The potential role of Listeria monocytogenes in promoting colorectal adenocarcinoma tumorigenic process.

BACKGROUND: Listeria monocytogenes is a foodborne pathogen, which can cause a severe illness, especially in people with a weakened immune system or comorbidities. The interactions between host and pathogens and between pathogens and tumor cells have been debated in recent years. However, it is still unclear how bacteria can interact with tumor cells, and if this interaction can affect tumor progression and therapy. METHODS: In this study, we evaluated the involvement of L. monocytogenes in pre-neoplastic and colorectal cancer cell proliferation and tumorigenic potential. RESULTS: Our findings showed that the interaction between heat-killed L. monocytogenes and pre-neoplastic or colorectal cancer cells led to a proliferative induction; furthermore, by using a three-dimensional cell culture model, the obtained data indicated that L. monocytogenes was able to increase the tumorigenic potential of both pre-neoplastic and colorectal cancer cells. The observed effects were then confirmed as L. monocytogenes-specific, using Listeria innocua as negative control. Lastly, data suggested the Insulin Growth Factor 1 Receptor (IGF1R) cascade as one of the possible mechanisms involved in the effects induced by L. monocytogenes in the human colorectal adenocarcinoma cell line. CONCLUSIONS: These findings, although preliminary, suggest that the presence of pathogenic bacterial cells in the tumor niches may directly induce, increase, and stimulate tumor progression.

Recent Advances in the Control of Clinically Important Biofilms.

Biofilms are complex structures formed by bacteria, fungi, or even viruses on biotic and abiotic surfaces, and they can be found in almost any part of the human body. The prevalence of biofilm-associated diseases has increased in recent years, mainly because of the frequent use of indwelling medical devices that create opportunities for clinically important bacteria and fungi to form biofilms either on the device or on the neighboring tissues. As a result of their resistance to antibiotics and host immunity factors, biofilms have been associated with the development or persistence of several clinically important diseases. The inability to completely eradicate biofilms drastically increases the burden of disease on both the patient and the healthcare system. Therefore, it is crucial to develop innovative ways to tackle the growth and development of biofilms. This review focuses on dental- and implant-associated biofilm infections, their prevalence in humans, and potential therapeutic intervention strategies, including the recent advances in pharmacology and biomedical engineering. It lists current strategies used to control the formation of clinically important biofilms, including novel antibiotics and their carriers, antiseptics and disinfectants, small molecule anti-biofilm agents, surface treatment strategies, and nanostructure functionalization, as well as multifunctional coatings particularly suitable for providing antibacterial effects to the surface of implants, to treat either dental- or implant-related bacterial infections.

Efficacy of novel phages for control of multi-drug resistant Escherichia coli O177 on artificially contaminated beef and their potential to disrupt biofilm formation.

Contaminated beef is a prominent source of foodborne pathogens such as Escherichia coli O177. Susceptibility of nine multi-drug resistant E. coli O177 strains against eight individual phages and six phage cocktails was assessed using polystyrene microplate titer plate. Further, 180 beef samples were independently inoculated with E. coli O177 cells in triplicates and treated with eight individual phages and six phage cocktails to determine their efficacy in inhibiting bacteria growth at 4 °C over a 7-day incubation period. Results revealed that all E. coli O177 strains were susceptible to the phages. A significant log reduction in viable E. coli O177 cell counts was observed on beef samples upon phage treatment over the 7-day incubation period. Two individual phages and three phage cocktails reduced E. coli cell counts to levels below the detection limit (1.0 log10 CFU/g). Log reduction of viable E. coli cell counts ranged from 2.10 to 7.81 CFU/g for individual phages and from 2.86 to 7.81 CFU/g for cocktails. Individual phages and phage cocktails inhibited E. coli O177 biofilm formation with phage cocktails showing high efficacy. Furthermore, phage cocktails showed greater efficacy in destroying pre-formed biofilm than individual phages. Based on these findings, we concluded that phage cocktails developed in this study could be used to reduce E. coli O177 contamination and extend the shelf-life of stored raw beef.

Spectroscopic and Antibacterial Properties of CuONPs from Orange, Lemon and Tangerine Peel Extracts: Potential for Combating Bacterial Resistance.

Green synthesis of nanoparticles using citrus peel extracts is known to be environmentally friendly and non-toxic when compared to chemical methods. In this study, different citrus peel extracts obtained with the solvents acetone and distilled water were used to synthesize copper oxide nanoparticles (CuONPs). The nanoparticles were characterized using cyclic voltammetry, ultraviolet-visible spectroscopy, energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). The absorption spectrum of CuONPs prepared with acetone exhibited characteristic peaks at the wavelengths between 280-293 nm, while those with distilled water had peaks at 290 nm. The acetone-synthesized CuONPs were spherical while those produced using distilled water were rod-shaped. Based on EDS, the analysis revealed a trace spectrum of CuO nanoparticles with different weight compositions that varied with the type of citrus peel and solvent used. FTIR measurements were carried out in the range of 500-4000 cm-1 for citrus peel extract mediated CuONPs. The spectra had five vibrations occurring at approximately 473, 477, 482, 607 and 616 cm-1 for all samples, which can be attributed to the vibrations of CuO, validating the formation of highly pure CuONPs.

A Chewable Cure "Kanna": Biological and Pharmaceutical Properties of Sceletium tortuosum.

Sceletium tortuosum (L.) N.E.Br. (Mesembryanthemaceae), commonly known as kanna or kougoed, is an effective indigenous medicinal plant in South Africa, specifically to the native San and Khoikhoi tribes. Today, the plant has gained strong global attraction and reputation due to its capabilities to promote a sense of well-being by relieving stress with calming effects. Historically, the plant was used by native San hunter-gatherers and Khoi people to quench their thirst, fight fatigue and for healing, social, and spiritual purposes. Various studies have revealed that extracts of the plant have numerous biological properties and isolated alkaloids of Sceletium tortuosum are currently being used as dietary supplements for medicinal purposes and food. Furthermore, current research has focused on the commercialization of the plant because of its treatment in clinical anxiety and depression, psychological and psychiatric disorders, improving mood, promoting relaxation and happiness. In addition, several studies have focused on the isolation and characterization of various beneficial bioactive compounds including alkaloids from the Sceletium tortuosum plant. Sceletium was reviewed more than a decade ago and new evidence has been published since 2008, substantiating an update on this South African botanical asset. Thus, this review provides an extensive overview of the biological and pharmaceutical properties of Sceletium tortuosum as well as the bioactive compounds with an emphasis on antimicrobial, anti-inflammatory, anti-oxidant, antidepressant, anxiolytic, and other significant biological effects. There is a need to critically evaluate the bioactivities and responsible bioactive compounds, which might assist in reinforcing and confirming the significant role of kanna in the promotion of healthy well-being in these stressful times.

Green Synthesis of Zinc Oxide Nanoparticles from Pomegranate (Punica granatum) Extracts and Characterization of Their Antibacterial Activity.

This study assessed the antimicrobial efficacy of synthesized zinc oxide nanoparticles produced using aqueous extracts of pomegranate leaves and flowers designated ZnO-NPs-PL, ZnO-NPs-PF. In the study, oxides of zinc were successfully employed to fabricate nanoparticles using extracts from leaves and flowers of pomegranate (Punica granatum). The nanoparticles obtained were characterized spectroscopically. X-ray diffractive analysis (XRD) revealed the elemental components and nature of the synthesized particles. The fabricated zinc oxide nanoparticle (ZnO-NPs) showed a crystalline structure. The morphology of the nanoparticles as shown by scanning electron microscopy (SEM) was unevenly spherical and the functional groups involved in stabilization, reduction and capping were confirmed using Fourier Transform Infra-Red (FT-IR) Spectroscopy. Confirmation of the nanoparticles by UV-Vis analysis showed absorption bands of 284 and 357 nm for pomegranate leaf and flower extract, respectively, mediated ZnO-NPs. Evaluation of the antimicrobial efficacy of the fabricated nanoparticles showed that ZnO-NPs were effective against all selected pathogenic strains, Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, Salmonella diarizonae, Salmonella typhi, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Moraxella catarrhalis, Aeromonas hydrophila and Listeria monocytogenes, used in the analysis. The effectiveness of these nanoparticles could be linked to their sizes and shapes as obtained using a transmission electron microscope (TEM) and scanning electron microscope (SEM). Our reports revealed that increasing the concentration of the nanoparticles resulted in an increase in the antibacterial activity exerted by the nanoparticles, thus suggesting that both ZnO-NPs can effectively be used as alternative antibacterial agents. Further research is required to assess their mechanisms of action and toxicity.

Genotypic characterization of Escherichia coli O157:H7 isolates from different sources in the North-West Province, South Africa, using enterobacterial repetitive intergenic consensus PCR analysis.

In many developing countries, proper hygiene is not strictly implemented when animals are slaughtered and meat products become contaminated. Contaminated meat may contain Escherichia coli (E. coli) O157:H7 that could cause diseases in humans if these food products are consumed undercooked. In the present study, a total of 94 confirmed E. coli O157:H7 isolates were subjected to the enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR) typing to generate genetic fingerprints. The ERIC fragments were resolved by electrophoresis on 2% (w/v) agarose gels. The presence, absence and intensity of band data were obtained, exported to Microsoft Excel (Microsoft Office 2003) and used to generate a data matrix. The unweighted pair group method with arithmetic mean (UPGMA) and complete linkage algorithms were used to analyze the percentage of similarity and matrix data. Relationships between the various profiles and/or lanes were expressed as dendrograms. Data from groups of related lanes were compiled and reported on cluster tables. ERIC fragments ranged from one to 15 per isolate, and their sizes varied from 0.25 to 0.771 kb. A large proportion of the isolates produced an ERIC banding pattern with three duplets ranging in sizes from 0.408 to 0.628 kb. Eight major clusters (I-VIII) were identified. Overall, the remarkable similarities (72% to 91%) between the ERIC profiles for the isolate from animal species and their corresponding food products indicated some form of contamination, which may not exclude those at the level of the abattoirs. These results reveal that ERIC PCR analysis can be reliable in comparing the genetic profiles of E. coli O157:H7 from different sources in the North-West Province of South Africa.

Detection of vanA and vanB genes in vancomycin-resistant enterococci (VRE) from groundwater using multiplex PCR analysis.

A total of 22 groundwater samples were randomly collected from three rural communities in the Mafikeng area. Bile esculin agar was used for selective isolation of enterococci. Standard preliminary tests (Gram staining, oxidase test, catalase test) and confirmatory tests (Prolex™ Streptococcal Grouping Rapid Latex Agglutination test kit) were used to determine the identities of presumptive enterococci. The antibiotic sensitivity test was performed on all positively identified enterococci; percentage resistance and multiple antibiotic resistance (MAR) phenotypes were generated. Multiplex polymerase chain reaction (PCR) was performed to detect vanA and vanB genes vancomycin-resistant enterococci (VRE). A total of 179 enterococci were positively identified and the proportion of isolates from Dibate (62.5%) was higher compared to those from Majemantsho and Motlhabeng (22.3 and 15.0, respectively). A large proportion (81.5 to 100%) of the isolates from Dibate, Motlhabeng and Majemantsho were resistant to ampicillin, vancomycin and penicillin G. Two main MAR phenotypes, PG-VA-Ap-A-OX and PG-VA-Ap-OX, were identified. Multiplex PCR analysis of 50 VRE indicated that 17 (34%) were positive for vanA and vanB genes. This highlights the need to determine the cause of vancomycin resistance in enterococci in the sampled sites and suggests that sequence analysis be used to confirm the identities of these amplicons.

Comparative genomics and proteomics analysis of phages infecting multi-drug resistant Escherichia coli O177 isolated from cattle faeces.

The increasing prevalence of antimicrobial-resistant (AMR) pathogens has become a major global health concern. To address this challenge, innovative strategies such as bacteriophage therapy must be optimised. Genomic characterisation is a crucial step in identifying suitable phage candidates for combating AMR pathogens. The aim of this study was to characterise seven phages that infect the Escherichia coli O177 strain using a whole genome sequencing. The analysis of genome sequences revealed that these phages had linear dsDNA, with genome sizes spanning from 136, 483 to 166,791 bp and GC content varying from 35.39 to 43.63%. Taxonomically, the phages were classified under three different subfamilies (Stephanstirmvirinae, Tevenvirinae, and Vequintavirinae) and three genera (Phapecoctavirus, Tequatrovirus, and Vequintavirus) within the class Caudoviricetes. In silico PhageAI analysis predicted that all the phages were virulent, with confidence levels between 96.07 and 97.26%. The phage genomes contained between 66 and 82 ORFs, which encode hypothetical and putative functional proteins. In addition, the phage genomes contained core genes associated with molecular processes such as DNA replication, transcription modulation, nucleotide metabolism, phage structure (capsid and tail), and lysis. None of the genomes carried genes associated with undesirable traits such as integrase, antimicrobial resistance, virulence, and toxins. The study revealed high genome and proteome homology among E. coli O177 phages and other known Escherichia phages. The results suggest that the seven phages are new members of the genera Phapecoctavirus, Tequatrovirus, and Vequintavirus under the subfamilies Stephanstirmvirinae, Tevenvirinae, and Vequintavirinae, respectively.

Safety Properties of Escherichia coli O157:H7 Specific Bacteriophages: Recent Advances for Food Safety.

Shiga-toxin-producing Escherichia coli (STEC) is typically detected on food products mainly due to cross-contamination with faecal matter. The serotype O157:H7 has been of major public health concern due to the severity of illness caused, prevalence, and management. In the food chain, the main methods of controlling contamination by foodborne pathogens often involve the application of antimicrobial agents, which are now becoming less efficient. There is a growing need for the development of new approaches to combat these pathogens, especially those that harbour antimicrobial resistant and virulent determinants. Strategies to also limit their presence on food contact surfaces and food matrices are needed to prevent their transmission. Recent studies have revealed that bacteriophages are useful non-antibiotic options for biocontrol of E. coli O157:H7 in both animals and humans. Phage biocontrol can significantly reduce E. coli O157:H7, thereby improving food safety. However, before being certified as potential biocontrol agents, the safety of the phage candidates must be resolved to satisfy regulatory standards, particularly regarding phage resistance, antigenic properties, and toxigenic properties. In this review, we provide a general description of the main virulence elements of E. coli O157:H7 and present detailed reports that support the proposals that phages infecting E. coli O157:H7 are potential biocontrol agents. This paper also outlines the mechanism of E. coli O157:H7 resistance to phages and the safety concerns associated with the use of phages as a biocontrol.

Data on complete genome sequence and annotation of two multidrug resistant atypical enteropathogenic Escherichia coli O177 serotype isolated from cattle faeces.

Atypical enteropathogenic E. coli belonging to the serotype O177 is a rare strain found in ruminants, especially cattle. When compared to shiga toxin producing E. coli (STEC) O157 and non-O157 STEC (O26, O45, O103, O104, O111, O121, and O145) serotypes, the antimicrobial resistance, virulence factors, and genomic structure of E. coli O177 are poorly understood. Therefore, in this article, we present the whole genome sequence data of two aEPEC E. coli O177 isolates (E. coli O177_CF-154-A and E. coli O177_CF-335-B) generated using Illumina MiSeq platform. The raw data were generated, cleaned, and assembled using Trimmomatic and SPAdes. Genome data analysis yielded 5,112,402 and 5,460,435 bp, comprising contigs 101 and 191 with GC contents of 50.7% and 50.5% for E. coli O177_CF-154-A and E. coli O177_CF-335-B, respectively. Prokaryotic Genome Annotation Pipeline (PGAP) and Rapid Annotation using Subsystem Technology (RAST) showed that the complete genome of E. coli O177_CF-154-A contained 5040 coding sequences (CDS), 5146 genes, 4896 proteins, 90 RNAs, and 78 tRNA while that of E. coli O177_CF-335-B contained 5463 CDS, 5570 genes, 5230 proteins, 92 RNAs, and 80 tRNA for. A total of 426 and 425 subsystem features with 5190 and 5662 CDS were obtained for E. coli O177_CF-154-A and E. coli O177_CF-335-B, respectively. Several genes encoding virulence and antimicrobial resistance were identified in both genomes. Complete genome sequence data of both isolates have been deposited in the National Center for Biotechnology Information (NCBI), GenBank: accession numbers, VMKH00000000 (E. coli O177_CF-154-A) and VMKG00000000 (E. coli O177_CF-335-B). This data can be used as a reference for determining the virulence and antimicrobial resistance in E. coli O177 isolates from different sample sources.

Application of Plant-Derived Nanoparticles (PDNP) in Food-Producing Animals as a Bio-Control Agent against Antimicrobial-Resistant Pathogens.

Antibiotics are regularly used in animal husbandry to treat diseases. This practice is beneficial to animals' health and helps ensure food security. However, the misuse of antibiotics, especially in food-producing animals, has resulted in the advent of antimicrobial resistance (AMR) and its dissemination among foodborne pathogens. The occurrence of AMR in bacteria pathogens that cause infections in animals and those associated with food spoilage is now considered a global health concern affecting humans, animals and the environment. The search for alternative antimicrobial agents has kindled the interest of many researchers. Among the alternatives, using plant-derived nanoparticles (PDNPs) for treating microbial dysfunctions in food-producing animals has gained significant attention. In traditional medicine, plant extracts are considered as safe, efficient and natural antibacterial agents for various animal diseases. Given the complexity of the AMR and concerns about issues at the interface of human health, animal health and the environment, it is important to emphasize the role of a One Health approach in addressing this problem. This review examines the potential of PDNPs as bio-control agents in food-producing animals, intending to provide consumers with microbiologically safe food while ensuring food safety and security, better health for animals and humans and a safe environment.

Genetic diversity and whole genome sequence analysis data of multidrug resistant atypical enteropathogenic Escherichia coli O177 strains: An assessment of food safety and public health implications.

Atypical enteropathogenic E. coli (aEPEC) strains are emerging pathogens responsible for fatal diarrhoea in humans worldwide. The purpose of this study was to investigate genetic diversity, virulence and antimicrobial resistance profiles of aEPEC O177 strains isolated from faeces of cattle reared in intensive and extensive production systems in South Africa. A total of 96 multidrug resistant (MDR) aEPEC O177 isolates were typed using enterobacterial repetitive intergenic consensus (ERIC) and random amplified polymorphism DNA (RAPD) typing. The resistome, virulome and mobilome of two aEPEC O177 isolates were investigated using WGS analysis. The ERIC typing was efficient and reproducible with a discriminatory index of 0.95. RAPD typing had poor reproducibility with satisfactory discriminatory power of 0.859. The dendrograms constructed based on ERIC and RAPD banding patterns produced 9 and 8 clusters, respectively, which indicate genetic variation among E. coli O177 isolates. WGS analysis revealed that CF-154-A and CF-335-B) isolates belonged to the O177 serotype with H7 and H21, respectively. Both isolates harboured several virulome genes such as intimin (eaeA), haemolysin (hlyA and hlyE), translocated iron receptor (tir), Type III secretion system (eprH, gspL and prgH), bssR and bssS. However, genes encoding shiga toxins were not found in either isolate. Antibiotic resistance genes such as ampC, tet, ermB, sul2, strB AcrD, aph(6)-Ic, aph(6)-Ib, aph(3″)-I, ant (3″)-1a AcrA and acrE were found in the E. coli O177 strains. Furthermore, genome annotation results indicated that both isolates carried plasmids, insertion sequences, prophages and cluster of regularly interspaced short palindromic repeats (CRISPR) type I. Based on in silico multi locus typing (MLST) analysis, the two isolates were assigned to different sequence types (CF-154-A, ST-1308 and CF-335-B, ST-58). Whole genome multi locus typing tree showed that our isolates clustered with E. coli O177:H21 (reference), suggesting the close genomic relatedness among the strains. Overall, these findings showed that cattle carry genetically diverse E. coli O177 strains, which harbour a repertoire of virulome, resistome and mobilome genes. This highlights a need for multidrug resistant E. coli O177 strain surveillance in cattle.

Combating Bovine Mastitis in the Dairy Sector in an Era of Antimicrobial Resistance: Ethno-veterinary Medicinal Option as a Viable Alternative Approach.

Bovine mastitis (BM) is the traditional infectious condition in reared cattle which may result in serious repercussions ranging from animal welfare to economic issues. Owing to the high costs associated with preventative practices and therapeutic measures, lower milk output, and early culling, bovine mastitis is accountable for most of the financial losses suffered in cattle farming. Streptococcus agalactiae, Staphylococcus aureus, Streptococcus dysgalactiae and coliform bacteria are the predominant pathogens for bovine mastitis. In addition, the occurrence of BM has been linked to lactation stage and poor management, in the latter case, the poor stabling conditions around udder hygiene. BM occurs throughout the world, with varying rates of Streptococcus agalactiae infection in different regions. Despite the modern techniques, such as the appropriate milking practices that are applied, lower levels of pathogen vulnerability may help to prevent the development of the disease, BM treatment is primarily reliant on antibiotics for both prophylactic and therapeutic purposes. Nevertheless, as a result of the proliferation of bacterial agents to withstand the antibiotic effects, these therapies have frequently proven ineffectual, resulting in persistent BM. Consequently, alternative medicines for the management of udder inflammation have been researched, notably natural compounds derived from plants. This review focuses on BM in terms of its risk factors, pathogenesis, management, the molecular identification of causative agents, as well as the application of ethno-veterinary medicine as an alternative therapy.

Plant-derived nanoparticles as alternative therapy against Diarrheal pathogens in the era of antimicrobial resistance: A review.

Diarrhea is a condition in which feces is discharged from the bowels frequently and in a liquid form. It is one of the frequent causes of morbidity and mortality in developing countries. The impact of Diarrhea is worsened by the increasing incidence of antimicrobial resistance among the causative agents, and this is now categorized as a global healthcare challenge. Antimicrobial resistance among Diarrheal pathogens also contributes to extended infection durations, and huge economic loss even in countries with advanced public health policies. The ever-increasing incidence of antimicrobial resistance including the contraindications arising from the administration of antibiotics in some Diarrheal cases highlights a crucial need for the development of novel non-antibiotic alternative agents for therapeutic and biocontrol applications. One such intervention includes the application of plant-derived nanoparticles (PDNPs) with novel antimicrobial properties. Given their small size and large surface area to volume ratio, PDNPs can attack target bacterial cell walls to generate reactive oxygen species that may simultaneously disrupt bacteria cell components such as DNA and proteins leading to cell damage or death. This potential can make it very difficult for pathogenic organisms to develop resistance against these antibacterial agents. In this review, we provide a critical overview on the antimicrobial resistance crisis among Diarrheagenic bacteria. We also discuss the evidence from the existing literature to support the potential associated with the use of PDNPs as alternative therapeutic agents for multidrug resistant and antibiotics administer contraindicated bacteria that are associated with Diarrhea.

Evidence of Virulent Multi-Drug Resistant and Biofilm-Forming Listeria Species Isolated from Various Sources in South Africa.

Listeriosis is a foodborne disease caused by Listeria monocytogenes species and is known to cause severe complications, particularly in pregnant women, young children, the elderly, and immunocompromised individuals. The aim of this study was to investigate the presence of Listeria species in food and water using both biochemical and species-specific PCR analysis. L. monocytogenes isolates were further screened for the presence of various antibiotic resistance, virulence, and biofilm-forming determinants profiles using phenotypic and genotypic assays. A total of 207 samples (composed of meat, milk, vegetables, and water) were collected and analyzed for presence of L. monocytogenes using species specific PCR analysis. Out of 267 presumptive isolates, 53 (19.85%) were confirmed as the Listeria species, and these comprised 26 L. monocytogenes, 3 L. innocua, 2 L. welshimeri, and 1 L. thailandensis. The remaining 21 Listeria species were classified as uncultured Listeria, based on 16SrRNA sequence analysis results. A large proportion (76% to 100%) of the L. monocytogenes were resistant to erythromycin (76%), clindamycin (100%), gentamicin (100%), tetracycline (100%), novobiocin (100%), oxacillin (100%), nalidixic acid (100%), and kanamycin (100%). The isolates revealed various multi-drug resistant (MDR) phenotypes, with E-DA-GM-T-NO-OX-NA-K being the most predominant MDR phenotypes observed in the L. monocytogenes isolates. The virulence genes prfA, hlyA, actA, and plcB were detected in 100%, 68%, 56%, and 20% of the isolates, respectively. In addition, L. monocytogenes isolates were capable of forming strong biofilm at 4 °C (%) after 24 to 72 h incubation periods, moderate for 8% isolates at 48 h and 20% at 72 h (p < 0.05). Moreover, at 25 °C and 37 °C, small proportions of the isolates displayed moderate (8−20%) biofilm formation after 48 and 72 h incubation periods. Biofilm formation genes flaA and luxS were detected in 72% and 56% of the isolates, respectively. These findings suggest that proper hygiene measures must be enforced along the food chain to ensure food safety.

Behaviour of Aspergillus parasiticus in aflatoxin production as influenced by storage parameters using response surface methodology approach.

Aspergillus parasiticus is a pre-harvest and postharvest pathogen that is known to produce aflatoxin; however, it is less studied compared to A. flavus. Inappropriate storage conditions are a cause of food spoilage and growth of mycotoxigenic fungi especially in low moisture foods thus constituting hazards to health. Hence, this study investigated the behaviour of A. parasiticus on aflatoxin production in inoculated wheat flour as influenced by storage conditions using the response surface methodology. Twenty experimental runs consisting of independent variables (incubation temperature (A), time (B) and (C) moisture content) and responses (aflatoxin concentrations, i.e., AFB1, AFB2, AFG1, AFG2 and AFTOT) were developed. A central composite face-centered design was used with lower and upper limits: A (25-35 °C), B (7-15 days) and C (15-25%), while the non-inoculated wheat flour served as the negative control. Aflatoxin production was determined using High Performance Liquid Chromatography (HPLC) according to standard procedures. Numerical and graphical process variables were optimized, adequate models were predicted and optimal point prediction for aflatoxin concentration was determined. AFG1 concentrations ranged from 1.10 to 360.06 µg/g, AFG2 (0.91-446.94 µg/g), AFB2 (7.95-488.77 µg/g), AFB1 (17.21-20,666.6 µg/g) and AFTOT (15.91-21,851.09 µg/g). Aflatoxin concentration increased with increase in 'B' and 'A' but decreased with prolonged increase in 'B'. AFB1 concentrations in A. parasiticus inoculated wheat flour increased at prolonged 'B' and 'A' at constant moisture (12.09%). A reduced cubic model was significantly adequate to describe the relationship between process variables and responses (AFG1 and AFG2), cubic model (AFB1 and AFTOT) and a transformed square root cubic model for AFG2 concentrations (p ≤ 0.05). 'A' influenced AFG1 production more than 'C' while 'C' and 'A' had no significant effect on AFG2 production. Process variables 'AB' influenced AFB2 concentrations more than 'C' while 'A' had a more significant effect on the AFTOT production than 'B' (p ≤ 0.05). The predicted (R2) and adjusted coefficient of regression (adj R2) were in reasonable agreement. After optimal point prediction and validation, minimum aflatoxin concentration ≤ 0 µg/g could be achieved at the predicted conditions (A = 30.42 °C, B = 10.58 days and C = 14.49%) except in AFG2 (3.33 µg/g).

An Assessment of the Viability of Lytic Phages and Their Potency against Multidrug Resistant Escherichia coli O177 Strains under Simulated Rumen Fermentation Conditions.

Preslaughter starvation and subacute ruminal acidosis in cattle are known to promote ruminal proliferation of atypical enteropathogenic Escherichia coli strains, thereby increasing the risk of meat and milk contamination. Using bacteriophages (henceforth called phages) to control these strains in the rumen is a potentially novel strategy. Therefore, this study evaluated the viability of phages and their efficacy in reducing E. coli O177 cells in a simulated ruminal fermentation system. Fourteen phage treatments were allocated to anaerobic serum bottles containing a grass hay substrate, buffered (pH 6.6-6.8) bovine rumen fluid, and E. coli O177 cells. The serum bottles were then incubated at 39 °C for 48 h. Phage titres quadratically increased with incubation time. Phage-induced reduction of E. coli O177 cell counts reached maximum values of 61.02-62.74% and 62.35-66.92% for single phages and phage cocktails, respectively. The highest E. coli O177 cell count reduction occurred in samples treated with vB_EcoM_366B (62.31%), vB_EcoM_3A1 (62.74%), vB_EcoMC3 (66.67%), vB_EcoMC4 (66.92%), and vB_EcoMC6 (66.42%) phages. In conclusion, lytic phages effectively reduced E. coli O177 cells under artificial rumen fermentation conditions, thus could be used as a biocontrol strategy in live cattle to reduce meat and milk contamination in abattoirs and milking parlours, respectively.

First Report: Diversity of Endophytic fungi Possessing Antifungal Activity Isolated from Native Kougoed (Sceletium tortuosum L.).

Forty-three (n = 43) endophytic fungi with different morphologic characteristics were from a medicinal plant Sceletium tortuosum, were utilized to investigate their antifungal effectiveness against pathogenic fungi. All fungal isolates exhibited antifungal activity against one or more pathogens in the dual culture test whereas only 33 fungal culture filtrates (77%) showed decent antifungal effect. Fusaria and Aspergillus were the dominate genus that displayed significant antifungal activity. Isolates GG02, GG09, ND15, and ND17 showed the broadest spectrum of antifungal activity. Furthermore, culture filtrate of Fusarium sp. DR08 exhibited a broad range of antifungal activity against all the pathogens. The results suggest endophytic fungi isolated from medicinal plant might be a source of novel bioactive molecules. To the best our knowledge, this is the first report on endophytic fungi isolated from native kougoed exhibiting antifungal activity against plant fungal pathogens.

Effect of Clove (Syzygium aromaticum) spice as microbial inhibitor of resistant bacteria and Organoleptic Quality of meat.

For centuries, spices have been utilized as flavourants, colourants and as preservatives in food. Moreover, spices possess various antimicrobial properties with massive health benefits for the treatment and management of ailments and diseases. The present study was focused on three (3) aspects; (1) isolation and molecular identification of bacteria from the meat; (2) to determine the antimicrobial activity of the spices against the pathogens; (3) to assess the organoleptic properties of the spiced meat. A total of twelve (n = 12) spices evaluated against forty (n = 40) spoilage food-borne pathogenic bacteria (Escherichia coli and Enterococci spp.). The spice extracts were tested using disk diffusion method to determine the inhibition abilities. The results show that clove and black seed cumin extract exhibited excellent antibacterial activity against most pathogenic bacteria. Clove displayed the highest inhibition zone of 18 mm against E. coli (EcFwS1). Clove extract was the most inhibitor followed by black cumin, whereas extracts of thyme and cinnamon showed weak antibacterial activities against the tested strains. The most sensitive strain to spice extracts was Enterococcus spp. (EnFmL1) and the most resistant strain being E. coli. (EcFmS1 and EcFpL1). Untreated meat showed that E. coli and Enterococcus spp. count was 4.4 * 105 ± 3.4 * 105 and 2.2 * 105 ± 3.6 * 104 cfu/mL respectively after 7 days while the single dose of clove showed 5.4 * 104 ± 4.4 * 102 cfu/mL of E. coli and 1.7 * 105 ± 4.1 * 104 cfu/mL of Enterococcus spp. The organoleptic characteristics such as colour, texture, odour, pH, shape of the single dose of clove on the meat was overall acceptable.