Isolation and genotypic characterization of extended-spectrum beta-lactamase-producing Escherichia coli O157:H7 and Aeromonas hydrophila from selected freshwater sources in Southwest Nigeria.

The proliferation of antibiotic-resistant bacteria and antimicrobial resistance is a pressing public health challenge because of their possible transfer to humans via contact with polluted water sources. In this study, three freshwater resources were assessed for important physicochemical characteristics as well as heterotrophic and coliform bacteria and as potential reservoirs for extended-spectrum beta-lactamase (ESBL) strains. The physicochemical characteristics ranged from 7.0 to 8.3; 25 to 30 °C, 0.4 to 93 mg/L, 0.53 to 8.80 mg/L and 53 to 240 mg/L for pH, temperature, dissolved oxygen (DO), biological oxygen demand (BOD5) and total dissolved solids, respectively. The physicochemical characteristics mostly align with guidelines except for the DO and BOD5 in some instances. Seventy-six (76) Aeromonas hydrophila and 65 Escherichia coli O157: H7 isolates were identified by preliminary biochemical analysis and PCR from the three sites. Among these, A. hydrophila displayed higher frequencies of antimicrobial resistance, with all 76 (100%) isolates completely resistant to cefuroxime and cefotaxime and with MARI ≥ 0.61. The test isolates showed more than 80% resistance against five of the ten test antimicrobials, with resistance against cefixime, a cephalosporin antibiotic being the highest at 95% (134/141). The frequency of the detection of the resistance genes in the A. hydrophila isolates generally ranged between 0% (blaSHV) and 26.3% (blaCTX-M), while the frequency of detection among the E. coli O157:H7 isolates ranged between 4.6% (blaCTX-M) and 58.4% (blaTEM). Our findings indicate that the distribution of antibiotic-resistant bacteria with diverse ESBL-producing capabilities and virulence genes in freshwater sources potentially threatens public health and the environment.

Incidence of antibiotic resistance genotypes of Vibrio species recovered from selected freshwaters in Southwest Nigeria.

Vibrio species are classified as potent hazards because of their tendency to effect serious diseases like cholera and other gastrointestinal ailments in humans, as well as vibriosis in fish. A total of 144 freshwater samples were aseptically collected monthly across four rivers (Asejire, Ona, Dandaru and Erinle rivers) over a 12-month period from which Vibrio spp. were isolated using culture procedures, confirmed by means of biochemical test as well as Polymerase Chain Reaction (PCR) assay and further characterized for their phenotypic antibiotic susceptibilities and relevant antimicrobial resistant determinants by PCR. Three hundred and fifteen (58%) isolates confirmed across the sampled sites (Asejire = 75, Dandaru = 87, Eleyele = 72, Erinle = 81) showed high resistance against erythromycin-95%, Sulphamethoxazole-94%, rifampicin-92%, doxycycline-82%, tetracycline-75%, amoxicillin-45%, cephalothin-43% and varied susceptibilities to other antibiotics. The multiple antibiotic resistance indices of 97% of the Vibrio isolates were above the 0.2 threshold limit with MAR phenotype pattern E-SUL-RF-TET-DOX (0.38) found to be the most prevalent pattern among the isolates. The distributions of resistance determinant of the tested antibiotics were revealed as follows: sulII 33%, sulI 19% (sulfonamides); blaOXA 27%, ampC 39%, blapse 11% (beta-lactams); tetA 28%, tetE 20%, tet39 8%, (tetracyclines) and strA 39%. aacC2 24%, aphA1 14% (aminoglycosides). Strong positive associations were observed among tetA, sulI, tetE and sulII. This study raises concerns as these selected rivers may contribute to the environmental spread of waterborne diseases and antibiotic resistance genes. Therefore, we recommend environmental context-tailored strategies for monitoring and surveillance of resistance genes so as to safeguard the environment from becoming reservoirs of virulent and infectious Vibrio species.

Occurrence and antibiogram signatures of some Vibrio species recovered from selected rivers in South West Nigeria.

Vibrio species, widely distributed in water environments, has emerged as a prominent cause of water and food-related disease outbreaks posing significant risk to human and animal health worldwide. About 40% of presumptive isolates recovered from four selected rivers in Southwest Nigeria and, established as Vibrio species genus through polymerase chain reaction techniques., were subjected to antibiotic susceptibility testing against a panel of 18 commonly used antibiotics. The relative prevalence of key Vibrio species (V. parahaemolyticus, V. vulnificus, V. mimicus, V. harveyi, and V. cholerae) was in the order 17%, 13.3%, 4.4%, 2.2%, and 2.2% respectively. Antibiotic resistance by all Vibrio species was mostly observed against doxycycline (71-89%), erythromycin (86-100%), tetracycline (71-89%), rifampicin (86-100%), and sulfamethoxazole (87-100%), though susceptibility to meropenem (86-100%), cephalothin (60-100%), norfloxacin (93-100%), ciprofloxacin (88-100%), amikacin (64-100%), gentamicin (57-74%), and trimethoprim/sulfamethoxazole (57-81%) was equally observed in all species. Vibrio mimicus expressed highest resistance against streptomycin and chloramphenicol (64%), while V. vulnificus (52%) and V. cholerae (57%) had the highest resistance against cephalothin. High resistance against ampicillin (57%) and amoxicillin (50%) was exhibited by V. cholerae and V. mimicus respectively. Indexes of multiple antibiotic resistances (MARI) among Vibrio species ranged between 0.11 and 0.72 with the highest MAR index of 0.72 observed in one isolate of V. vulnificus. This study reveals high prevalence of Vibrio species in the selected rivers as well as elevated resistance against some first-line antibiotics, which suggests possible inappropriate antimicrobial usage around study communities. We conclude that the freshwater resources investigated are unfit for domestic, industrial, and recreational uses without treatment prior to use and are potential reservoirs of antibiotic-resistant Vibrio species in this environment.

Incidence and antimicrobial susceptibility fingerprints of Plesiomonas shigelliodes isolates in water samples collected from some freshwater resources in Southwest Nigeria.

Plesiomonas shigelloides, is an emerging and significant enteric pathogen in water having implication in both localised and gastrointestinal infections with characteristic of displaying high resistance against commonly used antibiotics. This study evaluated the prevalence of Plesiomonas shigelloides and their antibiogram fingerprints in water sample collected from four rivers in South-western Nigeria. In all, 148 presumptive Plesiomonas shigelloides isolates was recovered from the rivers out of which 66 (44.6%) were confirmed positive for the organism using polymerase chain reaction techniques. Confirmed isolates were evaluated for their antibiogram profiles against a panel of 20 antimicrobials using the disc diffusion method and further screened for relevant antibiotic resistance genes. Resistance of the isolates against the antimicrobials followed the order: sulphamethoxazole (100%), erythromycin (93%), ampicillin (90%), cephalotin (82%), streptomycin (64%), and chloramphenicol (58%), amoxicillin (53%), cefotaxime (50%), tetracycline (49%), neomycin (38%) and trimethoprim + sulphamethoxazole (38%). Conversely, all the isolates were susceptible against netilmicin, and susceptibility against the other antibiotics follows the order: meropenem (94%), gentamicin (88%), imipenem (79%), amikacin (70%), ciprofloxacin (70%), norfloxacin (59%), trimethoprim (56%) and ceftazidine (56%). The multiple antibiotic resistance indices of the organism were higher than the accepted threshold of 0.2. The incidence of 11 antimicrobial resistance determinants were obtained as follows: [sulphonamides; (sulI (18%), sulII (20%), dfr1 (70%), dfr(18) (5%)), [beta-lactams; (ampC 37%)], [tetracyclines; (tetA (78%), tetE (57%)], [phenicols; (catII (16%), cmlA1 (11%)] and [aminoglycosides; (aphA2 (36%) and strA (67%)]. Pearson chi-square exact test revealed positive associations among tetA, tetE, sullI and catII and tetA genes. To the best of our knowledge, this is the first report on the incidence and antibiogram fingerprint of P. shigelloides in these freshwater resources and we conclude that these rivers are important reservoirs of multiple antimicrobial resistant biotypes of this organism, and consequently a threat to public health.

Antibacterial activity of crude extracts of some South African medicinal plants against multidrug resistant etiological agents of diarrhoea.

BACKGROUND: This study evaluated the antibacterial activity of some plants used in folklore medicine to treat diarrhoea in the Eastern Cape Province, South Africa. METHODS: The acetone extracts of Acacia mearnsii De Wild., Aloe arborescens Mill., A. striata Haw., Cyathula uncinulata (Schrad.) Schinz, Eucomis autumnalis (Mill.) Chitt., E. comosa (Houtt.) Wehrh., Hermbstaedtia odorata (Burch. ex Moq.) T.Cooke, Hydnora africana Thunb, Hypoxis latifolia Wight, Pelargonium sidoides DC, Psidium guajava L and Schizocarphus nervosus (Burch.) van der Merwe were screened against Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, multi-resistant Salmonella enterica serovar Isangi, S. typhi, S. enterica serovar Typhimurium, Shigella flexneri type 1b and Sh. sonnei phase II. A qualitative phytochemical screening of the plants extracts was by thin layer chromatography. Plants extracts were screened for antibacterial activity using serial dilution microplate technique and bioautography. RESULTS: The TLC fingerprint indicated the presence of terpenoids and flavonoids in the herbs. Most of the tested organisms were sensitive to the crude acetone extracts with minimum inhibitory concentration (MIC) values ranging from 0.018-2.5 mg/mℓ. Extracts of A. striata, C. uncinulata, E. autumnalis and P. guajava were more active against enteropathogens. S. aureus and Sh. flexneri were the most sensitive isolates to the crude extracts but of significance is the antibacterial activity of A. arborescens and P. guajava against a confirmed extended spectrum betalactamase positive S. enterica serovar Typhimurium. CONCLUSION: The presence of bioactive compounds and the antibacterial activity of some of the selected herbs against multidrug resistant enteric agents corroborate assertions by traditional healers on their efficacies.

Molecular basis of virulence in clinical isolates of Escherichia coli and Salmonella species from a tertiary hospital in the Eastern Cape, South Africa.

BACKGROUND: Apart from localized gastrointestinal infections, Escherichia coli and Salmonella species are major causes of systemic disease in both humans and animals. Salmonella spp. cause invasive infections such as enteric fever, septicemia, osteomyelitis and meningitis while certain types of E. coli can cause systemic infections, includingpyelonephritis, meningitis and septicemia. These characteristic requires the involvement of a myriad of virulence factors. METHODS: This study investigated the virulence factors of Escherichia coli and Salmonella species in clinical specimens from patients with diarrhoea presenting to health care centres in Oliver R. Tambo District Municipality, Eastern Cape Province, Republic of South Africa. Microbiology analysis involved the use of cultural and molecular techniques. RESULTS: Out of a total of 315 samples screened, Salmonella isolates were obtained in 119 (37.8%) of cases and these comprised: S. choleraesuis (6%), S. enteritidis (4%), S. eppendorf (1%), S. hadar (1%), S. isangi (8%), S. panama (1%), S. typhi (52%), S. typhimurium (25%) and untyped Salmonella spp. (2%). Among the Salmonella species 87 (73.1%) were invasive. Using molecular diagnostic methods, diarrheagenic E. coli were detected in 90 cases (28.6%): the greater proportion of this were enteroaggregative E. coli (EAEC) 37 (41.1%), enteropathogenic E. coli (EPEC) 21 (23.3%) and enterohemorrhagic E. coli (EHEC) 21 (23.3%). The predominant virulence gene among the diarrheagenic E. coli was EAEC heat-stable enterotoxin astA genes while the virulence genes identified in the Salmonella strains were 15 (12.6%) flic and 105 (88.2%) inv genes. The amino acid identity of the representative genes showed 95-100% similarity to corresponding blast searched sequence. CONCLUSIONS: This study showed the diversity of virulence gene expression in two major enteric pathogens. S. typhi and enteroaggregative E. coli were the predominant enteropathogens in our study area with an indication that EAEC is endemic within our study population. It was observed among other things that some diarrheagenic E. coli isolated from apparently asymptomatic subjects expressed some virulence genes at frequency as high as seen in diarrheagenic cases. This study underlines the importance of understanding the virulence composition and diversity of pathogens for enhanced clinico-epidemiological monitoring and health care delivery.

Evaluation of the antibacterial and anticancer activities of some South African medicinal plants.

BACKGROUND: Several herbs are traditionally used in the treatment of a variety of ailments particularly in the rural areas of South Africa where herbal medicine is mainly the source of health care system. Many of these herbs have not been assessed for safety or toxicity to tissue or organs of the mammalian recipients. METHODS: This study evaluated the cytotoxicity of some medicinal plants used, inter alia, in the treatment of diarrhoea, and stomach disorders. Six selected medicinal plants were assessed for their antibacterial activities against ampicillin-resistant and kanamycin-resistant strains of Escherichia coli by the broth micro-dilution methods. The cytotoxicities of methanol extracts and fractions of the six selected plants were determined using a modified tetrazolium-based colorimetric assay (3-(4, 5-dimethylthiazol)-2, 5-diphenyl tetrazolium bromide (MTT) assay). RESULTS: The average minimum inhibitory concentration (MIC) values of the plants extracts ranged from 0.027 mg/mℓ to 2.5 mg/mℓ after 24 h of incubation. Eucomis autumnalis and Cyathula uncinulata had the most significant biological activity with the least MIC values. The in vitro cytotoxicity assay on human hepatocarcinoma cell line (Huh-7) revealed that the methanol extract of E. autumnalis had the strongest cytotoxicity with IC(50) of 7.8 µg/mℓ. Ethyl acetate and butanol fractions of C. uncinulata, Hypoxis latifolia, E. autumnalis and Lantana camara had lower cytotoxic effects on the cancer cell lines tested with IC(50) values ranging from 24.8 to 44.1 µg/mℓ; while all the fractions of Aloe arborescens and A. striatula had insignificant or no cytotoxic effects after 72 h of treatment. CONCLUSIONS: Our results indicate that the methanol fraction of E. autumnalis had a profound cytotoxic effect even though it possessed very significant antibacterial activity. This puts a query on its safety and hence a call for caution in its usage, thus a product being natural is not tantamount to being entirely safe. However, the antibacterial activities and non-cytotoxic effects of A. arborescens and A. striatula validates their continuous usage in ethnomedicine.