\u003e \u003cb\u003eClostridium difficile\u003c/b\u003e\u003c/p\u003e \u003cp\u003eFor the isolation of \u003cem\u003eClostridioides difficile\u003c/em\u003e, all samples were pre-treated (heat shocked and alcohol shocked) to reduce competing flora and allow the growth of spore-forming bacilli. AStandard microbiological procedure was employed using \u003cem\u003eClostridium difficile\u003c/em\u003e differential agar CM 0601 (Oxoid) supplemented with \u003cem\u003eClostridium difficile\u003c/em\u003e selective supplement SR0096 (Oxoid) (Bridson, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Stock solutions were prepared for the samples by weighing 20g of the stool samples into 180 ml of sterile saline water in a conical flask. A ten-fold serial dilution from the stock solution was made after mixing. From the fifth tube, 2 ml of the suspension was added to a well-labelled tube containing 2 ml of absolute ethanol (alcohol/ethanol shock method). The mixture was rocked gently to allow for homogenization, and it was left to stand for an hour before plating (of 100\u0026micro;l) was carried out. Heat treatment, also known as heat shocking, was employed by heating the tubes containing a mixture of samples to boil for 5 minutes. The tubes were then allowed to cool before plating was carried out on the media supplemented with \u003cem\u003eClostridium difficile\u003c/em\u003e supplement (Oxoid).The formula employed for the dilution factor is given below in Eq.\u0026nbsp;(2)\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:Dilution\\:factor\\:=\\:\\frac{final\\:volume}{aliquot\\:volume\\:}\\left(2\\right)$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\:where:final\\:volume=aliquot\\:volume\\:\\left(sample\\:volume\\right)\\:+diluent\\:volume$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eEnumeration of the bacterial/fungal isolates was carried out using the formula delineated by Willey et al. (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), and it is shown in Eq.\u0026nbsp;(3) below.\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\n$$\\:\\frac{cfu}{g}=\\frac{number\\:of\\:colonies\\:x\\:dilution\\:factor}{volume\\:of\\:inoculum\\:}\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\left(3\\right)\\:$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eThe cultural, morphological, microscopic and biochemical properties of the isolates were evaluated after Gram and spore staining was done to select the positive isolates for both stains.\u003c/p\u003e \u003cp\u003eBiochemical tests like catalase, oxidase, indole, urease and sugar fermentation tests were carried out to identify \u003cem\u003eClostridium\u003c/em\u003e species. The identity of \u003cem\u003eClostridioides\u003c/em\u003e difficile was finalized following blasting of sequences obtained using 16sRNA universal primer sets.\u003c/p\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eAntibiotic Susceptibility Test\u003c/h2\u003e \u003cp\u003eThe identified \u003cem\u003eC. difficile\u003c/em\u003e were subjected to standard antibacterial susceptibility testing (AST). The antibiotic discs (Oxoid UK) containing the following antibiotics Meropenem (10 \u0026micro;g), Erythromycin (15 \u0026micro;g), Metronidazole (50 \u0026micro;g), Amoxicillin/clavulanic acid (20/10 \u0026micro;g), Clindamycin (20 \u0026micro;g), Gentamicin (10 \u0026micro;g), Ciprofloxacin (5 \u0026micro;g), Vancomycin (30 \u0026micro;g), and Tetracycline (30 \u0026micro;g) was used. Briefly, an18-24 hours bacterial culture wascultured on MHA. The inoculum corresponding to 1.5 x 10\u003csup\u003e8\u003c/sup\u003e cells/ml McFarland standard wasstreaked using a sterile loop onto the MHA plates before the antibiotic discs were aseptically added with extreme care to the plates with the aid of sterile forceps. The susceptibility results were recorded after incubation for 24 hours at 37 \u0026ordm;C and interpreted usingClinical Laboratory Standards Institute breakpoints (CLSI, 2020).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eMultiple Antibiotic Resistance Index determination\u003c/h2\u003e \u003cp\u003eThe MAR index was determined by the formula by Chitanandet al. (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2010\u003c/span\u003e):\u003c/p\u003e \u003cp\u003e \u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(\\:MARindex=\\frac{y}{nx}\\)\u003c/span\u003e \u003c/span\u003e\u0026hellip;\u0026hellip;\u0026hellip;..Eq.\u0026nbsp;4\u003cdiv id=\"Equd\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equd\" name=\"EquationSource\"\u003e\n$$\\:wherey=numberofresistancescored,\\:n=numberofisolates$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Eque\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Eque\" name=\"EquationSource\"\u003e\n$$\\:x=total\\:number\\:of\\:antibioitics$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eExtraction of DNA\u003c/h2\u003e \u003cp\u003eDNA was extracted using boiling method described by Chakravorty \u003cem\u003eet al\u003c/em\u003e. (2007). 2 ml of 24 h. broth culture of \u003cem\u003eC. difficile\u003c/em\u003e was transferred to tube and centrifuged at high speed for 5 min. The supernatant was discarded after which 200 \u0026micro;L of sterile distilled water (SDW) was added to the pellets and vortexed using a vortex mixer for 1 min before it was heated at 100 \u003csup\u003eo\u003c/sup\u003eC for 15 mins. Lastly, it was thereafter centrifuge at high speed for 2 minutes. The supernatant from the second spin was now regarded as the pure DNA and from which, 10 \u0026micro;L of DNA was used for amplification of the gene by PCR.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrimer sets for virulence genes\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGenes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePrimer\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSequence (5\u0026rsquo;-3\u0026rsquo;)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eConc. (\u0026micro;M)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eProduct size (bp)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTcdA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003etcdA-F\u003c/p\u003e \u003cp\u003etcdA-R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGTATGGATAGGTGGAGAAGTCAGTG\u003c/p\u003e \u003cp\u003eCGGTCTAGTCCAATAGAGCTAGGTC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e632\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003etcdB\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003etcdB-F\u003c/p\u003e \u003cp\u003etcdB-R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGAAGATTTAGGAAATGAAGAAGGTGA\u003c/p\u003e \u003cp\u003eAACCACTATATTCAACTGCTTGTCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e441\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ecdtA\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecdtA-F\u003c/p\u003e \u003cp\u003ecdtA-R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eATGCACAAGACTTACAAAGCTATAGTG\u003c/p\u003e \u003cp\u003eCGAGAATTTGCTTCTATTTGATAATC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e260\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ecdtB\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecdtB-F\u003c/p\u003e \u003cp\u003ecdtB-R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eATTGGCAATAATCTATCTCCTGGA\u003c/p\u003e \u003cp\u003eCCAAAATTTCCACTTACTTGTGTTG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e179\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe presence of genes which code for virulence (\u003cem\u003etcdA, tcdB, cdtA\u003c/em\u003e and \u003cem\u003ecdtB\u003c/em\u003e) was determined using standardized primer sets in the table above.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrimer sets for resistance genes\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGenes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePrimer\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSequence (5\u0026rsquo;-3\u0026rsquo;)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eConc. (\u0026micro;M)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eProduct size (bp)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eerm(B\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eErmB-F\u003c/p\u003e \u003cp\u003eErmB-R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGAAAAGGTACTCAACCAAATA\u003c/p\u003e \u003cp\u003eAGTAACGGTACTTAAATTGTTTAC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e639\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003etet(S)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTetS-F\u003c/p\u003e \u003cp\u003eTetS-R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eATCAAGATATTAAGGAC\u003c/p\u003e \u003cp\u003eTTCTCTATGTGGTAATC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e573\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003etet(A)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003etet(A)- F\u003c/p\u003e \u003cp\u003etet(A)- R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTTGGCATTCTGCATTCACTC\u003c/p\u003e \u003cp\u003eGTATAGCTTGCCGGAAGTCG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e494\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe genes coding for erythromycin (\u003cem\u003eerm\u003c/em\u003eB) and tetracycline resistance (\u003cem\u003etet\u003c/em\u003eA and \u003cem\u003etetS\u003c/em\u003e) were screened using polymerase chain reaction (PCR). The previously extracted bacterial DNA sample was used for the analysis. Agarose gel electrophoresis was carried out using each primer (forward and reverse primers) for the target genes (Call et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Florez \u003cem\u003eet al\u003c/em\u003e., 2014). A thermocycler was used to determine an optimal annealing temperature for the specific binding of the primer set to the DNA template.\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTotal anaerobic bacterial count of stool samples obtained from patients in different states\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLocation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of Samples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAnaerobe count (Log\u003csub\u003e10\u003c/sub\u003eCFU/g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003ep-value\u003c/em\u003e(s)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnambra\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.168\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEbonyi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.168\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEnugu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.168\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eImo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.066\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAbia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.066\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe results of the anaerobic stool sample count from Eastern Nigeria areshown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e above.The total anaerobe count (log\u003csub\u003e10\u003c/sub\u003eCFU/g) for stool samples obtained from patients in different states showed that stool samples obtained from patients in Anambra had counts of 5.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09 log\u003csub\u003e10\u003c/sub\u003eCFU/g. In contrast, samples from Ebonyi, Enugu and Imo states had counts of 5.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 log\u003csub\u003e10\u003c/sub\u003eCFU/g, 5.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 log\u003csub\u003e10\u003c/sub\u003eCFU/g and 5.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 log\u003csub\u003e10\u003c/sub\u003eCFU/g, respectively.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003ePlate 1. Agarose gel electrophoresis of bacterial DNA isolated from stool samples\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eIdentity of bacterial isolates (anaerobes) from stool samples from Eastern Nigeria\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSample Code\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSource\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIdentity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eQuery Cover(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eHomology (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAccession number\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanStool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e100.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMH888201.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eebstool05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e91.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP019858.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eImstool15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP019860.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanStool21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP025047.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanStool22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e90.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP020424.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabstool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e100.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMH888202.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabstool17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP019860.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eimstool83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e94.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP028361.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eimstool49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e94.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP028361.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eebstool23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e94.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP028361.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eebstool33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP019860.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eebstool1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP025047.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabstool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e90.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCP020424.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanstool70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eClostridioides difficile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e100.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMH888202.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe identity and Phylogenetic relatedness of anaerobic bacterial isolates obtained from stool samples in Eastern Nigeria are shown in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e above.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrevalence of anaerobes from stool samples using standard cultural methods\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSamples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSamples Number\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAnambra\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEbonyi\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eEnugu\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eImo\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAbia\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTotal Sample(study)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eStudy Total (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStool Samples\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55(61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e45(50)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50(55)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e41(46)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50(55)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e450\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e241(54)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe prevalence of anaerobes from stool samples using standard cultural methods (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e) revealed that all stool samples but Imo (46%) had anaerobic counts equal to or greater than 50% following a cultural phenotypic standard. In comparison, a total of 54% was obtained in the entire study.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrevalence of \u003cem\u003eClostridioides difficile\u003c/em\u003e from stool samples in the study\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSamples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSample Number\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAnambra\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEbonyi\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eEnugu\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eImo\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAbia\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTotal sample (study)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eStudy Total (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStool Samples\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4(44.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2(22.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2(22.22)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3(33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e450\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e14(3.11)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe prevalence of \u003cem\u003eClostridioides difficile\u003c/em\u003e from stool samples (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e) revealed a total study prevalence of 3.11% for stool samples. Stool samples from Ebonyi and Enugu states had a prevalence of 2.22% apiece. In comparison, Imo and Abia states had a 3.33% prevalence of \u003cem\u003eC. difficile\u003c/em\u003e from stool samples, with Anambra state having the highest prevalence of 4.44%.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePercentage Recovery of \u003cem\u003eC. difficile\u003c/em\u003e from stool samples using cultural methods\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSample\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHeat Treatment (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAlcohol Treatment (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eStudy Total (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStool Samples\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e450\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5(1.11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14(3.11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14(3.11)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe prevalence of \u003cem\u003eC. difficile\u003c/em\u003e recovered by the heat treatment method (1.11%) was found to be lower than the percentage recovered via alcohol treatment (3.11%).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ea: Antibiotic sensitivity profile to common antibiotics\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"11\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCode\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSamples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMEM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eERY\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMET\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAMC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCL\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eCIP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eVA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eTET\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanStool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eebStool05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eimStool15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanStool21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanStool22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabStool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabStool77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eimStool83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eimstool49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eebstool23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eenstool33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eenstool1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabStool14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanstool70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"11\"\u003eKeys: Meropenem (10 \u0026micro;g), Erythromycin (15 \u0026micro;g), Metronidazole (50 \u0026micro;g), Amoxicillin/clavulanic acid (20/10 \u0026micro;g), Clindamycin (20 \u0026micro;g), Gentamicin (10 \u0026micro;g), Ciprofloxacin (5 \u0026micro;g), Vancomycin (30 \u0026micro;g), Tetracycline (30 \u0026micro;g)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAntibacterial sensitivity pattern and percentage of resistance/susceptibility of \u003cem\u003eC. difficile\u003c/em\u003e isolates stool samples to common antibiotics showed susceptibility to carbapenems (82.61%), aminoglycosides (86.98%) and Β-Lactam/combination agents (73.91%). Notable resistance was observed for tetracycline (73.91%), macrolides (73.91%) and fluoroquinolones (43.48%), respectively.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab9\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eb: Percentage of antibiotic resistance and susceptibility profile of common antibiotics\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAntibiotics Classes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAntibiotics\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eSusceptibility profile \u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;09\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSensitive (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eResistant (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCarbapenems\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMEM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19(82.61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3(17.39)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMacrolides\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eERY\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6(26.09)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e17(73.91)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNitroimidazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMET\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16(69.56)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7(30.43)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eΒ-Lactam/combination agents\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAMC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e17(73.91)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6(26.09)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLincosamide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15(65.22)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8(34.78)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAminoglycosides\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e20(86.98)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3(13.02)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFluoroquinolone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCIP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13(56.52)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10(43.48)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGlycopeptides\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eVA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16(69.56)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7(30.43)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTetracyclines\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTET\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6(26.09)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e17(73.91)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eKey: Meropenem (10 \u0026micro;g), Erythromycin (15 \u0026micro;g), Metronidazole (50 \u0026micro;g), Amoxicillin/clavulanic acid (20/10 \u0026micro;g), Clindamycin (20 \u0026micro;g), Gentamicin (10 \u0026micro;g), Ciprofloxacin (5 \u0026micro;g), Vancomycin (30 \u0026micro;g), Tetracycline (30 \u0026micro;g).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab10\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 9\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMultiple antibiotic resistance distribution of \u003cem\u003eC. difficile\u003c/em\u003e isolates from clinical sources\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIsolate Code\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAntibacterial Classes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNumber of Antibiotics\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eResistance Phenotypes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eResistant phenotypes (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMAR Index\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanBE7, ebstool05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAMCR, VAR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2(8.69)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eenLV4, ebstool23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMEMR, ERYR, METR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2(8.69)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eebLV5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eERYR, CNR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanStool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eERYR, AMCR, VAR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eimstool15, imstool49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eERYR, METR, CLR, CIPR, TETR,\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2(8.69)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanstool21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanstool222\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eERYR, CIPR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabstool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMEMR, ERYR, METR, CLR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eenstool33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eERYR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eenstool1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eERYR, CLR, CIPR, TETR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eabstool14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAMCR, VAR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eanstool70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCIPR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1(4.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003e\u003cb\u003eKeys\u003c/b\u003e: Meropenem (10 \u0026micro;g), Erythromycin (15 \u0026micro;g), Metronidazole (50 \u0026micro;g), Amoxicillin/clavulanic acid (20/10 \u0026micro;g), Clindamycin (20 \u0026micro;g), Gentamicin (10 \u0026micro;g), Ciprofloxacin (5 \u0026micro;g), Vancomycin (30 \u0026micro;g), Tetracycline (30 \u0026micro;g)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe multiple antibiotic resistance phenotypic distribution of \u003cem\u003eC. difficile\u003c/em\u003e isolates from clinical samples (Table\u0026nbsp;\u003cspan refid=\"Tab10\" class=\"InternalRef\"\u003e9\u003c/span\u003e) showed that isolates with a MAR index of 0.44 had the highest resistance phenotypes. In contrast, isolates with a MAR index of 0.33 had the next highest resistance phenotypes.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab11\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 10\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrevalence of virulence genes in isolates of \u003cem\u003eC. difficile\u003c/em\u003e obtained from stool samples\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLanes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCode\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSamples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003etcdA\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003etcdB\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003ecdtA\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003ecdtB\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTotal (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanStool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eebstool05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eImstool15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanstool21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1(25)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanstool22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eabstool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eabstool77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eimstool83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1(25)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eimstool49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eebstool23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eenstool33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eenstool11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1(25)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eabstool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanstool70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003csub\u003e+\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2(50)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e \u003cp\u003eTotal Study(%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16(69.56)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e23(100.00)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1(4.34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1(4.34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAll (100%) of \u003cem\u003eC. difficile\u003c/em\u003e isolates were found to possess \u003cem\u003etcdB\u003c/em\u003egenes. About \u003cem\u003etcdA\u003c/em\u003egenes, 10 of 14 positive isolates were found to possess the virulence genes, making a total of 69.56% of isolates with positive results for the presence of the genes. None of the isolated \u003cem\u003eC. difficile\u003c/em\u003e strains but one possessed \u003cem\u003ecdtA\u003c/em\u003eand \u003cem\u003ecdtB\u003c/em\u003e genes, thus giving a percentage of 4.34% apiece for the aforementioned genes. The prevalence of virulence genes in Table\u0026nbsp;\u003cspan refid=\"Tab11\" class=\"InternalRef\"\u003e10\u003c/span\u003e revealed that 3 isolates possessed 25% (one of the four) of virulence genes analyzed in the study, while 10 isolates possessed two of the four virulence genes (50%)\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab12\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 11\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrevalence of resistant genes to erythromycin and tetracycline found in isolates of \u003cem\u003eC. difficile\u003c/em\u003e obtained from stool samples\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLanes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCode\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSamples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eTetA\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003etetS\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eErmB\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eTotal(%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanStool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eebstool05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2(67)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eImstool15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanstool21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2(67)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanstool22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eabstool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eabstool77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eimstool83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1(33)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eimstool49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eebstool23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eenstool33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eenstool1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3(100)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eabstool12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0(0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eanstool70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStool\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0(0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e \u003cp\u003eTotal study (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17(73.91)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17(73.91)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17(73.91)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e. Agarose gel electrophoresis of resistance genes present in \u003cem\u003eC. difficile\u003c/em\u003e obtained from stool samples is shown in Table\u0026nbsp;\u003cspan refid=\"Tab12\" class=\"InternalRef\"\u003e11\u003c/span\u003e. In contrast,\u003cem\u003eerm(B)\u003c/em\u003egene PCR products amplified from \u003cem\u003eClostridioides difficile\u003c/em\u003e isolates were found in 73.91%; it was also the same percentage that was present for tetracycline resistance (\u003cem\u003etet(S)\u003c/em\u003e and \u003cem\u003etet(A)\u003c/em\u003e genes). Nine isolates were found to possess all resistance genes analyzed in the study\u003c/p\u003e \u003cp\u003e.\u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThe majority of bacteria (anaerobes) in the stool are commensals, which oftendo not harm humans unless there is an underlying risk factor or the case of a compromised immune system. The bacterium of interest in this study (\u003cem\u003eC. difficile\u003c/em\u003e)\u0026nbsp;is regarded as both a commensal organism and a pathogen to domestic animals (Doyle, 2013). Most of these anaerobes in the gut that have been isolated from the feces are not competitive, and their population can only explode, or they can thrive better in the gut after an array of antibiotic treatments which could destroy much of the intestinal normal flora (Doyle, 2013). The statistical comparison of the total anaerobic count in stool samples showedno significant difference (\u003cem\u003ep\u0026gt;0.05\u003c/em\u003e) between samples obtained from Ebonyi and other Eastern states in the study. However, there was a significant difference between samples obtained from Anambra and those from Imo and Abia states, respectively.\u003c/p\u003e\n\u003cp\u003eThe bacterial identity, query cover, and percentage homology were written against the confirmed blasted identity of the isolates,including accession numbers obtained from the National Centre for Biotechnology Information (NCBI) website. The 3.11% prevalence of \u003cem\u003eClostridiumdifficile\u0026nbsp;\u003c/em\u003eobtained in this study is less than several reported prevalences in the literature. This implies that the probability of infection due to \u003cem\u003eC.\u0026nbsp;\u003c/em\u003edifficile\u0026nbsp;would be very low.However, the presence of the pathogen (even in its low prevalence) in this study is an indication that there is a likely chance of a CDI outbreak if proper measures are not in place to curtail the infection. Following reports by Warriner\u003cem\u003eet al\u003c/em\u003e. (2017),\u0026nbsp;\u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003ewas previously thought to be clinically associated alone. Still, with advanced scientific methods, it is now being muted as a community-acquired infection. The prolonged survival of spores of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003ein the environment might also increase the possibility of animal and food contamination which is a potential source of infection(Weese, 2010). In Europe, a higher prevalence had also been reported thanthe one obtained in this study, howbeit for hospitalized patients with even higher reports of toxigenic strains of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003erecovered from patients’stool\u0026nbsp;(Martirosian\u003cem\u003eet al\u003c/em\u003e., 1993; Shehabi\u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2001). Nonetheless, findings in this study corroborated the report of Mori and Aoki (2015) and Deane \u003cem\u003eet al.\u003c/em\u003e (2021),who opined that carriage rates of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003ein healthy adults varied from 0% to 3% in Europe to up to 15% in Japan.\u003c/p\u003e\n\u003cp\u003eFollowing cultural phenotypic methods, the percentage recovery of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003efrom stool samples revealed that while both treatment methods (heat and alcohol treatments) yielded positive \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003egrowth, the alcohol shock method was found to be better at recovering the bacterium isolates from clinical samples. The findings in this study about how the spores of the bacterium were recovered from both treatment processes agreed with the findings of Lawley \u003cem\u003eet al\u003c/em\u003e. (2009).\u0026nbsp;They reported that spores are difficult to eradicate due to their ability to resist radiation, heat and chemicals (such as alcohols), which are usually employed as disinfectants in both hospital and community settings. In this study, alcohol shock samples (3.11%) had a better recovery rate than heat shock samples (1.11%).However, the susceptibility to Carbapenems and aminoglycosides by most isolates of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003ein this study was at variance with what was observed in other studies (Rodriguez-Palacios \u003cem\u003eet al\u003c/em\u003e., 2007; Simango and Mwakurudz, 2008; Jöbstl\u003cem\u003eet al\u003c/em\u003e., 2010) which separately reported some levels of resistance to the antibiotics by the bacterium of interest.It is noteworthy that some of these antibiotics, as mentioned above, are commonly used for the treatment of diarrhea in humans.\u003cem\u003eIn vitro,\u003c/em\u003eresistance in \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003eto several classes of antimicrobial agents hasbeen recognized and reported by Gerding (2004). Thus, there is the possibility of a high risk for CDI to occur in healthcare settings and the community at large. This finding corroborated the report of Freeman \u003cem\u003eet al.\u003c/em\u003e(2009), who asserted that the resistance of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003eto several antibacterial agents may not be essential for infection because the organismisrelatively susceptible to high-risk antimicrobial agents.\u003c/p\u003e\n\u003cp\u003eIt has been reported that the major pathogenic mechanism of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003eis the production of cytotoxin B and enterotoxin A, which are respectively encoded by \u003cem\u003etcdB\u003c/em\u003eand\u003cem\u003etcdA\u003c/em\u003egenes, co-located in a 19.6 kb region of the chromosome named pathogenicity loci (Paloc) along with other regulatory genes (Freeman \u003cem\u003eet al\u003c/em\u003e., 2009;Buddle and \u0026nbsp;Fagan 2023; Xiaolu \u003cem\u003eet al.,\u003c/em\u003e 2024).Toxin A causes diarrhea, and Toxin B is cytotoxic to the colonic cells. Virtually all studies where \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003ehave been successfully recovered have screened for the ability of the isolates to have had at least one of the three toxins or genes (\u003cem\u003etcdA, tcdB, cdtA/B\u003c/em\u003e) needed to fulfill the criteria for virulence/toxigenicity (Rodriguez-Palacios \u003cem\u003eet al\u003c/em\u003e., 2014). The findings in this study (69.56% and 100.00% for \u003cem\u003etcdA\u0026nbsp;\u003c/em\u003eand \u003cem\u003etcdB\u0026nbsp;\u003c/em\u003egenes respectively) are in agreement with the report of\u0026nbsp;Shokoohizadeh\u003cem\u003eet al\u003c/em\u003e. (2021), who opined that the genes for enterotoxin production \u003cem\u003etcdA\u003c/em\u003e are found in approximately 70% of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003eisolates, while the genes for cytotoxin production are found in all strains of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003eisolates. The severity of the disease caused by the bacterium is conferred by the presence of the genes (\u003cem\u003ecdtA\u003c/em\u003eand\u003cem\u003ecdtB\u003c/em\u003egenes), which code for the production of binary toxins (Rupnik, 2009; Bacci\u003cem\u003eet al\u003c/em\u003e., 2011). The pathogenicity of \u003cem\u003eC. difficile\u0026nbsp;\u003c/em\u003eis mainly due to the presence of two large protein toxins (toxin A and toxin B) and the fully described binary toxin (Chandrasekaran and Db, 2017).\u0026nbsp;Toxin A is a potent enterotoxinthat causes the accumulation of fluid in the gut, is cytotoxic to cells in tissue culture, and isperhaps lethal to experimental animals (Kuehne \u003cem\u003eet al.,\u003c/em\u003e 2010).Toxin B has been reported to be a 1,000-fold more potentcytotoxin than toxin A (but is not an enterotoxin) in original animal studies. Still,there are reports that some strains produce toxin B and not toxin A,which causes severe diarrhea in humans (Barbut\u003cem\u003eet al\u003c/em\u003e., 2007). More so, the reports of \u003cem\u003eC. difficile\u003c/em\u003e strains are toxin A–negative and toxin B–positive, whichhave been associated with human diseases described in certain epidemics around the world (Johnson \u003cem\u003eet al\u003c/em\u003e., 2001). The variation in the findings in this study could be a function of geographical differences and environmental conditions with respect to cultural methods and methods used for the analysis of the bacterium.\u003c/p\u003e\n\u003cp\u003eThe findings in this study are consistent with the reports from literature, which have it that the bacterium (\u003cem\u003eC. difficile\u003c/em\u003e)\u0026nbsp;from humans or animals are commonly resistant to macrolides (erythromycin) and tetracyclines as well as fluoroquinolones (moxifloxacin) and the lincosamides (clindamycin).The findings in this study are consistent with the reports from theliterature, which have it that the bacterium (\u003cem\u003eC. difficile\u003c/em\u003e) from humans or animals are commonly resistant to macrolides (erythromycin) and tetracyclines as well as fluoroquinolones (moxifloxacin) and the lincosamides (clindamycin) (Gerding, 2004; Simango and Mwakurudz, 2008; Jöbstl \u003cem\u003eet al\u003c/em\u003e., 2010; ). Also consistent with the finding in this study was the report by Knetsch\u003cem\u003eet al\u003c/em\u003e. (2018),\u0026nbsp;who opined that the global \u003cem\u003eC. difficile\u003c/em\u003e population contained a broad array of antibiotic-resistance genes encoding resistance to tetracycline and erythromycin.\u003c/p\u003e\n\u003cp\u003eThis study provides baseline data for the prevalence of \u003cem\u003eClostridium difficile\u0026nbsp;\u003c/em\u003ein clinical samples in Southeast Nigeria.\u003c/p\u003e\n\u003col\u003e\n \u003cli\u003eChances of severe infections for this pathogen in the studied location will be rare owing to the low prevalence and non-possession of genes for binary toxin production in most of the isolates in the study.\u003c/li\u003e\n \u003cli\u003eThe study has re-enacted the idea that the alcohol shock method for isolation of \u003cem\u003eClostridium difficile\u0026nbsp;\u003c/em\u003eis more effective than the heat-shock method.\u003c/li\u003e\n \u003cli\u003ePathogenic \u003cem\u003eClostridium difficile\u0026nbsp;\u003c/em\u003eisolates in the studied location were multi-drug resistant.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThis study evaluated the prevalence, molecular characterization, pathogenicity, and antibiotic sensitivity patterns of Clostridium difficile in South-Eastern Nigeria, revealing high sensitivity to gentamicin, meropenem, and amoxicillin-clavulanic acid, but resistance to tetracyclines and erythromycin, with significant expression of \u003cem\u003etcdB\u003c/em\u003e and \u003cem\u003etcdA\u003c/em\u003e virulence genes.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eCONFLICT OF INTEREST\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFUNDING\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo funding received for this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Approval and Consent to Participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was carriedout in line with the principles of the Declaration of Helsinki. Ethical approval for this research was obtained from \u003cstrong\u003eEthical Committee, Chuckwuemeka Odumegwu Ojukwu University Teachicng Hosptial Amaku, Awka, Anambra State, Nigeria.\u003c/strong\u003e The ethics approval number is\u003cstrong\u003e\u0026nbsp;COOUTH/CMAC/ETH.C/VOL.1/0035\u003c/strong\u003e and the approval was granted on \u003cstrong\u003e1/08/2018\u003c/strong\u003e. The committee reviewed and approved all aspects of the study, including the study design, recruitment methods, and data collection procedures, ensuring that they were in compliance with ethical standards and guidelines.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe \u003cstrong\u003econsent to Participate\u003c/strong\u003e was obtained from all individuals that participated in this research. Individuals were given detailed information regarding the research\u0026rsquo;s objectives, procedures, potential risks, and benefits. They were informed that participation was voluntary, that they could withdraw from the research at any given time without any punishment, and their data would be under confidentiality. All these were done prior to sample colections from the individuals\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publication\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis was done by informing all participants involved in this research about the nature of the data/material to be published, the purpose of its publication, and the potential accessibility of the publication to the general public. The individuals agrees that their details may be used in the manuscript titled \u0026ldquo;\u003cstrong\u003ePrevalence, Pathogenicity and Antimicrobial Susceptibility Pattern of \u003cem\u003eClostridioides Difficile\u003c/em\u003e From Clinical Samples in South Eastern Nigeria\u003c/strong\u003e\u0026rdquo; and published in \u003cstrong\u003eBMC Microbiology\u003c/strong\u003e \u003cstrong\u003eJournal\u003c/strong\u003e They gave their full consent knowing fully well that this redearch will contribute greatly to scientific knowledge, which could be made available in print and online formats. .Individuals were given full assurance that their confidentiality and privacy would be maintained to the fullest extent possible and that any identifying details will only be published with their explicit permission. They were brief \u0026nbsp;about their right to withdraw their consent at any time before the publication process is completed, without any punishment to their participation in the research.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailibiliy of data and material\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eand materials used in this research are not available for sharing due to\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eprivacy concerns protecting \u0026nbsp;individuals confidentiality and privacy, since it is involving human subjects, which might contain sensitive information. Materials are no longer available due to resource constraints or supply chain disruptions\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThanks to Dr. A.G. Ogofure and the members of staff of Department of Microbiology, University of Benin, Benin City, Nigeria \u0026nbsp;for their enabling environment for the Laboratory work. The authors wish to also acknowledge support received from TETFUND \u0026ndash;Centre of Excellecne for Biomedical, Engineering and Agricultural Translational studies UNIZIK, Awka, Nigeria.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAdegoke, A., Faleye, A., Singh, G. and Stenstr\u0026ouml;m, T. (2016). Antibiotic-Resistant Superbugs: Assessment of the Interrelationship of Occurrence in Clinical Settings and Environmental Niches. Molecules: \u003cem\u003eA Journal of Synthetic Chemistry and Natural Product Chemistry \u003c/em\u003e\u003cstrong\u003e22\u003c/strong\u003e. https://doi.org/10.3390/molecules22010029.\u003c/li\u003e\n\u003cli\u003eBacci, S., M\u0026oslash;lbak, K., Kjeldsen, M.K. and Olsen, K.E. (2011). 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Dissemination of Clostridium difficile in food and the environment: Significant sources of C. difficile community‐acquired infection?. \u003cem\u003eJournal of Applied Microbiology\u003c/em\u003e\u003cstrong\u003e122\u003c/strong\u003e. https://doi.org/10.1111/jam.13338.\u003c/li\u003e\n\u003cli\u003eWeese, J.S., Finley, R., Reid-Smith, R. R., Janecko, N. and Rousseau, J. (2010). Evaluation of \u003cem\u003eClostridium difficile \u003c/em\u003ein dogs and the household environment. \u003cem\u003eEpidemiological Infection. \u003c/em\u003e\u003cstrong\u003e138\u003c/strong\u003e:1100\u0026ndash;1104.\u003c/li\u003e\n\u003cli\u003eWilley, L.M., Sherwood, L.J. and Woolverton, L. (2008). \u003cem\u003eMicrobiology \u003c/em\u003e(7\u003csup\u003eth\u003c/sup\u003e Edition). Mc-Graw Hill, New York, 966 pp.\u003c/li\u003e\n\u003cli\u003eXiaolu, L., Yizhong, W., Rong, C., Fangfei, X., Xufei, W., Lin, Y., Yongmei, X., Dan, L. and Ting, Z. (2024). Antimicrobial Resistance of \u003cem\u003eClostridioides difficile\u003c/em\u003e in Children from a Tertiary Pediatric Hospital in Shanghai, China. \u003cem\u003eInfection and Drug Resistance\u003c/em\u003e \u003cstrong\u003e17\u003c/strong\u003e: 329\u0026ndash;339.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Plate","content":"\u003cp\u003ePlate 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"prevalence, molecular characterization, pathogenicity, Antibiogram, Clostridioides difficile, South-East","lastPublishedDoi":"10.21203/rs.3.rs-4955956/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4955956/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe prevalence, molecular characterization, pathogenicity and antibioticsensitivity pattern of \u003cem\u003eClostridioides difficile \u003c/em\u003efrom clinical samples in South-Eastern Nigeria wereevaluated in this study. Stool samples (450) were collected from patientsinsome selected public and private-owned clinics in the five South-Eastern states. Standard bacteriological methods were employed for enumeration of total anaerobes. Culture of \u003cem\u003eClostridioides difficile \u003c/em\u003eisolates were performed on \u003cem\u003eClostridium difficile \u003c/em\u003edifferential agar (supplemented with \u003cem\u003eC. difficile\u003c/em\u003e selective supplement). Biochemical tests like catalase, oxidase and reverse CAMP test were conducted before extraction of genomic bacterial DNA for suspected \u003cem\u003eC. difficile \u003c/em\u003eisolates. Purification and amplification of bacterial DNA was carried out on 2% agarose gel. Amplified bacteria DNA was sequenced and blasted on the National Centre for Biotechnology Information (NCBI) website. Antibiotics susceptibility was carried out for \u003cem\u003eC. difficile \u003c/em\u003eisolates using the Kirby-Bauer disc diffusion technique. PCR technique was employed for the detection of virulence (\u003cem\u003etcdA, tcdB, cdtA\u003c/em\u003eand\u003cem\u003ecdtB\u003c/em\u003e) and resistance genes (\u003cem\u003etetS, tetA\u003c/em\u003eand\u003cem\u003eermB\u003c/em\u003e) in \u003cem\u003eC. difficile \u003c/em\u003eisolates. Anaerobe counts for stool samples obtained showed that samples had a mean count of 5.63±0.09 log\u003csub\u003e10\u003c/sub\u003eCFU/g (Anambra state) while counts of 5.61±0.11 log\u003csub\u003e10\u003c/sub\u003eCFU/g and 5.65±0.07 log\u003csub\u003e10\u003c/sub\u003eCFU/g were obtained from Ebonyi and Enugu States. The antibacterial sensitivity revealed that the isolates were sensitive to gentamicin, meropenem and amoxicillin-clavulanic acid. The isolates were resistant to tetracyclines and erythromycin, and the multiple antibiotic resistance index of theisolates showed that the multi-drug resistant isolates had a MAR index of 0.44. All (14 \u003cem\u003eC. difficile \u003c/em\u003eisolates) (100%) of \u003cem\u003eC. difficile \u003c/em\u003eisolates were found to possess \u003cem\u003etcdB\u003c/em\u003egenes, and 69.56% expressed \u003cem\u003etcdA\u003c/em\u003egenes. One (4.34%) of the \u003cem\u003eC. difficile \u003c/em\u003eisolatespossessed \u003cem\u003ecdtA\u003c/em\u003eand \u003cem\u003ecdtB\u003c/em\u003egenes for binary toxin production.\u003c/p\u003e","manuscriptTitle":"Prevalence, Molecular Characterization, Pathogenicity and Antimicrobial Susceptibility Pattern of Clostridioides Difficile From Clinical Samples in South Eastern Nigeria.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-07 05:58:35","doi":"10.21203/rs.3.rs-4955956/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c09371d1-c2f8-4917-8e48-55acbc68a4c3","owner":[],"postedDate":"October 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-29T22:08:13+00:00","versionOfRecord":[],"versionCreatedAt":"2024-10-07 05:58:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4955956","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4955956","identity":"rs-4955956","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}