Funmilola A. Ayeni

Inhibition of uropathogens by lactic acid bacteria isolated from dairy foods and cow's intestine in western Nigeria

A total of 96 lactic acid bacteria (LAB) were isolated from African indigenous fermented products and cow’s intestines to study their inhibitory capability against multi-drug-resistant uropathogens. Escherichia coli accounted for approximately 45% of isolated uropathogens, followed by Staphylococcus spp. (20%). The Gram negative uropathogens were highly resistant to quinolones, co-trimoxazole, teicoplanin and some β-lactams, while the Staphylococcus spp. showed high resistance to aminoglycosides, β-lactams and macrolides. Twenty-four LAB isolates were selected based on their antimicrobial activity against two uropathogenic Staphylococcus aureus strains and bacteriocin production. LAB strains showing antimicrobial activity were grouped into smaller groups through amplified ribosomal DNA restriction analysis (ARDRA). Representative strains were identified as Weissella spp., Enterococcusfaecium, Lactococcus lactis and Lactobacillus brevis through sequencing of 16S rDNA. The Weissella spp. and L. brevis strains demonstrated remarkable inhibitory activity against seven strains of Gram negative uropathogens. Two strains of L. lactis produced a bacteriocin-like inhibitory substance active against Lactobacillus sakei. In this study, an unusual high rate of co-trimoxazole, quinolones and macrolides resistance among uropathogens from south west Nigeria was discovered. Based on their sensitivity to Weissella spp., there is a potential for using these LAB as a natural approach for the protection against the uropathogens assayed.

Antibacterial activities of lactic acid bacteria isolated from cow faeces against potential enteric pathogens

BACKGROUND The addition of sub therapeutic doses of antibiotics to cattle feed for growth promotion is a contributory factor to antibiotic resistance, thus an alternative to antibiotics is needed in animal feed additives. OBJECTIVE To determine the antimicrobial activity of cows intestinal Lactic acid bacteria (LAB) against enteric commensals. METHOD Escherichia coli, Klebsiella species (spp) and LAB were isolated from thirty different cow faecal samples and the LAB identified by partial sequencing of 16S rRNA. The antimicrobial activity of the LAB was determined against the test Escherichia coli and Klebsiella spp. RESULTS Five species of LAB were isolated from thirty cow faecal samples and identified as Enterococcus hirae (8), Enterococcus durans (6), Enterococcus faecium (1), Enterococcus faecalis (1) and Weissella confusa (1). Viable cells and cell free supernatant (CFS) of the LAB were able to inhibit the growth of the test organisms with the largest zone of inhibition by the viable cells being 26mm against Escherichia coli CB6 produced by Enterococcus hirae CO6A while Weissella confusa CO29M and Enterococcus hirae CO2A produced the largest zones of inhibition (26mm) against Klebsiella CB2. CONCLUSION This study shows that LAB from cow faeces possess considerable antimicrobial activity against resistant Escherichia coli from the same environment.

Identification and prevalence of tetracycline resistance in enterococci isolated from poultry in Ilishan, Ogun State, Nigeria.

Background: Tetracycline is one of the most frequently used antibiotics in Nigeria both for human and animal infections because of its cheapness and ready availability. The use of tetracycline in animal husbandry could lead to horizontal transfer of tet genes from poultry to human through the gut microbiota, especially enterococci. Therefore, this study is designed to identify different enterococcal species from poultry feces in selected farms in Ilishan, Ogun State, Nigeria, determine the prevalence of tetracycline resistance/genes and presence of IS256 in enterococcal strains. MaterialsandMethods: Enterococci strains were isolated from 100 fresh chicken fecal samples collected from seven local poultry farms in Ilishan, Ogun State, Nigeria. The strains were identified by partial sequencing of 16S rRNA genes. Antibiotic susceptibility of the isolates to vancomycin, erythromycin, tetracycline, gentamicin, amoxycillin/claulanate, and of loxacin were performed by disc diffusion method. Detection of tet, erm, and van genes and IS256 insertion element were done by polymerase chain reaction amplification. Results: Sixty enterococci spp. were identified comprising of Enterococcus faecalis 33 (55%), Enterococcus casseliflavus 21 (35%), and Enterococcus gallinarium 6 (10%). All the isolates were resistant to erythromycin (100%), followed by tetracycline (81.67%), amoxicillin/clavulanic acid (73.33%), ofloxacin (68.33%), vancomycin (65%), and gentamicin (20%). None of the enterococcal spp. harbored the van and erm genes while tet(M) was detected among 23% isolates and is distributed mostly among E. casseliflavus. IS256 elements were detected only in 33% of E. casseliflavus that were also positive for tet(M) gene. Conclusion: This study provides evidence that tetracycline resistance gene is present in the studied poultry farms in Ilishan, Ogun State, Nigeria and underscores the need for strict regulation on tetracycline usage in poultry farming in the studied location and consequently Nigeria.

Molecular characterization of clonal lineage and Staphylococcal toxin genes from S. aureus in Southern Nigeria

Background Staphylococcus aureus is a human colonizer with high potential for virulence, and the spread of the virulent strains from the colonized hosts to non-carriers in the community is on the increase. However, there are few reports on comprehensive analysis of staphylococcal enterotoxin (SE) genes with clonal lineage in S. aureus in Africa. This is essential because of diversity of cultures and habits of the people. This study analyzed spa types and enterotoxin genes in S. aureus strains previously isolated from the human nostrils, poultry and clinical samples in Southern Nigeria. Methods Forty-seven S. aureus isolates were obtained from humans nostrils (n = 13), clinical strains (n = 21) and poultry (n = 13) from previous studies in Southern Nigeria. The strains were analyzed for mecA gene, selected toxins genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq, ser, seu) and Panton-Valentine leukocidin (PVL) gene (lukS-PV/lukF-PV) by PCR. Population structures of the strains were detected by Staphylococcal protein A (spa) typing. Results Twenty different spa types were obtained with the highest percentages, 17% observed in spa type t091 from clinical, nasal and poultry samples while t069 was the most prevalent spa type in poultry. Two MRSA were only detected in human strains. The poultry strains had the highest occurrence of SE genes (18%) followed by nasal strains (15%) and clinical strains (10%). Eighty-nine percent of all tested isolates harbored at least one SE gene; seo was the most prevalent (34%) followed by seg (30%) and sea (21%), while sec, see and sej were absent in all strains. Spa type t355 was associated with lukS-PV/lukF-PV gene and complete absence of all studied SE. Sea, seq, seb, sek were associated with spa type t069; sea was associated with t127 while sep was associated with spa type t091. There were coexistences of seo/seg and sei/seg. Conclusions The higher carriage of staphylococci enterotoxin genes by the nasal and poultry S. aureus strains suggests a high potential of spread of staphylococcal food poisoning through poultry and healthy carriers in the community. This is the first report of high occurrence of staphylococcal enterotoxins genes in poultry from Nigeria.

Prevalence of extended spectrum beta lactamase and plasmid mediated quinolone resistant genes in strains of Klebsiella pneumonia, Morganella morganii, Leclercia adecarboxylata and Citrobacter freundii isolated from poultry in South Western Nigeria

A serious concern is arising on the coexistence of extended-spectrum beta-lactamase (ESBL) and plasmid mediated quinolone resistance (PMQR) producing bacteria in animal husbandry, which could be transferred to humans, especially in strains that may not be routinely screened for resistance. This study therefore tested the prevalence of ESBL and PMQR genes in selected bacteria isolated from poultry faeces. Faecal droppings of birds were collected from 11 farms in five states in South Western Nigeria. Bacteria were isolated from the samples on cefotaxime supplemented plates and identified with MALDI-TOF. The MIC was determined using VITEK system and resistance genes were detected with PCR. A total of 350 strains were isolated from different samples and selected strains were identified as 23 Klebsiella pneumonia, 12 Morganella morganii, seven Leclercia adecarboxylata and one Citrobacter freundii. All the species were resistant to gentamycin, trimethoprim/sulphamethaxole, tobramycin, piperacillin, cefotaxime and aztreonam (except Morganella morganii strains which were mostly susceptible to aztreonam). All the tested strains were susceptible to imipenem, meropenem and amikacin. All Leclercia adecarboxylata strains were resistant to ceftazidime, cefepime and fosfomycin while all Morganella morganii strains were resistant to fosfomycin, moxifloxacin and ciprofloxacin. All tested species were generally sensitive to ciprofloxacin except Morganella morganii strains which were resistant to ciprofloxacin. The resistance to ciprofloxacin, ceftazidime, cefepime, tigercylin, colistin and fosfomycin were 65%, 40%, 23%,, 7%, 33%, 48% respectively while the prevalence of SHV, TEM and CTX genes were 42%, 63%, 35% respectively. 9.3% of the isolates had the three ESBL genes, 2.33% had qnrA gene, 4.65% had qnr B gene while none had qnrS gene. The most prevalent PMQR gene is Oqxb (25.58%) while 6.98% had the qep gene. Klebsiella pneumoniae generally had both ESBL and PMQR genes. The high prevalence of extended spectrum beta-lactamase genes in the studied strains calls for caution in the use of beta lactam antibiotics in poultry feeds. This is the first report of the occurrence of extended spectrum beta-lactamase and plasmid mediated quinolone resistance genes in Morganella morganii and Leclercia adecarboxylata strains isolated from poultry faeces.

Characterization of Bacteria in Nigerian Yogurt as Promising Alternative to Antibiotics in Gastrointestinal Infections

ABSTRACT Gastrointestinal infections are endemic in Nigeria and several factors contribute to their continual survival, including bacterial resistance to commonly used antibiotics. Nigerian yogurts do not include probiotics, and limited information is available about the antimicrobial properties of the fermenters in the yogurt against gastrointestinal pathogens. Therefore, the antimicrobial potentials of bacteria in Nigeria-produced yogurts against intestinal pathogens were investigated in this study. Viable counts of lactic acid bacteria (LAB) in 15 brands of yogurt were enumerated and the bacteria identified by partial sequencing of 16S rRNA gene. Susceptibility of the gastrointestinal pathogens (Salmonella, Shigella and E. coli ) to antibiotics by disc diffusion method, to viable LAB by the agar overlay method, and to the cell-free culture supernatant (CFCS) of the LAB were investigated. Co-culture analysis of LAB and pathogens were also done. Viable counts of 1.5 × 1011 cfu/ml were observed in some yogurt samples. Two genera were identified: Lactobacillus (70.7%) and Acetobacter (29.3%). The Lactobacillus species reduced multidrug-resistant gastrointestinal pathogens by 4 to 5 log while the zones of inhibition ranged between 11 and 23. The Lactobacillus and Acetobacter strains examined displayed good activities against the multidrug-resistant tested pathogens. This is the first report of antimicrobial activities of acetic acid bacteria isolated from yogurt in Nigeria.

Antiviral potentials of Lactobacillus plantarum, Lactobacillus amylovorus, and Enterococcus hirae against selected Enterovirus

Enteroviruses have been associated with a host of clinical presentations including acute flaccid paralysis (AFP). The site of primary replication for most enteroviruses is the gastrointestinal tract (GIT) and lactic acid bacteria (LAB) may confer protection in the GIT against them. This study therefore investigates the antiviral potential of some selected lactic acid bacteria against enterovirus isolates recovered from AFP cases. The antiviral activities of Lactobacillus plantarum, Lactobacillus amylovorus, and Enterococcus hirae in broth culture, their cell-free supernatant (CFS), and bacterial cell pellets were assayed against Echovirus 7 (E7), E13, and E19 in a pre- and post-treatment approach using cytopathic effect (CPE) and cell viability (MTT) assay. The tested Lactobacillus plantarum, Lactobacillus amylovorus, and Enterococcus hirae strains have good antiviral properties against E7 and E19 but not against E13. Lactobacillus amylovorus AA099 shows the highest activity against E19. The pre-treatment approach displays better antiviral activities compared to post-treatment approach. The LAB in broth suspension have better antiviral activities than their corresponding CFS and bacterial pellet. Lactic acid bacteria used in this study have the potential as antiviral agents.

Characterization and anti-salmonella activities of lactic acid bacteria isolated from cattle faeces

BackgroundNon typhoidal salmonellosis is one of the neglected zoonoses in most African countries. The use of sub-therapeutic doses of antibiotics as animal growth promoter enhances the emergence and dissemination of antimicrobial resistance in bacteria with food animal reservoirs and may also results in antibiotics residue in animal products. One promising alternative to antibiotics in animal feed is Lactic Acid Bacteria (LAB) as probiotics. This study was carried out to determine the anti-salmonella activities and suitability of LAB isolated from cattle faeces in Nigeria as potential probiotics in cattle feed.MethodThe test Salmonella enterica spp strains and LAB were isolated from cattle faeces and identified by MALDI-TOF MS and partial sequencing of 16S rRNA genes respectively. The anti-salmonella activities of the isolated LAB in co-culture, cell-free supernatant, inhibition of growth by viable LAB cells and quantification of organic acids were determined by standard techniques. The ability of the LAB strains to withstand gastric conditions, antibiotic susceptibility and their haemolytic ability on blood agar were also determined.ResultsA total of 88 LAB belonging to 15 species were isolated and identified from cattle faeces. The most abundant species were Streptococcus infantarius (26), Enterococcus hirae (12), Lactobacillus amylovorus (10), Lactobacillus mucosae (10) and Lactobacillus ingluviei (9). Most of the LAB strains showed good anti-salmonella activities against the test Salmonella enterica spp. with 2 Lactobacillus strains; Lactobacillus amylovorus C94 and Lactobacillus salivarius C86 exhibiting remarkable anti-salmonella activities with total inhibition of Salmonella spp after 18 hours of co-incubation. The selected strains were able to survive simultaneous growth at pH 3 and 7% bile concentration and are non hemolytic.ConclusionThis study reports the vast diversity of culturable LAB in cattle faeces from Nigeria and their putative in-vitro antibacterial activity against Salmonella enterica spp isolated from cattle. Lactobacillus amylovorus C94 and Lactobacillus salivarius C86 demonstrated promising probiotic potentials in-vitro and will be further tested in-vivo in animal field trial.