ALBERT

Description: Biofilm is a microbial association or community attached to different biotic or abiotic surfaces or environments. These surface-attached microbial communities can be found in food, medical, industrial, and natural environments. Biofilm is a critical problem in the medical sector since it is formed on medical implants within human tissue and involved in a multitude of serious chronic infections. Food and food processing surface become an ideal environment for biofilm formation where there are sufficient nutrients for microbial growth and attachment. Therefore, biofilm formation on these surfaces, especially on food processing surface becomes a challenge in food safety and human health. Microorganisms within a biofilm are encased within a matrix of extracellular polymeric substances that can act as a barrier and recalcitrant for different hostile conditions such as sanitizers, antibiotics, and other hygienic conditions. Generally, they persist and exist in food processing environments where they become a source of cross-contamination and foodborne diseases. The other critical issue with biofilm formation is their antibiotic resistance which makes medication difficult, and they use different physical, physiological, and gene-related factors to develop their resistance mechanisms. In order to mitigate their production and develop controlling methods, it is better to understand growth requirements and mechanisms. Therefore, the aim of this review article is to provide an overview of the role of bacterial biofilms in antibiotic resistance and food contamination and emphasizes ways for controlling its production.

Description: Staphylococcus aureus is a commensal bacterium in humans and animals able to adapt to multiple environments. The aim of this study was to compare the genetic diversity and virulence profiles of strains of S. aureus isolated from food (29 strains), humans (43 strains), and animals (8 strains). 80 lipase-producing strains belonging to a biobank of 360 isolates, identified phenotypically as S. aureus, were selected. Confirmation of the species was made by amplifying the spA gene and 80% (64/80) of the strains were confirmed within this species. The virulence profile of each of the isolates was determined by PCR. The seA gene coding for enterotoxin A was found in 53.1% of the strains, the saK gene, which codes for Staphylokinase, was amplified in 57.8% of the strains, and, finally, the hlB gene coding for β-Hemolysin was amplified in 17.2%. The profile of antimicrobial resistance was determined by the Kirby Bauer method showing that the strains from food presented greater resistance to erythromycin (40.7%) and ciprofloxacin (18.5%) while in strains isolated from humans were to erythromycin (48.4%) and clindamycin (21.2%). Also, in strains from animals, a high resistance to erythromycin was observed (75%). The frequency of MRSA was 12.5% due to the presence of the mec gene and resistance to cefoxitin. Of the total strains, 68.7% were typed by PCR-RFLP of the coa gene using the AluI enzyme; derived from this restriction, 17 profiles were generated. Profile 4 (490 bp, 300 bp) was the most frequent, containing a higher number of strains with a higher number of virulence factors and antimicrobial resistance, which is associated with greater adaptation to different environments. In this study, a wide genetic diversity of strains of S. aureus from different foods, humans, and animals was found. This demonstrates evolution, genetic versatility, and, therefore, the adaptation of this microorganism in different environments.

Description: Panton–Valentine leukocidin gene is produced by Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus isolates as a pore-forming toxin is largely responsible for skin and soft tissue illnesses. MRSA produces PVL toxins through lukS and lukF proteins causing tissue necrosis by damaging membrane of the defense cells. Presence of PVL toxin was tested from the 54 S. aureus clinical isolates obtained from Thika and Kiambu Level 5 Hospitals, in Kiambu County, Kenya, by Geno Type® MRSA assay (Hain Life Science, Nehren, Germany). DNA was isolated from freshly harvested bacterial cultures by spin column using Geno Type DNA isolation kit. The detection of PVL toxins was performed by amplification of genomic DNA and by reverse hybridization that identifies PVL genes using Geno Type MRSA kit. Out of 138 samples that were collected from patients in Kiambu County, 54 S. aureus isolates were obtained, of which 14 (25.9%; 95% CI = 11.9–38.9) samples had PVL toxins. The isolates that were obtained from the female patients had a higher PVL toxin prevalence of 35.7%, while the isolates collected from the male patients had a lower prevalence of 15.4% (P=0.09). The pediatrics department had the highest PVL gene prevalence compared to outpatient department and surgical units (P=0.08). However, the age groups of patients and the hospital attended by patients showed no significant difference in terms of PVL gene prevalence (P=0.26). Therefore, the patients' gender and hospital units were not significantly associated with PVL gene prevalence (P=0.08). This study shows that PVL positive isolates occur in the sampled hospitals in the county and female as well as children must be taken into consideration among patients with wound infections when isolating S. aureus.

Description: Omics sciences and new technologies to sequence full genomes provide valuable data that are revealed only after detailed bioinformatic analysis is performed. In this work, we analyzed the genomes of seven Mexican Anaplasma marginale strains and the data from a transcriptome analysis of the tick Rhipicephalus microplus. The aim of this analysis was to identify protein sequences with predicted features to be used as potential targets to control the bacteria or tick-vector transmission. We chose three amino acid sequences different to all proteins previously reported in A. marginale that have been used as potential vaccine candidates, and also, we report, for the first time, the presence of a peroxinectin protein sequence in the transcriptome of R. microplus, a protein associated with the immune response of ticks. The bioinformatics analyses revealed the presence of B-cell epitopes in all the amino acid sequences chosen, which opens the way for their likely use as single or arranged peptides to develop new strategies for the control and prevention of bovine anaplasmosis transmitted by ticks.

Description: Leishmaniases are diseases with high epidemiological relevance and wide geographical distribution. In Brazil, Leishmania (Leishmania) amazonensis is related to the tegumentary form of leishmaniasis. The treatment for those diseases is problematic as the available drugs promote adverse effects in patients. Therefore, it is important to find new therapeutic targets. In this regard, one alternative is the study of biomolecules produced by endophytic microorganisms. In this study, the total extract produced by the endophytic Paenibacillus polymyxa RNC-D was used to evaluate the leishmanicidal, nitric oxide, and cytokines production using RAW 264.7 macrophages. The results showed that, in the leishmanicidal assay with L. amazonensis, EC50 values at the periods of 24 and 48 hours were 0.624 mg/mL and 0.547 mg/mL, respectively. Furthermore, the cells treated with the extract presented approximately 25% of infected cells with an average of 3 amastigotes/cell in the periods of 24 and 48 hours. Regarding the production of cytokines in RAW 264.7 macrophages infected/treated with the extract, a significant increase in TNF-α was observed at the periods of 24 and 48 hours in the treated cells. The concentrations of IFN-γ and IL-12 showed significant increase in 48 hours. A significant decrease in IL-4 was observed in all cells treated with the extract in 24 hours. It was observed in the treated cells that the NO production by RAW 264.7 macrophages increased between 48 and 72 hours. The endophytic Paenibacillus polymyxa RNC-D extract modulates the mediators of inflammation produced by RAW 264.7 macrophages promoting L. amazonensis death. The immunomodulatory effects might be a promising target to develop new immunotherapeutic and antileishmanial drugs.

Description: Petroleum is the major energy matrix in the world whose refining generates chemical byproducts that may damage the environment. Among such waste, polycyclic aromatic hydrocarbons (PAH) are considered persistent pollutants. Sixteen of these are considered priority for remediation, and among them is benzo(a)pyrene. Amid remediation techniques, bioremediation stands out. The genus Burkholderia is amongst the microorganisms known for being capable of degrading persistent compounds; its strains are used as models to study such ability. High-throughput sequencing allows researchers to reach a wider knowledge about biodegradation by bacteria. Using transcripts and mRNA analysis, the genomic regions involved in this aptitude can be detected. To unravel these processes, we used the model B. vietnamiensis strain G4 in two experimental groups: one was exposed to benzo(a)pyrene and the other one (control) was not. Six transcriptomes were generated from each group aiming to compare gene expression and infer which genes are involved in degradation pathways. One hundred fifty-six genes were differentially expressed in the benzo(a)pyrene exposed group, from which 33% are involved in catalytic activity. Among these, the most significant genomic regions were phenylacetic acid degradation protein paaN, involved in the degradation of organic compounds to obtain energy; oxidoreductase FAD-binding subunit, related to the regulation of electrons within groups of dioxygenase enzymes with potential to cleave benzene rings; and dehydrogenase, described as accountable for phenol degradation. These data provide the basis for understanding the bioremediation of benzo(a)pyrene and the possible applications of this strain in polluted environments.

Description: Streptomyces are widely used for the production of secondary metabolites with diverse biological activities, including antibiotics. The necessity of alternative antimicrobial agents against multidrug-resistant pathogens is indispensable. However, the production of new therapeutics is delayed in recent days. Thus, the isolation of new Streptomyces species has drawn attention. Nepal—a country rich in biodiversity—has got high possibilities for the discovery of members of actinomycetes, especially in the higher altitudes. However, in vain, only a few screening research works have been reported from Nepal to date. Streptomyces species were isolated on ISP4 media, and characterization was performed according to morphological similarity and 16S rRNA sequence similarity using bioinformatic tools. Ethyl acetate extracts of Streptomyces species were prepared, and the antimicrobial activity was carried out using agar well diffusion technique. In this report, 18 Streptomyces species isolated from the soil were reported based on sequence analysis of 16S rRNA. Among them, 12 isolates have shown antibacterial activity against extended-spectrum beta-lactamase- (ESBL-) producing Escherichia coli. Here, we have also analyzed 16S rRNA in 27 Streptomyces species whose whole-genome sequence is available, which has revealed that some species have multiple copies of the 16S gene (∼1.5 kb) with significant variation in nucleotides. In contrast, some Streptomyces species shared identical DNA sequences in multiple copies of 16S rRNA. The sequencing of numerous copies of 16S rRNA is not necessary, and the molecular sequencing of this region is not sufficient for the identification of bacterial species. The Streptomyces species-derived ethyl acetate extracts from Nepalese soil demonstrate potential activity against ESBL-producing E. coli. Thus, they are potential candidates for antibiotics manufacturing in the future.

Description: In the present study, mushrooms, Pleurotus ostreatus and Pleurotus florida, were cultivated on different agricultural wastes namely coffee straw (CS), pea straw (PS), Sorghum Grain Residue (SGR), and Wheat Grain (WG) for the evaluation of antibacterial activity. Antimicrobial activity evaluation was carried out against human pathogenic microorganisms, namely, Escherichia coli, Bacillus subtilis, Streptococcus faecalis, Pseudomonas aeruginosa, and Salmonella typhi by using the disc diffusion method. Methanolic extracts of P. ostreatus cultivated on a Sorghum grain residue substrate were recorded for the highest antibacterial activity against E. coli (19.8 mm) and P. aeruginosa (16.4 mm), and methanolic extracts of P. florida cultivated on a wheat grain substrate were recorded for the highest antibacterial activity against E. coli (18.6 mm) and S. faecalis (14.8 mm). Therefore, results suggested that P. ostreatus and P. florida cultivated on the coffee straw and Sorghum grain substrate were found with the highest antimicrobial activity in comparison to other substrates. The results supported that the methanolic extracts of P. ostreatus and P. florida might indeed be potential sources of antibacterial agents.

Description: Staphylococcus spp. is most often implicated in nosocomial infections. The objective of this study is to evaluate the susceptibility to antibiotics and the biofilm formation capacity of staphylococci species isolated from surfaces and medicotechnical materials at the university hospital center of Abomey-Calavi/Sô-Ava in Benin. Samples were collected according to ISO/DIS14698-1 standard from the surfaces and medicotechnical materials by the dry swab method. The isolation of Staphylococcus strains was performed on Chapman agar, and their identification was performed using microscopic and biochemical methods. The susceptibility of Staphylococcus isolates to antibiotics was evaluated by the disc diffusion method according to EUCAST and CLSI recommendations. The biofilm formation was qualitatively assessed using microplates. Of the 128 surfaces and medicotechnical material samples analyzed, 77% were contaminated with Staphylococcus spp. Thirteen species of Staphylococcus were isolated in different proportions but the pediatric department was the most contaminated (33%) by S. aureus. Resistance to antibiotics considerably varies according to the species of Staphylococcus. However, antibiotics such as chloramphenicol and vancomycin are the most effective on S. aureus, whereas coagulase-negative staphylococci developed less resistance to gentamycin and ciprofloxacin. The biofilm test reveals that 37% of our isolated strains were biofilm formers. Although regular monitoring of hospital hygiene is crucial, the optimal use of antibiotics is a cornerstone of reducing antimicrobial resistance.

Description: Objective. This study was aimed to evaluate the antibacterial activity of the latex of three species members of Jatropha (J. curcas, J. gossypilofia Linn., and J. multifida) against methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum beta-lactamase- (ESBL-) producing Escherichia coli and ESBL-producing Klebsiella pneumonia, carbapenemase-resistant Enterobacteriaceae (CRE)-E. coli, K. pneumoniae-carbapenemase (KPC), and carbapenemase-resistant Pseudomonas aeruginosa (CRPA). Method. The antibacterial activities were calculated based on the inhibition zones using the Mueller–Hinton agar diffusion method, minimum inhibitory concentration (MIC) using Mueller–Hinton broth in a microdilution method, and minimum bactericidal concentration (MBC) using blood agar plate. Results. The latex of Jatropha showed antibacterial activities against the MRSA and CRPA. All latex of Jatropha appeared to have the antibacterial activities against MRSA and CRPA in the diffusion method (20.4–23.7 mm and 12–15 mm), MIC (0.19–6.25%, and 25%), and MBC (0.39–12.5% and 50%). Phytochemical screening of latex indicated the presence of flavonoids. Conclusions. The latex of J. curcas, J. gossypilofia Linn., and J. multifida has the potential to be developed as antibacterial agents, especially against MRSA and CRPA strain, but further in vivo research and discovery of the mode of its action are required to shed the light on the effects.

Description: Losses associated with stem end rot (SER) of avocado fruits have been reported in all avocado growing regions of the world. In Kenya, mature avocado fruits present SER symptoms during storage and marketing, but the disease causal agent(s) has not been established. This study aimed to identify the fungal pathogen(s) associated with avocado SER in Kenya and evaluate its pathogenicity. Fungal isolates were collected from symptomatic avocado fruits from randomly selected orchards and major markets within Murang'a County, a major avocado growing region in Kenya, between September 2017 and March 2018. A total of 207 and 125 fungal isolates, recovered from orchards and major markets, respectively, were identified morphologically and further confirmed by molecular techniques. The identified isolates were Lasiodiplodia theobromae (39.8%), Neofusicoccum parvum (24.4%), Nectria pseudotrichia (18.4%), Fusarium solani (7.2%), F. oxysporum (5.1%), F. equiseti (3.9%), and Geotricum candidum (1.2%). Geotricum candidum was exclusively recovered from fruits from the market. In the pathogenicity test, L. theobromae, N. parvum, and N. pseudotrichia caused the most severe SER symptoms. Consequently, they were considered to be the major pathogens of SER of avocado fruits in Kenya. To our knowledge, this is the first report of SER pathogen of avocado fruits in Kenya. Given the significant contribution of avocado fruits to household income and foreign exchange in Kenya, this information is significant to further develop management strategies of postharvest loss of avocado fruits in Kenya.

Description: People with noninsulin-dependent diabetes mellitus are prone to urinary tract infections. There is a wide gap of information in developing countries regarding the sociodemographic factors linked to UTI among diabetics and the gender disparity among the same. Sociodemographic factors differ with geographical location and many other factors, and this makes them an important aspect that can influence the social burden of UTI among diabetics. The objective of this study was to determine the association between sociodemographic factors and UTI among diabetics. The study was carried out in the Kisii Teaching and Referral Hospital in Kenya. One hundred and eighty diabetic patients were enrolled in cross-sectional study design. Clean-catch midstream urine was collected from all participants and cultured in cysteine lactose electrolyte deficient agar for bacterial isolation. Classification of a positive culture for urinary tract infection was based on more than 100,000 (≥105) colony-forming units of a single bacterial species. The data were analyzed using frequencies, chi-square (p

Description: Seed treatment with chemical pesticides is commonly used as an initial plant protection procedure against pests and diseases. However, the use of such chemicals may impair the survival and performance of beneficial microorganisms introduced via inoculants, such as the plant growth-promoting bacterium Azospirillum brasilense. We assessed the compatibility between the most common pesticide used in Brazil for the treatment of maize seeds, composed of two fungicides, and one insecticide, with the commercial strains Ab-V5 and Ab-V6 of A. brasilense, and evaluated the impacts on initial plant development. The toxicity of the pesticide to A. brasilense was confirmed, with an increase in cell mortality after only 24 hours of exposure in vitro. Seed germination and seedling growth were not affected neither by the A. brasilense nor by the pesticide. However, under greenhouse conditions, the pesticide affected root volume and dry weight and root-hair incidence, but the toxicity was alleviated by the inoculation with A. brasilense for the root volume and root-hair incidence parameters. In maize seeds inoculated with A. brasilense, the pesticide negatively affected the number of branches, root-hair incidence, and root-hair length. Therefore, new inoculant formulations with cell protectors and the development of compatible pesticides should be searched to guarantee the benefits of inoculation with plant growth-promoting bacteria.

Description: The occurrence of carbapenem-resistant (CR) strains of Acinetobacter baumannii is reported to contribute to the severity of several nosocomial infections, especially in critically ill patients in intensive care units. The present study aims to determine the antibiotic susceptibility, clonality, and genetic mechanism of carbapenem resistance in twenty-eight Acinetobacter baumannii isolates from four hospitals in Washington DC. The antibiotic susceptibility of the isolates was determined by VITEK 2 analyses, while PCR was used to examine the presence of antibiotic-resistant genes and mobile genetic elements. Trilocus multiplex-PCR was used along with pulsed-field gel electrophoresis (PFGE) for strain typing and for accessing clonal relationships among the isolates. Antimicrobial susceptibility testing indicated that 46% of the isolates were carbapenem-resistant and possessed MDR and XDR phenotypes. PFGE clustered the 28 isolates into seven clonal (C1–C7) complexes based on 〉75% similarity cut-off. Thirty-six percent of the isolates belonged to international clone II, while 29% were assigned to Group 4 by trilocus multiplex-PCR. Although the blaOXA-51-like gene was found in all the isolates, only 36% were positive for the blaOXA-23-like gene. PCR analysis also found a metallo-β-lactamase (MBL) gene (blaVIM) in 71% of the isolates. Of the 13 CR isolates, 8 were PCR positive for both blaVIM and blaOXA-23-like genes, while 5 harbored only blaVIM gene. This study revealed the emergence of VIM carbapenemase-producing A. baumannii isolates, which has not been previously reported in the United States.

Description: Background. In most African countries including Ethiopia, Neisseria gonorrhoeae infections were diagnosed clinically and its antibiotic susceptibility was rarely tested. This study aimed to determine the prevalence and antimicrobial susceptibility patterns of N. gonorrhoeae among suspected patients attending private clinics in Jimma, Ethiopia. Methods. Institution-based cross-sectional study was conducted to determine the prevalence and antimicrobial susceptibility pattern of N. gonorrhoeae isolated from urogenital specimens. Urogenital samples were collected aseptically and then transported using Amie’s transport media and processed in a microbiology laboratory following the standard protocol. Results. Of the total 315 samples examined, 31 (9.8%) were confirmed to have gonococcal infection. Of these, 30 (96.7%) were females. High proportion of culture confirmed cases (18 (12.5%)) were observed in the 20–24 age group. All of the identified organisms were susceptible to ceftriaxone and had high resistance to penicillin (80.6%) and tetracycline (54.8%). Conclusion. The prevalence of gonococcal infection is high. In the current study, participants who have no information about sexually transmitted infection were more likely to be infected by N. gonorrhoeae. According to our study, ciprofloxacin is effective against gonococcal infection.

Description: Thermophilic Campylobacter species are clinically important aetiologies of gastroenteritis in humans throughout the world. The colonization of different animal reservoirs by Campylobacter poses an important risk for humans through shedding of the pathogen in livestock waste and contamination of water sources, environment, and food. A review of published articles was conducted to obtain information on the prevalence and antimicrobial resistance (AMR) profiles of thermophilic Campylobacter species in humans and animals in sub-Saharan Africa (SSA). Electronic databases, namely, PubMed, Google Scholar, Research4life-HINARI Health, and Researchgate.net, were searched using the following search terms “thermophilic Campylobacter,” “Campylobacter jejuni,” “Campylobacter coli,” “diarrhea/diarrhoea,” “antimicrobial resistance,” “antibiotic resistance,” “humans,” “animals,” “Sub-Saharan Africa,” and “a specific country name.” Initially, a total of 614 articles were identified, and the lists of references were screened in which 22 more articles were identified. After screening, 33 articles on humans and 34 on animals and animal products were included in this review. In humans, Nigeria reported the highest prevalence (62.7%), followed by Malawi (21%) and South Africa (20.3%). For Campylobacter infections in under-five children, Kenya reported 16.4%, followed by Rwanda (15.5%) and Ethiopia (14.5%). The country-level mean prevalence in all ages and under-five children was 18.6% and 9.4%, respectively. The prevalence ranged from 1.7%–62.7% in humans and 1.2%–80% in animals. The most reported species were C. jejuni and C. coli. The AMR to commonly used antimicrobials ranged from 0–100% in both humans and animals. Poultry consumption and drinking surface water were the main risk factors for campylobacteriosis. The present review provides evidence of thermophilic Campylobacter occurrence in humans and animals and high levels of AMR in SSA, emphasizing the need for strengthening both national and regional multisectoral antimicrobial resistance standard surveillance protocols to curb both the campylobacteriosis burden and increase of antimicrobial resistance in the region.

Description: Traditional small-scale gold mining mostly use mercury to extract the gold from ores. However, mercury contamination in the environment can affect the composition and structure of the bacterial community. The purpose of this study was to determine the effect of mercury contamination on the bacterial community in the traditional gold mining waste disposal site and in the rice field. Mercury analysis was carried out using the CVAFS method. Analysis of bacterial communities and structure was carried out based on the results of metabarcoding of the V3-V4 16S rRNA regions obtained from paired-end Illumina MiSeq reads. The results showed that the sample from the mining waste disposal site had a mercury level of 230 mg/kg, while the sample from the rice field had 3.98 mg/kg. The results showed that there were differences in microbial composition and community structure in both locations. With the total reads of 57,031, the most dominant phylum was Firmicutes in the mining disposal site sample. Meanwhile, with the total reads of 33,080, the sample from rice field was dominated by Planctomycetes. The abundant classes of bacteria in the mining waste disposal site, from the highest were Bacilli, Gammaproteobacteria and Planctomycetia, while the sample from the rice field was dominated by the Planctomycetia and Acidobacteria subdivision 6. The families that dominated the sample in disposal site were Bacillaceae and Aeromonadaceae, while the sample from the rice field was dominated by Gemmataceae. The abundant genera in both locations were Bacillus and Gemmata. This study concluded that the high level of mercury in the soil reduced the richness and diversity of bacterial phyla and lower taxa. There was also a shift in the dominance of phyla and lower taxa in both locations. This study provides an understanding of the microbial community structure in the area that is highly contaminated with mercury to open insight into the potential of these bacteria for mercury bioremediation.

Description: This study reports on the isolation and identification of Methylobacterium radiotolerans MAMP 4754 from the seeds of the medicinal plant, Combretum erythrophyllum, for the purposes of investigating antimicrobial and antioxidant activities from this endophyte. The strain identity was confirmed by 16S rRNA-based phylogeny and Scanning Electron Microscopy (SEM). Ethyl acetate and chloroform (1 : 1 v/v) extracts from the endophyte were tested for antimicrobial and antioxidant activity on a total of 7 bacterial species (3 Gram-positive and 4 Gram-negative) using the standard Minimum Inhibitory Concentration (MIC) protocol and Quantitative Radical Scavenging activity using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay, respectively. The MICs were recorded at 250 μg/mL for B. subtilis ATCC 19659, B. cereus ATCC 1076, E. coli ATCC1053, and 62.5 μg/mL for K. oxytoca ATCC 13182 and M. smegmatis ATCC 21293, while an IC50 of 5.65 μg/mL was recorded with the DPPH assay. Qualitative phytochemical analysis was positive for alkaloids, flavonoids, and steroids. Gas chromatography/mass spectrometry (GC/MS) analysis revealed the presence of 9-octadecene, 2,4-dinitrophenyl acetate, and 2(5H)-furanone, which have been previously reported for the targeted activities. M. radiotolerans MAMP 4754 tested positive for antimicrobial and antioxidant activity and this is linked to the production of plant-derived secondary metabolites by this strain.

Description: Many developing countries depend on herbal mixtures as the first line and cost-effective therapy for malaria. These mixtures with such curative tendencies may also be a source of toxicity to host cells. On the other hand, these mixtures may have anticancer potential activity characterized by cytotoxicity to cancer cells. The aim of the study was to determine the cytotoxic and antioxidant effects of five different antimalarial herbal mixtures. Five antimalarial herbal mixtures commonly used in Ghana (coded as STF, SMH, SMM, SGM, and STT) were purchased and freeze-dried. The dried samples were tested on human acute T-cell leukemia (Jurkat) and breast adenocarcinoma (MCF-7) cell lines. Cytotoxicity was assessed using the tetrazolium-based colorimetric (MTT) assay while antioxidant activity was determined using DPPH free-radical scavenging assay. Among the mixtures, SMM and SGM exhibited the strongest cytotoxicity towards Jurkat cells (IC50 values 59.17 μg/ml and 49.57 μg/ml, respectively), whereas STT showed the weakest cytotoxicity (IC50 = 244.94 μg/ml). Cytotoxic effect of SMM was also strongest towards MCF-7 cells whilst the least cytotoxic sample was SGM (IC50 〉 1000 μg/ml). SMM had the highest antioxidant percentage (EC50 = 1.05 mg/ml). The increasing order of antioxidant percentage among the five herbal mixtures is SMM 〉 SMH 〉 STT 〉 STF 〉 SGM. The herbal mixtures may be potential sources of toxic agents to host cells. Therefore, further toxicity studies must be performed to safeguard health of the public. Interestingly, cytotoxicities exhibited by SMM and SGM suggest the presence of anticancer constituents in them which warrant further studies.

Description: Proteus mirabilis is the third most common bacterium that can cause complicated UTI, especially in catheterized patients. Urovirulence genes of P. mirabilis strains are poorly identified among UTI patients. The aims of the present study were to determine the prevalence of the uropathogenic P. mirabilis strains isolated from UTI patients by the detection of several P. mirabilis virulence genes and to characterize the antibiotic susceptibility profile of P. mirabilis isolates. P. mirabilis isolates were collected from urine specimens of patients suffering from UTI. Virulence genes in P. mirabilis, namely, hpmA, hpmB, rsbA, luxS, ureC1, hlyA, rpoA, atfA, atfC, mrpA, and pm1 were detected in the isolates via PCR detection method. All P. mirabilis virulence genes were detected in more than 90% of the isolates except hlyA gene, which was detected in only 23.8% of the isolates. The rate of susceptibility for ceftriaxone was 96.8%, followed by norfloxacin (82.5%), gentamicin (71.4%), ciprofloxacin (69.8%), cephalexin (52.4%), nalidixic acid (42.9%), sulfamethoxazole (39.7%), ampicillin (36.5%), and nitrofurantoin (3.2%). Significant associations () were detected between antimicrobial susceptibility of each of the following antibiotics and the presence virulence genes. Cephalexin antimicrobial susceptibility was significantly associated with the presence each of ureC1 and atfC. Sulfamethoxazole antimicrobial susceptibility was significantly associated with the presence atfA. Ceftriaxone antimicrobial susceptibility was significantly associated with the presence each of hpmA, ureC1, rpoA, atfC, mrpA, and pm1. Nitrofurantoin antimicrobial susceptibility was significantly associated with the presence each of hpmA, ureC1, rpoA, atfA, atfC, mrpA, and pm1. In conclusion, an association between the presence of urovirulence genes of P. mirabilis and increasing P. mirabilis resistance to antimicrobials has been demonstrated.

Description: An assessment of local farmers’ knowledge, attitude, and practices on postharvest maize storage and management was carried out with a view of understanding its role in maize contamination with mycotoxins and postharvest losses in Rift Valley and Lower Eastern Regions of Kenya among 165 and 149 farmers, respectively. Differences between the two regions were analyzed using the Chi-square test, Fisher exact test, and two-sample t-test. The median quantity of maize harvested by farmers in the two regions after shelling was 585 kg. A median of 20 kg of maize was put aside as a result of rotting before shelling, and there was a significant mean difference in maize set aside as a result of rotting between the two regions (107.88 kg vs. 31.96 kg; t (306.25) = 5.707, value

Description: Aquatic ecosystems in tropical forests have a high diversity of microorganisms, including fungi, which are important decomposers of submerged wood. Despite the importance of their role in decomposition, research concerning the diversity of freshwater fungi from Brazilian Amazonian environments is scarce. The aim of this work was to describe the composition and diversity of fungi present on submerged wood in two lakes of the Brazilian Amazon (State of Pará). Fragments of decaying wood (30 samples per lake) were collected from the Lakes Juá and Maicá. The wood samples were inspected for 6 months in the presence of fungal reproductive structures. Fungi observed in the wood were identified morphologically. Twenty-three taxa were identified in the Lake Juá (10 sexual and 13 asexual) and 26 taxa in the Lake Maicá (17 sexual, 9 asexual). ITS sequences were obtained for 14 taxa to aid in identification. In the Lakes Juá and Maicá, the diversity indices were H': 2.6514 and H': 2.8174, respectively. The Sørensen index of the fungal communities in the studied lakes was 0.3673. This study is the first to describe the fungal biodiversity of two important aquatic environments in Pará, Brazil.

Description: Globally, fermented beverage and condiments are made by using different conventional practices, raw materials, and microorganisms. This paper presents the available literature review on the technology and microbiology of traditional Ethiopian beverages and condiment products. Traditional fermented beverage and condiment products have essential vitamins, minerals, enzymes, and antioxidants that are all enhanced through the process of traditional fermentation practices. In Ethiopia, fermented beverage and condiment products have practiced in a long history. During the production of traditional fermented beverage and condiment products, controlled natural fermentation process with the absence of starter cultures are used to initiate it. Moreover, the preparation of many traditionally fermented beverage and condiment products is still practiced in a household art, thereby a wide variety of fermented beverages and condiments are consumed in Ethiopia. In conclusion, the review discusses the nature of the beverage and condiment preparation, poor traditional household processing, and the extent and limitation of scientific work done so far and suggests some recommendations to limit the problem in Ethiopia.

Description: Long pepper (Piper retrofractum Vahl) is a Thai medicinal herb which has been used as one of the common ingredients in variety of Thai foods. Here, we investigated antimicrobial activities of crude bioactive metabolites extracted from fruits of P. retrofractum against 10 pathogenic organisms (bacteria and yeast) causing opportunistic infections in human or animals including Bacillus subtilis ATCC6633, Staphylococcus aureus ATCC25923, Enterococcus faecalis ATCC2921, Escherichia coli ATCC25922, Klebsiella pneumonia TISTR1843, Pseudomonas aeruginosa ATCC741, Salmonella typhi (clinical isolate), Vibrio parahaemolyticus (XN98 and 5HP), and Candida albicans ATCC90020. The results of disk diffusion test showed that the extract from methanol solvent exhibited greater antibacterial activity than other solvents with inhibition zones ranging from 0.5 to 8.0 mm, respectively. Subsequently, minimal inhibition concentration (MIC) determined by the colorimetric assay confirmed that methanol extracts showed consistent results with disk diffusion method. In summary, in vitro assays suggest that methanol is the best solvent for extraction of bioactive metabolites from P. retrofractum fruits. This crude extract can inhibit the majority of human and animal pathogens. This opens up a potential use of pepper fruits in prevention of food-contaminating microorganisms.

Description: The paper shows that the phenomenological trends of both growth and decay of a microbial population in a given medium are easily reproducible with simple equations that allow gathering the experimental data (plate counts) related to different microbial species, in different mediums and even at different temperatures, in a single master plot. The guideline of the proposed approach is that microbes and surrounding medium form a system where they affect each other and that the so-called “growth curve” is just the phenomenological appearance of such interaction. The whole system (cells and medium) changes following a definite pathway described as the evolution of a “virtual” microbial population in planktonic conditions. The proposed equations come from the assumption of a duplication mechanism with a variable generation time for the growth and of an exponential-like decline with a linear increase of the rate for the decay. The intermediate phase between growth and decay is a time span during which growth and death counterbalance each other and age differences within the virtual cell population tend to level off. The proposed approach does not provide an a priori description of this phase but allows the fit of the whole evolution trend of a microbial culture whenever the experimental data are available. Deviations of such a trend concern microbes able to form spores, modify their metabolism, or express phenotypic heterogeneity, to counterbalance adverse medium conditions.

Description: Thermophilic Campylobacter species are clinically important aetiologies of gastroenteritis in humans throughout the world. The colonization of different animal reservoirs by Campylobacter poses an important risk for humans through shedding of the pathogen in livestock waste and contamination of water sources, environment, and food. A review of published articles was conducted to obtain information on the prevalence and antimicrobial resistance (AMR) profiles of thermophilic Campylobacter species in humans and animals in sub-Saharan Africa (SSA). Electronic databases, namely, PubMed, Google Scholar, Research4life-HINARI Health, and Researchgate.net, were searched using the following search terms “thermophilic Campylobacter,” “Campylobacter jejuni,” “Campylobacter coli,” “diarrhea/diarrhoea,” “antimicrobial resistance,” “antibiotic resistance,” “humans,” “animals,” “Sub-Saharan Africa,” and “a specific country name.” Initially, a total of 614 articles were identified, and the lists of references were screened in which 22 more articles were identified. After screening, 33 articles on humans and 34 on animals and animal products were included in this review. In humans, Nigeria reported the highest prevalence (62.7%), followed by Malawi (21%) and South Africa (20.3%). For Campylobacter infections in under-five children, Kenya reported 16.4%, followed by Rwanda (15.5%) and Ethiopia (14.5%). The country-level mean prevalence in all ages and under-five children was 18.6% and 9.4%, respectively. The prevalence ranged from 1.7%–62.7% in humans and 1.2%–80% in animals. The most reported species were C. jejuni and C. coli. The AMR to commonly used antimicrobials ranged from 0–100% in both humans and animals. Poultry consumption and drinking surface water were the main risk factors for campylobacteriosis. The present review provides evidence of thermophilic Campylobacter occurrence in humans and animals and high levels of AMR in SSA, emphasizing the need for strengthening both national and regional multisectoral antimicrobial resistance standard surveillance protocols to curb both the campylobacteriosis burden and increase of antimicrobial resistance in the region.

Description: Traditional small-scale gold mining mostly use mercury to extract the gold from ores. However, mercury contamination in the environment can affect the composition and structure of the bacterial community. The purpose of this study was to determine the effect of mercury contamination on the bacterial community in the traditional gold mining waste disposal site and in the rice field. Mercury analysis was carried out using the CVAFS method. Analysis of bacterial communities and structure was carried out based on the results of metabarcoding of the V3-V4 16S rRNA regions obtained from paired-end Illumina MiSeq reads. The results showed that the sample from the mining waste disposal site had a mercury level of 230 mg/kg, while the sample from the rice field had 3.98 mg/kg. The results showed that there were differences in microbial composition and community structure in both locations. With the total reads of 57,031, the most dominant phylum was Firmicutes in the mining disposal site sample. Meanwhile, with the total reads of 33,080, the sample from rice field was dominated by Planctomycetes. The abundant classes of bacteria in the mining waste disposal site, from the highest were Bacilli, Gammaproteobacteria and Planctomycetia, while the sample from the rice field was dominated by the Planctomycetia and Acidobacteria subdivision 6. The families that dominated the sample in disposal site were Bacillaceae and Aeromonadaceae, while the sample from the rice field was dominated by Gemmataceae. The abundant genera in both locations were Bacillus and Gemmata. This study concluded that the high level of mercury in the soil reduced the richness and diversity of bacterial phyla and lower taxa. There was also a shift in the dominance of phyla and lower taxa in both locations. This study provides an understanding of the microbial community structure in the area that is highly contaminated with mercury to open insight into the potential of these bacteria for mercury bioremediation.

Description: Uganda is an agrarian country where farming employs more than 60% of the population. Aflatoxins remain a scourge in the country, unprecedentedly reducing the nutritional and economic value of agricultural foods. This review was sought to synthetize the country’s major findings in relation to the mycotoxins’ etiology, epidemiology, detection, quantification, exposure assessment, control, and reduction in different matrices. Electronic results indicate that aflatoxins in Uganda are produced by Aspergillus flavus and A. parasiticus and have been reported in maize, sorghum, sesame, beans, sunflower, millet, peanuts, and cassava. The causes and proliferation of aflatoxigenic contamination of Ugandan foods have been largely due to poor pre-, peri-, and postharvest activities, poor government legislation, lack of awareness, and low levels of education among farmers, entrepreneurs, and consumers on this plague. Little diet diversity has exacerbated the risk of exposure to aflatoxins in Uganda because most of the staple foods are aflatoxin-prone. On the detection and control, these are still marginal, though some devoted scholars have devised and validated a sensitive portable device for on-site aflatoxin detection in maize and shown that starter cultures used for making some cereal-based beverages have the potential to bind aflatoxins. More efforts should be geared towards awareness creation and vaccination against hepatitis B and hepatitis A to reduce the risk of development of liver cancer among the populace.

Description: This study is aimed at determining antibacterial activity from ethanol extracts and the most active fraction of cassava leaves against clinical isolates of Staphylococcus epidermidis and Propionibacterium acnes. Research carried out by the experimental method involved determination of plants, extraction with maceration method, fractionation with liquid-liquid extraction, antibacterial activity testing of extracts and fractions by agar diffusion method, determination of most active fraction from the extract, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) testing of most active fraction by microdilution method. The results showed that ethanol extracts of cassava leaves had antibacterial activity against both bacteria with the most active fraction indicated by ethyl acetate. MIC values of ethyl acetate fraction against S. epidermidis were in the concentration range of 2.5%–5.0% (w/v) and against P. acnes were in the concentration range of 1.25%–2.5% (w/v). The MBC value of ethyl acetate fraction against S. epidermidis was at a concentration of 5% (w/v), while P. acnes was at a concentration of 2.5% (w/v). From the results of this study, it can be concluded that the ethanol extract of cassava leaves (Manihot esculenta Crantz) has antibacterial activity against clinical isolates of Staphylococcus epidermidis as well as on Propionibacterium acnes. The fraction with the best activity from the ethanol extract of cassava leaves to the two test bacteria was shown by ethyl acetate fraction. It is suggested that cassava leaves are possible to be developed into standardized antiacne herbal.

Description: A cross-sectional study was conducted between December, 2013, and May, 2014, to determine the prevalence and antibiotic resistance feature of Salmonella isolated from broilers slaughtered in Debre Zeit and Modjo towns, Ethiopia. A total of 384 caecal content samples were collected for microbiological examination following the standard techniques and procedures outlined by the International Organization for Standardization to isolate Salmonella. The sensitivity of the isolates subjected to nine antimicrobials was tested by the Kirby–Bauer disk diffusion method. The overall prevalence of Salmonella was 14.6%, and its occurrence differ significantly by farm (). The occurrence of the bacteria was not statistically different in the midland (15.2%) and lowland (13.3%) () and between males (13.5%) and females (15.6) (). Of the 50 isolates, 48 were resistant to at least one drug. Multidrug resistance was recorded in 43 (86.0%) of the isolates. The study demonstrated considerable prevalence and high antimicrobial resistant Salmonella in exotic chicken and indicates the potential importance of chickens as source of foodborne salmonellosis and multiple antimicrobial resistance of Salmonella. Improving the hygienic practice of farms could help to reduce the occurrence of Salmonella in farms. Further studies are needed to describe the risk factors associated with the emergence of drug-resistant Salmonella in chicken.

Description: Uganda is an agrarian country where farming employs more than 60% of the population. Aflatoxins remain a scourge in the country, unprecedentedly reducing the nutritional and economic value of agricultural foods. This review was sought to synthetize the country’s major findings in relation to the mycotoxins’ etiology, epidemiology, detection, quantification, exposure assessment, control, and reduction in different matrices. Electronic results indicate that aflatoxins in Uganda are produced by Aspergillus flavus and A. parasiticus and have been reported in maize, sorghum, sesame, beans, sunflower, millet, peanuts, and cassava. The causes and proliferation of aflatoxigenic contamination of Ugandan foods have been largely due to poor pre-, peri-, and postharvest activities, poor government legislation, lack of awareness, and low levels of education among farmers, entrepreneurs, and consumers on this plague. Little diet diversity has exacerbated the risk of exposure to aflatoxins in Uganda because most of the staple foods are aflatoxin-prone. On the detection and control, these are still marginal, though some devoted scholars have devised and validated a sensitive portable device for on-site aflatoxin detection in maize and shown that starter cultures used for making some cereal-based beverages have the potential to bind aflatoxins. More efforts should be geared towards awareness creation and vaccination against hepatitis B and hepatitis A to reduce the risk of development of liver cancer among the populace.

Description: Background. Found as a commensal in the upper respiratory tract, Gram-negative diplococcus Moraxella catarrhalis did not hold much importance as an infectious agent for long. The emergence of the first antibiotic-resistant strain of M. catarrhalis was noted in 1977 in Sweden. This has gradually spread worldwide over the years to more than 95% of the strains showing resistance to penicillin now. Penicillin resistance is mediated by the production of beta-lactamases encoded by bro-1 and bro-2 genes that code for beta-lactamases BRO-1 and BRO-2, respectively. The purpose of this study was to explore the trends of antibiotic resistance, the presence of bro genes, and clinical correlation of these findings with the rise in M. catarrhalis infections worldwide. Methods. Strains of M. catarrhalis were isolated from the respiratory samples submitted to the microbiology laboratory. Preliminary identification was done using standard microbiological techniques, and antibiotic sensitivity was determined by minimum inhibitory concentration assessed using the E-test. Further, the genes associated with the development of resistance to penicillin (beta-lactamase enzyme) were detected using polymerase chain reaction technique. Results. Fourteen strains of M. catarrhalis were isolated during the study period. Majority of the strains were isolated from patients between 40 and 60 years of age and from males. Seasonality was observed with most strains being isolated during the winter season. The most important predisposing factors identified were advanced age with a history of smoking and chronic obstructive pulmonary disease. The antibiotic susceptibility pattern showed resistance to most antibiotics commonly used for the treatment of respiratory tract infections. Finally, all the strains were beta-lactamase producers, confirmed by the detection of bro-1 beta-lactamase gene in them. Conclusion. The increase in antibiotic resistance and beta-lactamase production in M. catarrhalis is a cause of concern. The emerging resistance pattern emphasises the need for an appropriate antibiotic stewardship program in clinical practice. Importance should be given to the monitoring of the trends of antibiotic susceptibility and their usage to prevent the emergence of outbreaks with resistant strains and treatment failures.

Description: Pseudomonas aeruginosa is one of the main causes of nosocomial infections and is frequently associated with opportunistic infections among hospitalized patients. Kaempferol-3-O-(2′,6′-di-O-trans-p-coumaroyl)-β-D glucopyranoside (KF) is an antipseudomonal compound isolated from the leaves of the native medicinal plant Melastoma malabathricum. Herein, an RNA-seq transcriptomic approach was employed to study the effect of KF treatment on P. aeruginosa and to elucidate the molecular mechanisms underlying the response to KF at two time points (6 h and 24 h incubation). Quantitative real-time PCR (qRT-PCR) was performed for four genes (uvrD, sodM, fumC1, and rpsL) to assess the reliability of the RNA-seq results. The RNA-seq transcriptomic analysis revealed that KF increases the expression of genes involved in the electron transport chain (NADH-I), resulting in the induction of ATP synthesis. Furthermore, KF also increased the expression of genes associated with ATP-binding cassette transporters, flagella, type III secretion system proteins, and DNA replication and repair, which may further influence nutrient uptake, motility, and growth. The results also revealed that KF decreased the expression of a broad range of virulence factors associated with LPS biosynthesis, iron homeostasis, cytotoxic pigment pyocyanin production, and motility and adhesion that are representative of an acute P. aeruginosa infection profile. In addition, P. aeruginosa pathways for amino acid synthesis and membrane lipid composition were modified to adapt to KF treatment. Overall, the present research provides a detailed view of P. aeruginosa adaptation and behaviour in response to KF and highlights the possible therapeutic approach of using plants to combat P. aeruginosa infections.

Description: Evaluation of resistance to antituberculosis drugs is routinely performed with genotypic or phenotypic methods; however, discordance can be seen between these different methodologies. Our objective was to identify mutations that could explain discordant results in the evaluation of susceptibility to rifampicin and isoniazid between molecular and phenotypic methods, using whole genome sequencing (WGS). Peruvian strains showing sensitive results in the GenoType MTBDRplus v2.0 test and resistant results in the proportions in the agar-plaque test for isoniazid or rifampin were selected. Discordance was confirmed by repeating both tests, and WGS was performed, using the Next Generation Sequencing methodology. Obtained sequences were aligned “through reference” (genomic mapping) using the program BWA with the algorithm “mem”, using as a reference the genome of the M. tuberculosis H37Rv strain. Discordance was confirmed in 14 strains for rifampicin and 21 for isoniazid, with 1 strain in common for both antibiotics, for a total of 34 unique strains. The most frequent mutation in the rpoB gene in the discordant strains for rifampicin was V170F. The most frequent mutations in the discordant strains for isoniazid were katG R463L, kasA G269S, and Rv1592c I322V. Several other mutations are reported. This is the first study in Latin America addressing mutations present in strains with discordant results between genotypic and phenotypic methods to rifampicin and isoniazid. These mutations could be considered as future potential targets for genotypic tests for evaluation of susceptibility to these drugs.

Description: Context. West Nile virus (WNV) has become endemic in many states in the United States in recent years. One in 150 individuals with West Nile virus infection develops the West Nile neuroinvasive disease (WNND) and can cause permanent and sometimes fatal neurological damage. Aims. The aim of the study was to describe the presentation characteristics and epidemiology of WNND in Louisiana to improve future recognition of cases and decrease inappropriate antibiotic use. Settings and Design. It was a retrospective descriptive-analytic cohort study. A total of 23 patients with WNND were identified at one tertiary care hospital center in Northwest Louisiana from a retrospective chart review from January 1, 2012 to October 31, 2017. Results. The median age was 49 years (range: 15–75) for patients with WNND. Of 23 patients diagnosed with WNND, twelve (52%) were diagnosed with encephalitis (WNE), six (26%) were diagnosed with meningitis (WNM), and five (22%) with myelitis (WNME). The common symptoms with WNND were fever in 65%, altered mental status in 61%, headache in 52%, fatigue in 43%, gastrointestinal symptoms in 43%, rigors in 30%, imbalance in 26%, rash in 9%, and seizures in 26% of patients. Most patients presented in the late summer season. The average duration of antibiotics given was six days. The average number of days from the admission to the diagnosis of WNND was nine days (3 to 16 days). Twenty-one (91%) patients survived the infection. Conclusions. Identifying WNV infection early in its clinical course would help in decreasing inappropriate antibiotic use when patients presented with fever and meningeal symptoms. Performing WNV serology in CSF studies is critical in making the diagnosis.

Description: Kefir is a functional beverage that contains lactic and acetic acid bacteria (LAB, AAB) and yeasts. This work’s aim was to study the chemical, microbial, and functional characteristics of kefir produced from cow’s milk and soy milk. After fermentation, free amino acids were 20.92 mg 100 mL−1 and 36.20 mg 100 mL−1 for cow’s milk and soy milk kefir, respectively. Glutamic acid was majority in both, suggesting that microbial proteolysis leads to an increase in free amino acids including glutamic acid. 108–109 CFU mL−1 LAB, 106–107 CFU mL−1 AAB, and 106–107 CFU mL−1 yeasts were counted in cow’s milk kefir, whereas soy milk kefir contained greatly lower yeasts and AAB. Lactococcus lactis, Kazachstania unispora, and Saccharomyces cerevisiae were isolated as major microorganisms in both kefirs. Acetobacter orientalis only existed in cow’s milk kefir. Cow’s milk and soy milk showed ACE inhibitory activity, which significantly increased after fermentation. Both kefirs also exhibited antioxidant activity and bactericidal activity against Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus.

Description: Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. Although their mechanism of action is not clearly explained, it is known that they positively modulate the immune system, which leads to immunity potentiation. A number of studies prove that probiotics strengthen cognitive functions, reduce anxiety, and regulate the lipid metabolism in the human body. Probiotics used in humans are most often of the Lactobacillus and Bifidobacterium species. However, as more research is conducted, new species with beneficial, probiotic properties are being discovered. This paper provides a review of available information about the influence of probiotics on human health. It summarizes the current knowledge on the mechanism of action of probiotics as well as clinical trial results proving their efficacy in allergic, neurodegenerative, and cardiac diseases. This review also discusses the data concerning the safety of probiotics in clinical treatment.

Description: Many infectious diseases are still prevalent in the world’s populations since no effective treatments are available to eradicate them. The reasons may either be the antibiotic resistance towards the available therapeutic molecules or the slow rate of producing adequate therapeutic regimens to tackle the rapid growth of new infectious diseases, as well as the toxicity of current treatment regimens. Due to these reasons, there is a need to seek and develop novel therapeutic regimens to reduce the rapid scale of bacterial infections. Antimicrobial Peptides (AMPs) are components of the first line of defense for prokaryotes and eukaryotes and have a wide range of activities against Gram-negative and Gram-positive bacteria, fungi, cancer cells, and protozoa, as well as viruses. In this study, peptides which were initially identified for their HIV inhibitory activity were further screened for antibacterial activity through determination of their kinetics as well as their cytotoxicity. From the results obtained, the MICs of two AMPs (Molecule 3 and Molecule 7) were 12.5 μg/ml for K. pneumoniae (ATCC 700603) and 6.25 μg/ml for P. aeruginosa (ATCC 22108). The two AMPs killed these bacteria rapidly in vitro, preventing bacterial growth within few hours of treatment. Furthermore, the cytotoxic activity of these two peptides was significantly low, even at an AMP concentration of 100 μg/ml. These results revealed that Molecule 3 and 7 have great potential as antibacterial drugs or could serve as lead compounds in the design of therapeutic regimens for the treatment of antibiotic-resistant bacteria.

Description: Microbes are found all over the globe with some few exceptions, including sterilized surfaces. They include normal flora that is nonpathogenic, which contribute to the larger percentage, and pathogenic species which are few. Hence, the activities of humans cannot be completely separated from microbes. Thus, many pathogenic microbes have found their way into fresh fruits and vegetables which are a great source of a healthy diet for humans. The growing demand for fresh fruits and vegetables has necessitated larger production. The larger production of vegetables within the shortest possible time to meet the growing demand has placed them at a higher risk of contamination with the pathogenic microbes, making the safety of consumers uncertain. Study of sources of contamination and type of pathogenic etiological agents isolated from fresh fruits and vegetables includes Bacillus cereus, Campylobacter jejuni, Clostridium botulinum, E. coli O157: H7, Listeria monocytogenes, Salmonella spp., Shigella, Staphylococcus, and Vibrio cholera. Several measures have proven to be effective in controlling contamination of microbes and they include the establishment of surveillance systems to monitor the production chain and thoroughly washing vegetables with vinegar water. Saltwater and other washing techniques are effective but caution should be taken to make sure one does not use one cycle of water for washing all vegetables. The consumption of fresh fruits and vegetables is still encouraged by this review but significant measures must be taken to check the safety of these products before consumption.

Description: In this study, five bacteriocin-producing Lactococcus lactis strains were identified from different naturally fermented Brazilian sausages. Ion exchange and reversed-phase chromatographies were used to purify the bacteriocins from culture supernatant of the five strains. Mass spectrometry (MALDI-TOF/TOF) showed that the molecular masses of the bactericoins from L. lactis ID1.5, ID3.1, ID8.5, PD4.7, and PR3.1 were 3330.567 Da, 3330.514 Da, 3329.985 Da, 3329.561 Da, and 3329.591 Da, respectively. PCR product sequence analysis confirmed that the structural genes of bacteriocins produced by the five isolates are identical to the lantibiotic nisin Z. Optimal nisin Z production was achieved in tryptone and casein peptone, at pH 6.0 or 6.5. The most favorable temperatures for nisin Z production were 25°C and 30°C, and its production was better under aerobic than anaerobic condition. The type of carbon source appeared to be an important factor for nisin Z production. While sucrose was found to be the most efficient carbon source for nisin Z production by four L. lactis isolates, fructose was the best for one isolate. Lactose was also a good energy source for nisin Z production. Surprisingly, glucose was clearly the poorest carbon source for nisin Z production. The five isolates produced different amounts of the bacteriocin, L. lactis ID1.5 and ID8.5 isolates being the best nisin Z producers. DNA sequence analysis did not reveal any sequence differences in the nisZ and nisF promoter regions that could explain the differences in nisin Z production, suggesting that there should be other factors responsible for differential nisin Z production by the isolates.

Description: The quality of drinking water is a powerful environmental determinant of health. Water becomes contaminated with faecal material due to inadequate protection of the source, unhygienic practices of the community at the source, and poor household handling practices. The objective of this study was to assess the level of bacteriological contamination of drinking water supply from protected water sources to point of use and water handling practices among beneficiary households of Boloso Sore woreda, Wolaita zone, Ethiopia. A cross-sectional survey and bacteriological analysis of water were conducted in January 2019. The study included 545 households for water handling practices, and 75 samples from stored water from households and eighteen water sources were included for faecal coliform test. Data were analyzed using SPSS v21.0. Descriptive and logistic regression statistical models were used. Sixty percent of shallow wells, 60% of protected hand-dug wells, and 25% of protected on-spot springs were found positive for faecal coliform. In general, 44% of water source samples and 91% of household water samples were positive for faecal coliform. In general, 38% of households were practicing unsafe water handling practices. High school and above level of education (AOR = 3.37, 95% CI: 1.03, 11.57), getting higher monthly income (AOR = 2.37, 95%CI: 1.96, 5.85), households with small family size (AOR = 1.81, 95% CI: 1.15, 2.83), frequency of water collection twice a day (AOR = 2.88, 95% CI:1.56, 5.33), and presence of water payments (AOR = 0.42, 95% CI: 0.24, 0.72) were significantly associated with water handling practice. Unsafe water handling was a common practice in the study area, and water sources and household water storage were not free of faecal coliform, indicating noncompliance with the World Health Organization water quality guideline. Hence, capacity building is mandatory for the protection and management of water sources and safe water handling practices in the household and community.

Description: Salmonella is one of the most harmful pathogens responsible for foodborne outbreaks, illnesses and deaths. The aim of this study was to evaluate the effect of potentially probiotic strains against Salmonella Typhimurium DT104 in mice. The compatibility test among the selected potential probiotic strains (Lactobacillus plantarum K132, Lactobacillus paracasei K114 and Lactococcus lactis E124) using the cross-streaking method showed the absence of antagonism. The anti-Salmonella activities of coculture of the isolated potential probiotics in the form of mixed or single culture showed a remarkable anti-Salmonella activity with 96.50 to 100% growth inhibition. The combination of strains, which showed the highest growth inhibition rates against Salmonella Typhimurium DT104, was used to test their effect on the colonization of mice by Salmonella Typhimurium DT104. White albino male mice were pretreated with the mixed potential probiotics for 7 days and infected with Salmonella Typhimurium DT104 for 1 day. A total of 3 treatments were applied, during which the negative control group was treated with phosphate-buffered saline (PBS); a positive control group (typ) was challenged with Salmonella Typhimurium DT104 alone. The treated group (pro-typ) was pretreated with mixed potential probiotic culture and then infected with Salmonella Typhimurium DT104. The survival rate of mice and counts of Salmonella in feces were recorded. The survival rate of mice on day 21 after the oral challenge with Salmonella Typhimurium DT104 was significantly p

Description: Supershedding cattle shed Escherichia coli O157:H7 (O157) at ≥ 104 colony-forming units/g feces. We recently demonstrated that a supershed O157 (SS-O157) strain, SS-17, hyperadheres to the rectoanal junction (RAJ) squamous epithelial (RSE) cells which may contribute to SS-O157 persistence at this site in greater numbers, thereby increasing the fecal O157 load characterizing the supershedding phenomenon. In order to verify if this would be the signature adherence profile of any SS-O157, we tested additional SS-O157 isolates (n = 101; each from a different animal) in the RSE cell adherence assay. Similar to SS-17, all 101 SS-O157 exhibited aggregative adherence on RSE cells, with 56% attaching strongly (〉10 bacteria/cell; hyperadherent) and 44% attaching moderately (1–10 bacteria/cells). Strain typing using Polymorphic Amplified Typing Sequences (PATS) analysis assigned the 101 SS-O157 into 5 major clades but not to any predominant genotype. Interestingly, 69% of SS-O157 isolates were identical to human O157 outbreak strains based on pulsed field gel electrophoresis profiles (CDC PulseNet Database), grouped into two clades by PATS distinguishing them from remaining SS-O157, and were hyperadherent on RSE cells. A subset of SS-O157 isolates (n = 53) representing different PATS and RSE cell adherence profiles were analyzed for antibiotic resistance (AR). Several SS-O157 (30/53) showed resistance to sulfisoxazole, and one isolate was resistant to both sulfisoxazole and tetracycline. Minimum inhibitory concentration (MIC) tests confirmed some of the resistance observed using the Kirby–Bauer disk diffusion test. Each SS-O157 isolate carried at least 10 genes associated with AR. However, genes directly associated with AR were rarely amplified: aac (3)-IV in 2 isolates, sul2 in 3 isolates, and tetB in one isolate. The integrase gene, int, linked with integron-based AR acquisition/transmission, was identified in 92% of SS-O157 isolates. Our results indicate that SS-O157 isolates could potentially persist longer at the bovine RAJ but exhibit limited resistance towards clinical antibiotics.

Description: In this study, hydrodistillation was used to obtain essential oils (EOs) from pepper (Piper nigrum L.) and clove (Eugenia caryophyllata) and co-hydrodistillation (addition of fatty acid ethyl esters as extraction co-solvents) was used to obtain functional extracts (FEs). Antifungal activity of EOs and FEs was evaluated by determination of minimum inhibitory concentration (MIC) against Fusarium oxysporum and Aspergillus niger. The results showed that pepper (Piper nigrum) and clove (Eugenia caryophyllata) essential oils and their functional extracts are effective in vitro at concentrations from 400 to 500 ppm after 10 days of culturing. The essential oils and functional extracts were used on tomato fruit samples at three different concentrations: 350, 400, and 450 ppm5. Clove essential oil reduced the growth of Aspergillus niger from 50% to 70% and Fusarium oxysporum to 40%. The functional extracts (FEs) of clove and pepper, mixed with ethyl decanoate (FEs-C10), were the best combination for protecting the tomato fruit in vivo against both phytopathogenic fungi. Gas chromatography-mass spectrometry (GC-MS) was used to identify eugenol as the principal compound in clove oil and limonene, sabinene, and β-caryophyllene in pepper oil.

Description: Indonesian marine environments are known to house diverse organisms. However, the potential for bacteria from these environments as a source of antibacterial agents has not been widely studied. This study aims to explore the antibacterial potential of secondary metabolites produced by bacterial symbionts from sponges and corals collected in the Indonesian waters. Extracts of 12 bacterial isolates from sponges or corals were prepared by cultivating the bacteria under a number of different media conditions and using agar well diffusion assays to test for antibacterial activity. In addition, the morphology, physiology, and biochemical characteristics and 16S rRNA sequence of each isolate were used to determine their taxonomic classification. All tested bacterial isolates were able to produce secondary metabolites with various levels of antibacterial activity depending on medium composition and culture conditions. Two of the bacteria (RS3 and RC4) showed strong antibacterial activities against both Gram-negative and Gram-positive bacteria. A number of isolates (RS1, RS3, and RC2) were co-cultured with mycolic acid-containing bacteria, Mycobacterium smegmatis or Rhodococcus sp. However, no improvements in their antibacterial activity were observed. All of the 12 bacteria tested were identified as Streptomyces spp. LC-MS analysis of EtOAc extracts from the most active strains RS3 and RC4 revealed the presence of a number of dactinomycin analogues and potentially new secondary metabolites. Symbiotic Streptomyces spp. from sponges and corals of the Indonesian marine environments have great potential as a source of broad-spectrum antibacterial agents.

Description: Background. Food-borne disease is mostly caused by unsafe food handling and processing as well as poor hygienic practice. Recently, it is a worldwide and local burden to the human health. It is estimated that about one-third of the world population is affected by food-borne diseases annually and become a global public health problem. Hence, this study aimed to determine the prevalence, antimicrobial susceptibility patterns, and associated risk factors of Shigella, Salmonella, and intestinal parasites among food handlers in Dilla University, Southern Ethiopia. Methods. An institutional-based cross-sectional study was conducted from November to September 2018/2019. A structured questionnaire was used for the collection of data on sociodemographic characteristics. Parasite and bacterial identification, as well as susceptibility testing, was done using standard parasitological and bacteriological procedures. Results. Of the total 220 food handlers included in the study, 38.6%, 9.5%, and 3.2% were positive for intestinal parasites, Salmonella, and Shigella species, respectively. A. lumbricoides (11.4%) was the predominant parasite isolated followed by E. histolytica (7.7%). From the total Salmonella isolates, serogroup D was the most frequent isolate and from the total Shigella species, Shigella flexneri was the predominant isolate. In this study, through irregular medical checkups, those who drunk unpasteurized milk and ate raw meat were significantly associated with intestinal parasites. Both Salmonella and Shigella species were highly resistant to ampicillin (81%) and amoxicillin-clavulanic acid (〉70%). Salmonella isolates are highly sensitive to cefotaxime and ceftriaxone, while Shigella is highly sensitive to ciprofloxacin and norfloxacin. MDR was recorded in 71.4% of all bacterial isolates. Conclusion. The presence of a high prevalence of intestinal parasites, Salmonella, and Shigella species that were resistant to the commonly prescribed drugs is a treat to the children and the community at large. Therefore, measures including health education, improvement of safe water supply, sanitation facilities, and continuous monitoring of microbiological and antimicrobial surveillance are crucial.

Description: Background. Urinary tract infections are the common types of infections in the community and health care settings. Despite the widespread availability of antibiotics, urinary tract infection remains a worldwide therapeutic problem. It is a continuous and significant problem in cancer patients. Methods. A hospital-based comparative cross-sectional study was conducted on 240 study participants from January to June 2019. Sociodemographic data were collected by a predesigned questionnaire and midstream urine samples collected using simple random sampling technique by using clean, sterile plastic cups and then inoculated onto CLED agar plates and incubated at 37°C for 24 hours. Urine culture was considered significant bacteriuria when colony forming units ≥105/mL of voided urine and a single pure colony suspended in nutrient broth and then subcultured onto a blood agar plate and MacConkey agar plate, incubated at 37°C for 24 hours for identification. Identification was done by using standard microbiological methods. Modified Kirby–Bauer disk diffusion technique was applied for antimicrobial susceptibility testing in accordance with CLSI 2018 criteria. Data were entered, cleared, and checked using Epi Info version 7 and exported to SPSS version 20 for analysis. The results were displayed using tables and figures. p value

Description: Aspergillus flavus is one of the most common isolates from patients with fungal infections. Aspergillus infection is usually treated with antifungal agents, but side effects of these agents are common. Trehalase is an essential enzyme involved in fungal metabolism, and the trehalase inhibitor, validamycin A, has been used to prevent fungal infections in agricultural products. In this study, we observed that validamycin A significantly increased trehalose levels in A. flavus conidia and delayed germination, including decreased fungal adherence. In addition, validamycin A and amphotericin B showed a combinatorial effect on A. flavus ATCC204304 and clinical isolates with high minimum inhibitory concentrations (MICs) of amphotericin B using checkerboard assays. We observed that validamycin A and amphotericin B had a synergistic effect on A. flavus strains resistant to amphotericin B. The MICs in the combination of validamycin A and amphotericin B were at 0.125 μg/mL and 2 μg/mL, respectively. The FICI of validamycin A and amphotericin B of these clinical isolates was about 0.25–0.28 with synergistic effects. No drug cytotoxicity was observed in human bronchial epithelial cells treated with validamycin A using LDH-cytotoxicity assays. In conclusion, this study demonstrated that validamycin A inhibited the growth of A. flavus and delayed conidial germination. Furthermore, the combined effect of validamycin A with amphotericin B increased A. flavus killing, without significant cytotoxicity to human bronchial epithelial cells. We propose that validamycin A could potentially be used in vivo as an alternative treatment for A. flavus infections.

Description: Introduction. Carbapenem-resistant Acinetobacter baumannii has been responsible for an increasing number of hospital-acquired infections globally. The study investigated the prevalence of carbapenemase-encoding genes in clinical multidrug-resistant A. baumannii strains. Materials and Methods. A total of 100 nonduplicate multidrug-resistant A. baumannii strains were cultured from clinical samples obtained from healthcare facilities in the O. R. Tambo district. The strains were confirmed by detecting the intrinsic blaOXA-51-like gene. Antimicrobial susceptibility testing was performed by VITEK® 2 and autoSCAN-4 systems. The MIC of imipenem and meropenem was rechecked by E-test. Colistin MIC was confirmed by the broth microdilution method. Real-time PCR was performed to investigate the presence of carbapenemase-encoding genes. Results. Most strains showed high resistance rates (〉80%) to the antibiotics tested. Resistance to amikacin, tetracycline, and tigecycline were 50%, 64%, and 48%, respectively. All strains were fully susceptible to colistin. The blaOXA-51-like was detected in all strains whilst blaOXA-23-like, blaOXA-58-like, blaOXA-24-like, blaIMP-1, blaVIM, and blaNDM-1 were found in 70%, 8%, 5%, 4%, 3%, and 2% of strains, respectively. None of the tested strains harboured the genes blaSIM and blaAmpC. The coexistence of blaOXA-23-like, and blaIMP-1 or blaOXA-58-like was detected in 1% and 2% strains, respectively. A distinct feature of our findings was the coharbouring of the genes blaOXA-23-like, blaOXA-58-like, and blaIMP-1 in 2% strains, and this is the first report in the Eastern Cape Province, South Africa. The intI1 was carried in 80% of tested strains whilst ISAba1/blaOXA-51-like and ISAba1/blaOXA-23-like were detected in 15% and 40% of the strains, respectively. The detection of blaOXA-23-like, ISAba1/blaOXA-51-like, ISAba1/blaOXA-23-like, and blaOXA-23-like, blaOXA-58-like, and blaIMP-1 carbapenemases in strains had a significant effect on both imipenem and meropenem MICs. Conclusions. Results showed a high level of oxacillinases producing A. baumannii circulating in our study setting, highlighting the need for local molecular surveillance to inform appropriate management and prevention strategies.

Description: Cassava (Manihot esculenta Crantz) is a major source of carbohydrates, calcium, vitamins (B and C), and essential minerals and is the third most important source of calories in the tropics. However, it is not clear if the traditional processing methods expose the products to microbial contamination. This study assessed the levels of fungi and aflatoxin contamination in traditionally processed cassava products (Akuoga and Abeta). A total of 38 samples were collected from the local markets in 7 subcounties in Homa Bay County, Kenya. The levels of aflatoxin were determined using an indirect competitive ELISA protocol. Yeast and mould contamination was determined using ISO 21527-2 method. Mean aflatoxin levels in chopped, fermented, and sun-dried cassava (Akuoga) were 0.36 μg/kg compared to 0.25 μg/kg in chopped and sun-dried (Abeta) products. Aflatoxin contamination was detected in 55% of the samples and ranged from 0–5.33 μg/kg. These levels are within 10 μg/kg recommended by the CODEX STAN 193-1995. Yeast and mould counts in fermented and chopped sun-dried products were 3.16 log Cfu/g and 2.92 log Cfu/g, respectively. The yeast and mould counts were above standards set by East African Standard 739:2010 in 62% (Akuoga) and 58% (Abeta). The most prevalent fungal species were Saccharomyces cerevisiae (68.4%) and Candida rugosa (68%) followed by Candida parapsilosis (18.4%), Candida tropicalis (15.8%), Candida humilis (15.8%), and Aspergillus spp. (5.3%). Aspergillus spp. was the only mycotoxigenic mould isolated from the samples. The study shows that cassava consumers are exposed to the risk of aflatoxin poisoning. The study, therefore, recommends appropriate surveillance to ensure safety standards.

Description: Efficient conversion of pentose sugars to ethanol is important for an economically viable lignocellulosic bioethanol process. Ten yeasts fermenting both D-xylose and L-arabinose were subjected to an adaptation process with L-arabinose as carbon source in a medium containing acetic acid. Four Meyerozyma caribbica-adapted strains were able to ferment L-arabinose to ethanol in the presence of 3 g/L acetic acid at 35°C. Meyerozyma caribbica Mu 2.2f fermented L-arabinose to produce 3.0 g/L ethanol compared to the parental strain with 1.0 g/L ethanol in the absence of acetic acid. The adapted M. caribbica Mu 2.2f strain produced 3.6 and 0.8 g/L ethanol on L-arabinose and D-xylose, respectively, in the presence of acetic acid while the parental strain failed to grow. In a bioreactor, the adapted M. caribbica Mu 2.2f strain produced 5.7 g/L ethanol in the presence of 3 g/L acetic acid with an ethanol yield and productivity of 0.338 g/g and 0.158 g/L/h, respectively, at a KLa value of 3.3 h−1. The adapted strain produced 26.7 g/L L-arabitol with a yield of 0.900 g/g at a KLa value of 4.9 h−1.

Description: Streptococcus pyogenes are associated with many bacterial diseases in both humans and animals and are capable of causing a multitude of human diseases. S. pyogenes isolates were identified by their bacitracin sensitivity, positive spy1258 detection, and positive GAS latex agglutination. Different isolates were typed serotypically and genotypically by BOX-PCR. Different virulence factors were identified in S. pyogenes isolates. In addition, antimicrobial resistance was tested to eleven different antibiotics. Furthermore, the resistance mechanisms were determined phenotypically by the disc diffusion method. Finally, the correlation between both molecular and serotypes identified and the profile of virulence factors and clinical and geographical sources was determined for all isolates. Thirty-eight S. pyogenes isolates were collected from different clinical sources. Resistance testing indicated high resistance to mostly used antibiotics except amoxicillin/clavulanic acid, amoxicillin, and ampicillin. Serotyping results indicated five different serotypes, M1, M2, M3, M4, and M6, in S. pyogenes isolates, while six isolates were identified as untypeable. In addition, positive PCR results identified most of the tested SAgs genes in which speJ gene was mostly identified followed by speI, speC, and ssa genes being identified in 81.6%, 63.3%, 60.5%, and 60.5%, respectively. However, speH was the least detected. In contrast, speL, speM, and smeZ genes could not be detected in all tested isolates. Finally, BOX-PCR molecular typing was a more effective clustering method when compared to the serotyping method in all S. pyogenes. In conclusion, the isolates in this study were highly resistant to mostly used antibiotics. M1 was the most identified serotype. No significant association was found between serotypes, BOX-PCR cluster groups, and SAgs genes profiles. However, by the application of BOX-PCR, effective molecular typing was obtained.

Description: Background. Asymptomatic bacteriuria is one of the major risk factors for the development of urinary tract infections during pregnancy which accounts for about 70% of the cases. However, there is no guideline which recommends routine screening of pregnant women for asymptomatic bacteriuria in most of developing countries including Ethiopia. Thus, the aim of this study was to determine the magnitude, associated factors, and antimicrobial susceptibility pattern of asymptomatic bacteriuria among pregnant women. Methods. A cross-sectional study was conducted from March to April 2019. Data were collected through face-to-face interview and analyzed using Statistical Package for Social Science version 22. A test of association was performed using logistic regression and P value less than 0.05 was considered statistically significant. Results. The overall prevalence of asymptomatic bacteriuria was 19.9%. Direction of wiping after genital wash, postcoital urination, and catheterization were factors significantly associated with asymptomatic bacteriuria. Most of the isolated Gram positive were highly sensitive to Ceftriaxone (90.9%). Coagulase-negative staphylococci showed higher sensitivity to Augmentin (75.0) and Ceftriaxone (87.5%), whereas they showed resistance to Clindamycin (68.7%) and Ampicillin (62.5%). Gram-negative bacteria isolates showed higher sensitivity to Ceftriaxone (88.2%), Gentamycin (67.5%), and Augmentin (64.7%), while they showed resistance to Ampicillin (70.5%) and Clindamycin (50.0%). Conclusion. The overall prevalence of asymptomatic bacteriuria among pregnant women in this study was high. Direction of wiping after genital wash, catheterization, and postcoital urination increases the odds of asymptomatic bacteriuria. Therefore, health education on the predisposing factors is strongly recommended.

Description: Thirty-three (33) isolates of methicillin-resistant Staphylococcus aureus (MRSA) from healthy edible marine fish harvested from two aquaculture settings and the Kariega estuary, South Africa, were characterised in this study. The phenotypic antimicrobial susceptibility profiles to 13 antibiotics were determined, and their antibiotic resistance determinants were assessed. A multiplex PCR was used to determine the epidemiological groups based on the type of SCCmec carriage followed by the detection of staphylococcal enterotoxin-encoding genes sea-sed and the Panton Valentine leucocidin gene (pvl). A high antibiotic resistance percentage (67–81%) was observed for Erythromycin, Ampicillin, Rifampicin, and Clindamycin, while maximum susceptibility to Chloramphenicol (100%), Imipenem (100%), and Ciprofloxacin (94%) was recorded. Nineteen (58%) of the MRSA strains had Vancomycin MICs of ≤2 μg/mL, 4 (12%) with MICs ranging from 4–8 μg/mL, and 10 (30%) with values ≥16 μg/mL. Overall, 27 (82%) isolates were multidrug-resistant (MDR) with Erythromycin-Ampicillin-Rifampicin-Clindamycin (E-AMP-RIP-CD) found to be the dominant antibiotic-resistance phenotype observed in 4 isolates. Resistance genes such as tetM, tetA, ermB, blaZ, and femA were detected in two or more resistant strains. A total of 19 (58%) MRSA strains possessed SCCmec types I, II, or III elements, characteristic of healthcare-associated MRSA (HA-MRSA), while 10 (30%) isolates displayed SCCmec type IVc, characteristic of community-associated MRSA (CA-MRSA). Six (18%) of the multidrug-resistant strains of MRSA were enterotoxigenic, harbouring the see, sea, or sec genes. A prevalence of 18% (6/33) was also recorded for the luk-PVL gene. The findings of this study showed that marine fish contained MDR-MRSA strains that harbour SCCmec types, characteristic of either HA-MRSA or CA-MRSA, but with a low prevalence of enterotoxin and pvl genes. Thus, there is a need for continuous monitoring and implementation of better control strategies within the food chain to minimise contamination of fish with MDR-MRSA and the ultimate spread of the bug.

Description: This work aimed to retrieve a field isolate of probiotic from Nile tilapia (Oreochromis niloticus) and compare the obtained results with a commercial probiotic product through experimental studies. The study was conducted on 250 Nile tilapia. Ten fish were used to isolate the probiotic strain. Two isolates showed an in vitro inhibitory effect against pathogenic A. hydrophila. The isolate with the largest zone was identified by PCR. Sixty fish were used to test the safety of a potential probiotic. One hundred and eighty fish were used in a two-month feeding experiment. Fish were divided into 3 groups, group (1): the control, group (2): fed on potential probiotics, and group (3): fed on commercial probiotic (Organic Green™). The effects of tested products on the immune response were recorded in all groups. After one and two months of feeding experiment, blood and nonspecific immune parameters were evaluated. Disease resistance against Aeromonas hydrophila was evaluated through challenge experiment. The histopathology of the treated groups was fully recorded in comparison with the control group. The potential probiotic based on the in vitro antimicrobial activity test was identified as P. putida using routine and gel electrophoresis and 16S rRNA sequencing. During the first and the second month of experiment, there was a highly significant increase in the survival percent of the experimental fish in both treated groups with probiotics. In the first phase of the experiment, a significant increase in the haematocrit values and NBT, lysozyme activity, and phagocytic activity was seen in all treated groups in comparison with the control. The increase in the TLC was significant in the group fed with P. putida in comparison with the control group. In the second phase, a nonsignificant increase in the hematocrit values and significant increases in the NBT and phagocytic index were seen in P. putida and organic green groups in comparison with the control group. The TLC and DLC revealed nonsignificant changes in the treated groups in comparison with the control. The RLP in the groups treated with P. putida was higher than that in those treated with organic green. Although probiotics are an important management tool in aquaculture, it should be subjected to scientific laboratory tests and field measurements.

Description: Optimal recovery of mycobacteria from the contaminated liquid culture is a challenge. While alternative methods have been suggested to reduce the rate of contamination in the BACTEC MGIT 960 system, reprocessing the contaminated liquid culture improves recovery of Mycobacterium tuberculosis. Among 793 MGIT cultures raised from as many sputum specimens after primary decontamination by the standard NaLC-NaOH method, valid results were available for 687 (86.6%) as 106 (13.4%) were contaminated. Reprocessing and reculturing of the contaminated cultures increased valid results to 739 (93.2%) and reduced the contamination rate to 6.8%. Both values were statistically significant. Recovery of the Mycobacterium tuberculosis complex increased from 45.6% to 48.4%. Valid negative results were available for an additional 3.4%. The method may be adopted to reduce the rate of contamination and to improve the valid culture results for mycobacteria.

Description: Although the epidemiology of pathogenic Candida species is changing due to invasive diseases, Candida albicans has become the common cause of human infections worldwide. Candida albicans is a diploid yeast with a mostly clonal mode of reproduction and without known complete sexual cycle. This species has two heterozygous and homozygous strains at hyphal wall protein 1 gene locus (hwp1). Little is known about virulence factors of these strains. The aim of this study was to evaluate the exoenzyme activity of heterozygous and homozygous C. albicans strains. A total of 60 stock Candida albicans species isolates, which consisted of 30 homozygous and 30 heterozygous strains, were used for exoenzyme activities. We used egg yolk agar, Sabouraud blood agar, and bovine serum albumin agar for evaluation of phospholipase, hemolysin, and proteinase activity, respectively. Homozygous strains of Candida albicans had more phospholipase and proteinase activity than heterozygous strains. However, there were no significant statistical differences between the two strains in the severity of exoenzymes production. Beta hemolysin activity was seen in 100% and 96.7% of the homozygous and heterozygous strains, respectively. The results of this study indicated that both of the strains exhibited exoenzyme activities in different ranges. There were no significant statistical differences in virulence factors between the homozygous and heterozygous strains.

Description: The human microbiome comprises bacteria, archaea, viruses, and eukaryotes which reside within and outside our bodies. These organisms impact human physiology, both in health and in disease, contributing to the enhancement or impairment of metabolic and immune functions. Micro-organisms colonise various sites on and in the human body, where they adapt to specific features of each niche. Facultative anaerobes are more dominant in the gastrointestinal tract, whereas strict aerobes inhabit the respiratory tract, nasal cavity, and skin surface. The indigenous organisms in the human body are well adapted to the immune system, due to the biological interaction of the organisms with the immune system over time. An alteration in the intestinal microbial community plays a major role in human health and disease pathogenesis. These alterations result from lifestyle and the presence of an underlying disease. Dysbiosis increases host susceptibility to infection, and the nature of which depends on the anatomical site involved. The unique diversity of the human microbiota accounts for the specific metabolic activities and functions of these micro-organisms within each body site. It is therefore important to understand the microbial composition and activities of the human microbiome as they contribute to health and disease.

Description: Many diseases have been associated with poor drinking water quality including diseases caused by diarrheagenic pathogens, especially in developing countries where access to a consistent water supply is a problem. The objective of the study was to evaluate the health risks associated with the sources of drinking water in the Dangme West District of Ghana using E. coli as a measurement tool, aiming at ascertaining the paths leading to contamination of the water sources. A total of 464 water samples were obtained for analysis. Sampling covered a year across the dry and wet seasons in Ghana. Water samples were obtained from groundwater and surface water sources. E. coli counts were determined using the most probable number method (MPN). Disease risk assessment was carried out using the WHO drinking water risk assessment guidelines. Generally, the study revealed significantly higher E. coli counts in the wet season than in the dry season. Among the water samples analyzed, surface water, especially from the dams, was found to pose the highest disease risk as compared to the other water sources. Samples from groundwater sources, especially boreholes, posed the lowest disease risk. In conclusion, observations from the study implied that most water sources in the study district are highly polluted with bacteria pathogens beyond recommended safety guidelines. The main causes of faecal contamination in these water sources were purported to be anthropogenic. Therefore, there is a need to formulate a policy aimed at managing and improving rural water sources.

Description: The biofilm formation on the surfaces which are in direct contact with food products might lead to their contamination and, consequently, present serious health problems for the consumers. The goals of the present work were to study P. aeruginosa biofilm formation on two granites and to investigate the efficiency of sodium hypochlorite (NaCLO) against the same biofilm formed on these substrata using the plate count method (PCM) and epifluorescence microscopy (EP). More biofilm cells adhered to Rosa Porrino than Gris Pinhel, and the PCM method indicated that NaCLO was efficient against the biofilm installed on the Gris Pinhel at the concentration of 1.5% after 15 min of treatment, while it was not efficient against the one installed on the Rosa Porrino. By contrast, the EP showed that the biofilm persists on two granites after NaCLO treatment, at different concentrations and contact times. In addition, the surface properties of granites such as mineral composition, roughness, and physicochemical properties were determined by X-ray diffraction (XRD), scanning electron microscopy coupled with electron diffraction spectroscopy coupled with energy-dispersive X-ray spectroscopy (SEM-EDS), Fourier transform infrared (FTIR), atomic force microscopy (AFM), and contact angle measurement (CAM), respectively. The results revealed that Gris Pinhel is hydrophilic with a high roughness value and Rosa Porrino is hydrophobic with low roughness, while both of them contain the quartz, feldspar, and mica as the main dominant compositions.

Description: Molecular detection and classification of the bacterial groups in a sample are relevant in several areas, including medical research and forensics. Sanger sequencing of the 16S rRNA gene is considered the gold standard for microbial phylogenetic analysis. However, the development of massively parallel sequencing (MPS) offers enhanced sensitivity and specificity for microbiological analyses. In addition, 16S rRNA target amplification followed by MPS facilitates the combined use of multiple markers/regions, better discrimination of sample background, and higher sample throughput. We designed a novel set of 16S rRNA gene primers for detection of bacterial species associated with clinical, bioweapon, and biohazards microorganisms via alignment of 364 sequences representing 19 bacterial species and strains relevant to medical and forensics applications. In silico results indicated that the hypervariable regions (V1V2), (V4V5), and (V6V7V8) support the resolution of a selected group of bacteria. Interspecies and intraspecies comparisons showed 74.23%–85.51% and 94.48%–99.98% sequencing variation among species and strains, respectively. Sequence reads from a simulated scenario of bacterial species mapped to each of the three hypervariable regions of the respective species with different affinities. The minimum limit of detection was achieved using two different MPS platforms. This protocol can be used to detect or monitor as low as 2,000 genome equivalents of bacterial species associated with clinical, bioweapon, and biohazard microorganisms and potentially can distinguish natural outbreaks of pathogenic microorganisms from those occurring by intentional release.

Description: An outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first reported in Wuhan City, China, in December 2019. Since then, the outbreak has grown into a global pandemic, and neither a vaccine nor a treatment for the disease, termed coronavirus disease 2019 (COVID-19), is currently available. The slow translational progress in the field of research suggests that a large number of studies are urgently required. In this context, this review explores the impact of bacteriophages on SARS-CoV-2, especially concerning phage therapy (PT). Bacteriophages are viruses that infect and kill bacterial cells. Several studies have confirmed that in addition to their antibacterial abilities, bacteriophages also show antiviral and antifungal properties. It has also been shown that PT is effective for building immunity against viral pathogens by reducing the activation of NF kappa B; additionally, phages produce the antiviral protein phagicin. The Ganges river in India, which originates from the Himalayan range, is known to harbor a large number of bacteriophages, which are released into the river gradually by the melting permafrost. Water from this river has traditionally been considered a therapeutic agent for several diseases. In this review, we hypothesize that the Ganges river may play a therapeutic role in the treatment of COVID-19.

Description: Streptomyces has been reported as an essential producer of bioactive substances, including antibiotics and other types of antimicrobials. This study investigated antibacterial-producing Streptomyces isolated from the gut of estuarine fish Chanos chanos, emphasizing screening for the producer of peptide-containing antibacterial compounds. Eighteen isolates were found during preliminary screening, in which four isolates showed the best antibacterial activities. Based on the morphological, physiological, and biochemical characterization, as well as 16S rRNA partial sequencing, all of the four isolates belonged to Streptomyces. Three isolates were suspected as novel isolate candidates based on homology presentations and phylogenetic tree analysis. Disk-diffusion assay of the metabolite-crude-extract from the isolates showed broad-spectrum inhibitory activities against Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 10876, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa InaCC B52 with minimum inhibitory concentration and minimum bactericidal concentration ranging from 2.5–10 mg/mL and 5–10 mg/mL, respectively. The highest antibacterial activity with low MIC and MBC values was shown by isolate AIA-10. Qualitative HPLC profiling revealed that the metabolic-crude-extracts showed many peaks with intensive area at 210 and 214 nm, especially from SCA-11 and AIA-10, indicating the presence of peptide groups in the structure of the constituent compound. The results also suggested that crude extracts SCA-11 and AIA-10 had higher hydrophobicity properties than the other extracts. Further characterization of the active compound was needed to find out which compounds were responsible for the antibacterial activity. The results of this study indicated that some Streptomyces isolated from new environmental niches, i.e., gut of estuarine fish Chanos chanos, produce promising peptide-containing bioactive compounds.

Description: High extrusion temperatures may compromise the functionality of probiotics in dry food. This study aimed to (i) evaluate the effects of two types of microencapsulation techniques, different encapsulating agents, and 120 days of storage on the viability of a commercial probiotic product and (ii) investigate fecal microbiota populations and fecal characteristics of adult cats fed with diets supplemented with probiotics. Three experimental treatments were evaluated: T1, commercial feed (control); T2, commercial kibbles coated with probiotics; and T3, commercial feed supplemented with freeze-dried probiotics and fructooligosaccharides. Fructooligosaccharides and gum arabic were used as encapsulating agents for freeze drying and spray drying and a pool containing Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus lactis, Bifidobacterium bifidum, Enterococcus faecium, and Saccharomyces cerevisiae as a probiotic. Diets were provided to 18 adult cats for 20 days. Feed samples were evaluated microbiologically, and feces were characterized according to their microbial content, pH, and fecal score. Freeze drying was more effective in maintaining microbial viability. Microcapsules prepared using fructooligosaccharides as encapsulants had the highest bacterial count: 8.74 log CFU/g of lactic acid bacteria and 8.75 log CFU/g of enterococci. Probiotics and synbiotics positively modulated (P

Description: Urinary tract infection (UTI) is one of the most prevalent bacterial infections in the world affecting the bladder and the kidney. Escherichia coli (E. coli) is the main causative agent of 80–90% of community-acquired UTIs, about 40% of nosocomial UTIs, and 25% of recurrent UTIs. The field of proteomics has emerged as a great tool to analyze expressed proteins to identify possible biomarkers associated with many pathological states and, to the same extent, those associated with bacterial pathogenesis and their ability to cause recurrent infections. Here, in a descriptive cross-sectional pilot study, we employed proteomic techniques to investigate the effects of environmental stress on protein profiles of E. coli simulated by sequential passaging of samples from patients with UTIs to screen for unique proteins that arise under stressful environment and could aid in the early detection of UTIs. Four urine samples were collected from individuals with recurrent UTI and sequentially subcultured; protein samples were extracted from bacterial pellets and analyzed using 2-dimensional gel electrophoresis (2DGE). Protein spots of interest arising from changes in the protein profile were analyzed using liquid chromatography-mass spectrometry (LC-MS/MS) and matched against known databases to identify related proteins. We identified ATPB_ECOBW, ASPA ECOLI, DPS ECOL6, and DCEB ECOLI as proteins associated with higher passaging. We concluded that passaging resulted in identifiable changes in the protein profile of E. coli, namely, proteins that are associated with survival and possible adaptation of bacteria, suggestive of factors contributing to antibiotic resistance and recurrent UTIs. Furthermore, our method could be further used to identify indicator-protein candidates that could be a part of a growing protein database to diagnose and identify causative agents in UTIs.

Description: Understanding the honeybee gut bacteria is an essential aspect as honeybees are the primary pollinators of many crops. In this study, the honeybee-associated gut bacteria were investigated by targeting the V3-V4 region of 16S rRNA genes using the Illumina MiSeq. The adult worker was captured in an urban area in a dense settlement. In total, 83,018 reads were obtained, revealing six phyla from 749 bacterial operational taxonomic units (OTUs). The gut was dominated by Proteobacteria (58% of the total reads, including Enterobacteriaceae 28.2%, Erwinia 6.43%, and Klebsiella 4.90%), Firmicutes (29% of the total reads, including Lactococcus garvieae 13.45%, Lactobacillus spp. 8.19%, and Enterococcus spp. 4.47%), and Actinobacteria (8% of the total reads, including Bifidobacterium spp. 7.96%). Many of these bacteria belong to the group of lactic acid bacteria (LAB), which was claimed to be composed of beneficial bacteria involved in maintaining a healthy host. The honeybee was identified as Apis nigrocincta based on an identity BLAST search of its COI region. This study is the first report on the gut microbial community structure and composition of A. nigrocincta from Indonesia.

Description: Aspergillus nidulans is a filamentous fungus that is a potential resource for industrial enzymes. It is a versatile fungal cell factory that can synthesize various industrial enzymes such as cellulases, β-glucosidases, hemicellulases, laccases, lipases, proteases, β-galactosidases, tannases, keratinase, cutinases, and aryl alcohol oxidase. A. nidulans has shown the potential to utilize low-cost substrates such as wheat bran, rice straw, sugarcane bagasse, rice bran, coir pith, black gram residue, and chicken feathers to produce enzymes cost-effectively. A. nidulans has also been known as a model organism for the production of heterologous enzymes. Several studies reported genetically engineered strains of A. nidulans for the production of different enzymes. Native as well as heterologous enzymes of A. nidulans have been employed for various industrial processes.

Description: Steered fermentation by microorganisms gives great added value in the nutritional quality of local food. Ginger rhizome naturally contains a myriad of bioactive compounds including polyphenol and flavonoids. The aim of this work was to ferment the ginger juice, to evaluate the biochemical parameters of ginger wine, and to understand the involvement of microorganisms in the bioincrease of polyphenol compounds. Titratable acidity and pH values were determined and showed that pH is around 1.6 at the end of the fermentation when the acidity is around 6.431 g/L. Using colorimetric assay, the total polyphenolic and flavonoid compounds were evaluated throughout the fermentation. The variation of the polyphenol and flavonoid concentrations of the unsweetened sample was around 10.18 to 14.64 mg Eq AG/g and 1.394 to 2.224 mg Eq Cat/g Ms, but those from the sweet sample were around 10.82 to 18.34 mg Eq AG/g Ms and 1.311 to 2.290 mg Eq Cat/g. Using one-step PCR, multiplex techniques with specific primers, with yeast-like phenotype 27.27% (6), have been assigned among 22 isolates to Saccharomyces cerevisiae. By using PCR multiplex techniques, Bacillus licheniformis, Bacillus pumilus, Bacillus safensis, and Saccharomyces cerevisiae have been identified. Together with Saccharomyces cerevisiae, we showed that Bacillus sp. are able to secrete enzymatic landscape with some activities up to 50% including cellulase, amylase, pectinase, and protease.

Description: Background. Urinary tract infection is a major public health problem in terms of morbidity and mortality worldwide. It ranks as the number one infection which leads to an antibiotic prescription after a physician’s visit. However, there are limited studies done on UTI in Ethiopia. Hence, this study was aimed to assess the magnitude of urinary tract infection and its associated factors among adult patients attending hospitals of the Tigray region, Ethiopia. Methods and Material. A hospital-based cross-sectional study was conducted from April to May 2019. Systematic random sampling technique was used to select 472 participants from five randomly selected hospitals in Tigray region. A pretested structured questionnaire through face-to-face interview and patient chart review checklist was used to collect data. Data were analyzed by SPSS version 21. A binary logistic regression model was used to test the association between dependent and independent variables. Result. The magnitude of urinary tract infection was 86 (18.2%) (95% CI: 14.6%–21.6%). After adjustment of the independent variables, the significant factors associated with urinary tract infection were being female (AOR = 3.50; 95% CI: 1.88–6.51), urine passing frequency 

Description: This study was conducted to detect the presence of colistin-resistant Escherichia coli (E. coli) in raw chicken meat and bean sprouts collected from local markets and to determine the antimicrobial resistance patterns of the E. coli isolates. A total of 100 samples, comprised of 50 raw chicken meat and 50 bean sprouts, were collected and processed. Kirby-Bauer method was used to determine the antimicrobial resistance patterns, and PCR amplification was used to detect E. coli species-specific and colistin resistance (mcr-1 and mcr-2) genes. The results showed that 52.1% (12/23) of the E. coli isolated from raw chicken meat were positive for the colistin resistance encoding gene, mcr-1, whereas all the E. coli isolates from bean sprouts were negative for colistin resistance encoding genes. The findings show that chicken meat contaminated with colistin-resistant E. coli may pose public health risk to the consumers. Hence, prudent usage of antibiotics and hygienic handling of food items helps to prevent and combat the risks of spreading of colistin-resistant E. coli and the public health risks it may pose. More comprehensive and large-scale studies focusing on all the possible sources of colistin-resistant E. coli are recommended.

Description: The aim of this study was to characterise Vibrio species of water samples collected from taps, boreholes, and dams in the North West province, South Africa, and assess biocontrol potentials of their bacteriophages. Fifty-seven putative Vibrio isolates were obtained on thiosulfate-citrate-bile-salt-sucrose agar and identified using biochemical tests and species-specific PCRs. Isolates were further characterised based on the presence of virulence factors, susceptibility to eleven antibiotics, and biofilm formation potentials. Twenty-two (38.60%) isolates were confirmed as Vibrio species, comprising V. harveyi (45.5%, n = 10), V. parahaemolyticus (22.7%, n = 5), V. cholerae (13.6%, n = 3), V. mimicus (9.1%, n = 2), and V. vulnificus (9.1%, n = 2). Three of the six virulent genes screened were positively amplified; four V. parahaemolyticus possessed the tdh (18.18%) and trh (18.18%) genes, while the zot gene was harboured by 3 V. cholerae (13.64%) and one V. mimicus (4.55%) isolate. Isolates revealed high levels of resistance to cephalothin (95.45%), ampicillin (77.27%), and streptomycin (40.91%), while lower resistances (4.55%–27.27%) were recorded for other antimicrobials. Sixteen (72.7%) isolates displayed multiple antibiotic-resistant properties. Cluster analysis of antibiotic resistance revealed a closer relationship between Vibrio isolates from different sampling sites. The Vibrio species displayed biofilm formation potentials at 37°C (63.6, n = 14), 35°C (50%, n = 11), and 25°C (36.4%, n = 8). Two phages isolated in this study (vB_VpM_SA3V and vB_VcM_SA3V) were classified as belonging to the family Myoviridae based on electron microscopy. These were able to lyse multidrug-resistant V. parahaemolyticus and V. cholerae strains. These findings not only indicate the presence of antibiotic-resistant virulent Vibrio species from dam, borehole, and tap water samples that could pose a health risk to humans who either come in contact with or consume water but also present these lytic phages as alternative agents that can be exploited for biological control of these pathogenic strains.

Description: The purpose of this study was to establish the probiotic potential of lactic acid bacteria (LAB) starter cultures, Lb. plantarum MNC 21, L. lactis MNC 24, and W. confusa MNC 20, isolated from a traditionally fermented sorghum-millet beverage from Uganda. The cultures were examined for tolerance to acid and bile salts, bile salt hydrolase (BSH) activity, antibiotic susceptibility, biogenic amine production, mucin degradation, hydrophobicity, auto-aggregation, adherence to the ileum, coaggregation, and antimicrobial properties against selected pathogenic species. Lb. rhamnosus yoba 2012, a known probiotic, was the reference. The isolates were tolerant to acid (pH = 3) and bile (1%). W. confusa MNC 20 and Lb. plantarum MNC 21 exhibited medium BSH activity (11–15 mm diameter of hydrolysis zone) while L. lactis and Lb. rhamnosus yoba 2012 exhibited low BSH activity (

Description: Background. Vaginal or genitourinary infections are a major cause of morbidity, sterility, and increase in the vulnerability to cancers and HIV/AIDS infection. The aim of this study was to determine the prevalence of vaginal infections of C. albicans, G. vaginalis, and T. vaginalis among women in the locality of Dschang, West Region of Cameroon. Method. A prospective study was carried out in the District Hospital of Dschang. After obtaining informed consent, one thousand and one (1001) samples of vaginal swabs were collected. Biological diagnosis was carried out on fresh samples, Gram stained, and then cultivated in Sabouraud agar in a Petri dish, in order to isolate and identify the various infectious agents. Results. Five hundred and twenty-five (525) women were diagnosed positive, hosting at least one of these microorganisms, making an overall prevalence of 52.44%. Two hundred and fifty-six (256) women (25.57%) were infected with C. albicans, and 171 (17.08%) with G. vaginalis. Ninety-five (9.49%) were infected with both C. albicans and G. vaginalis, 2 (0.20%) with C. albicans and T. vaginalis, and 1 (0.1%) with G. vaginalis and T. vaginalis. Conclusion. Drastic measures should be taken in order to improve life styles to regress the frequency of these infections. Results obtained in this study, will help to educate and shed more light on the prevalence of vaginal infections in the West Region of Cameroon.

Description: Peanut paste produced in multipurpose mills is very often the site of choice for fungal contaminants that pose a major risk to consumers. The objective of this study is to evaluate the level of fungal contamination of peanut paste produced according to different moulding processes during storage. Thirty samples of peanut paste were produced from 60 kg of peanut pods according to three types of moulding (domestic moulding, artisanal moulding, and hygienic moulding) and then preserved for three months. These thirty samples were subjected to microbiological analysis using the conventional mould count method. The moisture content of the various peanut pastes was determined according to the AOAC method. Fungi were identified by using taxonomic schemes based on microscopic observation and culture appearance. Mould loads ranged from 0 to 6.4.102 cfu/g; 91 to 9.6.102 cfu/g; and 0 to 4.6.102 cfu/g, respectively, for domestic, artisanal, and hygienic mouldings during conservation. Moisture content increases during the conservation of peanut paste. It increases from 1.23 to 3.17% for domestic moulding, 1.30 to 3.20% for artisanal moulding, and 1.30 to 2.94% for hygienic moulding. Four fungal genera, namely, Aspergillus, Mucor, Absidia, and Penicillium and three species of Aspergillus including A. flavus, A. fumigatus and A. niger have been identified. The peanut paste produced from domestic and hygienic moulding is less contaminated during storage than that obtained in the artisanal way.

Description: Although what unifies the carcinogenic microorganisms has not been determined by multiple studies, the role of bacteria in the development of neoplasms has not been properly elucidated. In this review, we discuss links between the bacterial species and cancer, with focus on immune responses for the stimulation of tumor cells such as induction of inflammation. Finally, we will describe the potential therapeutic strategies of bacteria on target tumors to improve treatment while mitigating adverse reactions. Cancer is a series of genetic changes that transform normal cells into tumor cells. These changes come from several reasons, including smoking, drinking alcohol, sunlight, exposure to chemical or physical factors, and finally chronic infection with microorganisms, including bacteria. In fact, bacterial infections are not carcinogenic, but recently it was discovered that the association between bacteria and cancer is through two mechanisms, the first stimulating chronic inflammation and the second producing carcinogenic metabolites. While bacteria are carcinogenic agents also, they have a dual role eliminating and removing tumor cells. However, the traditional cancer treatments that include chemotherapy, radiotherapy, surgery, and immunotherapy increase the chances of survival, and there are many side effects of these therapies, including the high toxicity of tissues and normal cells, could not penetrate the tumor cells, and resistance of these therapies by tumor cells. Therefore, the world has turned to an alternative solution, which is the use of genetically engineered microorganisms; thus, the use of living bacteria targeting cancerous cells is the unique option to overcome these challenges. Bacterial therapies, whether used alone or combination with chemotherapy, give a positive effect to treat multiple conditions of cancer. Also, bacteria can be used as vectors for drug, gene, or therapy, and this is a great step to treat cancer. Thus, we review the mechanisms underlying the interaction of the microbiota residents with cancer. Cancer-associated bacteria differ from those in healthy human and are linked with gene-expression profile. We also discuss how live bacteria interact with tumor microenvironments to induce tumor regression through colonization and spread. Finally, we provide past and ongoing clinical trials that include bacteria targeting tumors.

Description: Newcastle disease (ND) causes significant economic losses in the poultry industry in developing countries. In Kenya, despite rampant annual ND outbreaks, implementation of control strategies is hampered by a lack of adequate knowledge on the circulating and outbreak causing-NDV strains. This study reports the first complete genome sequences of NDV from backyard chicken in Kenya. The results showed that all three isolates are virulent, as assessed by the mean death time (MDT) and intracerebral pathogenicity index (ICPI) in specific antibody negative (SAN) embryonated eggs and 10-day-old chickens, respectively. Also, the polybasic amino acid sequence at the fusion-protein cleavage site had the motif 112RRQKRFV118. Histopathological findings in four-week-old SPF chicken challenged with the NDV isolates KE001, KE0811, and KE0698 showed multiple organ involvement at five days after infection with severe effects seen in lymphoid tissues and blood vessels. Analysis of genome sequences obtained from the three isolates showed that they were 15192 base pair (bp) in length and had genomic features consistent with other NDV strains, the functional sites within the coding sequence being highly conserved in the sequence of the three isolates. Amino acid residues and substitutions in the structural proteins of the three isolates were similar to the newly isolated Tanzanian NDV strain (Mbeya/MT15). A similarity matrix showed a high similarity of the isolates to NDV strains of class II genotype V (89–90%) and subgenotype Vd (95–97%). Phylogenetic analysis confirmed that the three isolates are closely related to NDV genotype V strains but form a distinct cluster together with NDV strains from the East African countries of Uganda and Tanzania to form the newly characterized subgenotype Vd. Our study provides the first description of the genomic and pathological characteristics of NDV of subgenotype Vd and lays a baseline in understanding the evolutionary dynamics of NDV and, in particular, Genotype V. This information will be useful in the development of specific markers for detection of viruses of genotype V and generation of genotype matched vaccines.

Description: To investigate the effect of flour and starch of the Indonesian native tuber “taro” on the composition and activity of the gut microbiota in diabetic rats, streptozotocin (STZ)-induced diabetic rats were fed normal chow (AIN), or AIN in which corn starch was replaced by either taro flour or purified taro starch for 4 weeks. Fecal samples were collected at baseline and after 4 weeks, and the composition of microbial communities was measured using 16S rRNA sequencing, while SCFAs were measured using ion chromatography. Bodyweight declined upon DM induction with STZ. Feeding taro starch led to a lower reduction in bodyweight than feeding taro starch, but this was only significant for taro starch in weeks 2, 3, and 4 (p=0.02, p=0.01, and p

Description: Escherichia coli is related to foodborne disease and outbreaks worldwide. It mainly affects persons at high risk as newborns, infants, and individuals with impaired immune system in hospitals. Multidrug-resistant E. coli is currently spreading both in community and hospital settings. Our study aims to evaluate the presence of E. coli and the incidence of its antibiotic resistance in samples obtained from various cooked and raw foods (N = 300), food contact surfaces (N = 238), and food handlers (N = 40) in Moroccan hospital catering service. E. coli was identified using API 20E, and the antibiotic resistance patterns were obtained using the agar disk diffusion methods. However, PCR method was used for O157 and H7 typing. The samples analysis showed that 14.33%, 24.16%, and 45% of food, surfaces, and food handlers harbored E. coli, respectively, with the highest rates obtained in raw meats (34.88%) and salads (34.88%). Molecular amplification shows that 14 E. coli isolates carried the flagellar antigen H7, while there are no isolates showing amplification for O157. The high rate of resistance was noted against ampicillin (100%), amoxicillin-clavulanate acid (100%), nalidixic acid (61.62%), and cefotaxime (59.49%), and isolates obtained from food handler’s hands showed the highest rates of resistance. None of the isolates are extended-spectrum beta-lactamases producing, while 27.7% of the isolates were metallo-beta-lactams producing. This first study conducted on Moroccan hospital catering services may draw the authorities’ attention to the necessity of setting up a surveillance system to monitor the food preparation process and the safety of prepared food in healthcare settings.

Description: Crude oil pollution has consistently deteriorated all environmental compartments through the cycle of activities of the oil and gas industries. However, there is a growing need to identify microbes with catabolic potentials to degrade these pollutants. This research was conducted to identify bacteria with functional degradative genes. A crude oil-polluted soil sample was obtained from an aged spill site at Imo River, Ebubu, Komkom community, Nigeria. Bacteria isolates were obtained and screened for hydrocarbon degradation potential by turbidometry assay. Plasmid and chromosomal DNA of the potential degraders were further screened for the presence of selected catabolic genes (C230, Alma, Alkb, nahAC, and PAHRHD(GP)) and identified by molecular typing. Sixteen (16) out of the fifty (50) isolates obtained showed biodegradation activity in a liquid broth medium at varying levels. Bacillus cereus showed highest potential for this assay with an optical density of 2.450 @ 600 nm wavelength. Diverse catabolic genes resident in plasmids and chromosomes of the isolates and, in some cases, both plasmid and chromosomes of the same organism were observed. The C230 gene was resident in 〉50% of the microbial population tested, while other genes occurred in lower proportions with the least observed in nahAC and PAHRHD. These organisms can serve as potential bioremediation agents.

Description: This study aims to detect Staphylococcus aureus (S. aureus) resistance in the veterinary hospital environment. S. aureus are one of the components of the microbiota, and they may be present in patients in a veterinary hospital environment. Methicillin resistance is determined by a chromosomal gene (mecA), which codes for modifications in the beta-lactam antibiotic receptor, where the penicillin-binding protein will have a low affinity for the antibiotic. Samples were collected through swabs of materials and equipment at the hospital. S. aureus was identified in 7.6% (21/276) of the samples collected, and of the 21 strains isolated, 4 (19.0%) carried the mecA gene. MRSA are all strains of S. aureus that express the mecA gene. Four strains harbor the mecA gene; however, only two expressed the phenotypic resistance to cefoxitin and were characterized as MRSA. An isolate (strain 18) present on a patient care table was identified as methicillin-resistant S. aureus with intermediate sensitivity to vancomycin (VISA). Our observations suggest the need for containment measures (good antisepsis practices) to avoid the possible transmission of resistant bacterial agents for the veterinary hospital environment.

Description: Global control and elimination of tuberculosis are hindered by the high prevalence of drug-resistant strains, making the development of new drugs to fight tuberculosis a public health priority. In this study, we evaluated 118 extracts from 58 Venezuelan plant species for their ability to inhibit the growth of Mycobacterium tuberculosis mc26020, using the agar dilution method. Additionally, we determined the ability of these extracts to inhibit the activity of PknB protein, an essential M. tuberculosis serine/threonine kinase, using a high-throughput luminescent assay. Of the 118 extracts tested, 14 inhibited bacterial growth with a minimum inhibitory concentration ≤500 μg/ml, and 36 inhibited the kinase activity with a half-maximal inhibitory concentration

Description: Eye infections caused by bacteria are a serious public health problem among pediatric patients. These diseases, if not properly treated, can cause blindness and impaired vision. The study aimed to evaluate the antimicrobial resistance profiles of the main pathogens involved in eye infections. This study involved pediatric patients enrolled at the “Luigi Vanvitelli” University Hospital of Campania in Naples, Italy, between 2017 and 2019. Of a total of 228 pediatric patients, 73 (32%) tested positive for bacterial infection. In terms of strain distribution, 85% were Gram-positive bacteria, while 15% were Gram-negative bacteria. The most frequently isolated strains were coagulase-negative Staphylococci (60.4%), followed by Staphylococcus aureus (16.4%). The isolated bacteria showed a significant percentage of resistance to multiple antibiotics. Therefore, the identification of the causal bacteria and antimicrobial sensitivity tests are mandatory to select the effective drug for the treatment of eye infections and prevent the development of antibiotic-resistant bacteria.

Description: Fish bacterial pathogens cause diseases which result in a considerable economic impact on the aquaculture industry, necessitating the use of antimicrobials for their control. However, intensive and indiscriminate use of antimicrobials has led to increased occurrence of drug resistance in pathogenic bacteria, as well as normal flora. The aim of the current study was to determine the susceptibility patterns of bacteria isolated from fish, with respect to some commonly used antibiotics and disinfectants. Bacteria were isolated between December 2017 and April 2018 from farmed Nile tilapia, African catfish, goldfish, and koi carp in Kirinyaga County, Kenya. Antibiotic and disinfectant susceptibility patterns of 48 isolates belonging to the genera Aeromonas, Proteus, Klebsiella, Citrobacter, Salmonella, Streptococcus, Pseudomonas, Escherichia, Serratia, and Micrococcus were established using the Kirby–Bauer disc diffusion method and agar well diffusion technique, respectively. The antibiotics evaluated included ampicillin, tetracycline, co-trimoxazole, streptomycin, kanamycin, gentamicin, co-trimoxazole, and chloramphenicol, while the disinfectants tested were quaternary ammonium compound, formalin, hydrogen peroxide, sodium hypochlorite, and iodine. All the bacteria except Micrococcus, Escherichia, and Salmonella species showed multiple drug resistance patterns. Streptococcus showed resistance to six antibiotics, while Proteus, Pseudomonas, and Serratia were resistant to five antibiotics. The multiple antibiotic resistance index ranged from 0.1 to 0.8, with Streptococcus spp. having the highest score value. All the organisms were sensitive to gentamicin, while co-trimoxazole and ampicillin showed the highest resistance at 73% (n = 34) and 62% (n = 31), respectively. Most of the disinfectants showed antibacterial activity with varying magnitudes. The isolates were 100% sensitive to hydrogen peroxide and formalin, but were resistant to sodium hypochlorite at recommended user-dilution. The study has shown that some of the bacterial isolates were resistant to common antibiotics and disinfectants; thus, it is recommended to include an antibiogram whenever making any therapeutic decision. The resistant bacteria may transmit resistance genes to other fish bacteria and also to human bacteria, thus making it difficult to treat the resultant disease(s); thus, there is a possibility that these resistant bacteria may be transmitted to humans who consume or handle the carrier fish. It is, therefore, advisable that fish are cooked properly before consumption, so as to kill bacteria that may be present.

Description: Biotic and abiotic factors cause an enormous amount of yield and economical loss. However, endophytes can play a significant role in enhancing the tolerance of plants. Endophytes systematically colonize different parts of the host, but plants use a variety of defense mechanisms towards microbial infection. However, they have to survive the oxidative environments, and endophytes like Enterobacter sp. encode superoxide dismutases, catalases, and hydroperoxide reductases to cope up the oxidative stress during colonization. On the contrary, others produce subtilomycin which binds with flagella to affect flg22-induced plant defense. The behavior of endophytes can be affected by different genes in hydrolase activity when they come into contact with the host plant. The lifestyle of endophytes is influenced by environmental factors, the host, and microbial genotypes, as well as an imbalance in nutrient exchange between the microbe and the host. For instance, induction of PiAMT1 in root endophyte Piriformospora indica indicates depletion of nitrogen which plays as a triggering factor for activation of the saprotrophic program. Microbes enhance disease resistance through induced systemic resistance (ISR), and Bacillus cereus triggers ISR against Botrytis cinerea through an accumulation of the PR1 protein and activates MAPK signaling and WRKY53 gene expression by the JA/ET signaling pathway. Similarly, Trichoderma arundinaceum produces trichodiene that affects Botrytis cinerea through induction of defense-related genes encoding salicylic acid (SA) and jasmonate (JA). Overall, endophytes can play a vital role in disease management.

Description: Soil stabilization is a mechanical or chemical alteration of one or more soil properties to create an improved soil material possessing the desired engineering properties. The aim of this article was to review bioenzyme-based soil stabilization techniques with an emphasis on bioenzymes production, mechanism of soil stabilization and future challenges, and opportunities of the sector. Soils are stabilized to increase strength and durability or to prevent erosion and dust generation. Cost-effective soil stabilization technology has been a fundamental part of any construction and is very important for economic growth in any country. In some cases, construction has been challenged due to the high cost of soil stabilization processes. Besides, methods of stabilizations using common stabilizing agents are getting costly. Currently, there is a growing interest to identify new and green technology to improve construction techniques and to expand the road network. Therefore, the search for new materials and improved techniques to process the local materials has received an increased focus. For developing countries, bioenzymes are now creating an opportunity to improve soil stability with tremendous effectiveness in the overall process of soil stabilization. In the world, bioenzymes have been used in different projects for several years and are generally proprietary products, often of patented formulation that needs intensive field tests. Currently, the use and production of bioenzymes is becoming the most promising key for the advancement of a country by saving time, energy, and finance. It also reduces environmental pollution due to carbon emission by the conventional stabilizers. Thus, a better understanding of this emerging technology is of utmost importance to exploit any improvement it can offer to soil stability. With little research and practice, it is possible to produce soil stabilizing bioenzymes using local raw materials. Due to this, production of low cost, easily and widely applicable, and environmentally friendly enzymatic formulations from locally available raw materials should be the interest of research and academic institutes of any country.

Description: Identifying Bacillus cereus with conventional methods is neither specific nor rapid because of the close relatedness of the B. cereus group, hence the need for molecular methods. Genotypic profiling of B. cereus isolates from food was obtained by Random Amplified Polymorphic DNA-polymerase chain reaction (RAPD-PCR) using OPR13 primer. A dendrogram was drawn with the Numerical Taxonomy System of Statistic (NTSYS) software. Thirty of the isolates were subjected to molecular identification by 16S rDNA sequencing. The thirty sequences were deposited in GenBank for accession number. Phylogenetic relationship of the 16S rDNA sequence obtained was carried out with the Multiple Alignment using Fast Fourier Transform (MAFFT) software version 7.0. The evolutionary tree was drawn using the Molecular Evolutionary Genetics Analysis (MEGA 6) software. The dendrogram generated for the RAPD profile showed that all the strains are closely related, with a similarity coefficient of 70%. The isolates were confirmed with 16S rDNA sequencing as B. cereus. The thirty sequences deposited in GenBank were given accession numbers: KX574760–KX574769, KX610811–KX610820, MT757957-MT757963, and MT772282-MT772284. By comparing the phylogenetic relationship, eleven of the strains did not cluster with the reference strains from the GenBank but form distinct clades, which means they are likely to be of different ancestors. Conventional methods rarely differentiate bacteria of the same species into clade, neither can it describe their ancestral lineage. Therefore, it is important to employ molecular methods in identifying bacteria to give detailed information about them.

Description: Background. The occurrence of urinary tract infection in presence of urolithiasis is frequently noted; however, microbial agents of urolithiasis and their antimicrobial susceptibility patterns remain underinvestigated. This study aimed to identify the microorganisms isolated from urine and stone matrices to determine their antimicrobial susceptibility, to find the association between the pathogens of urine and stone matrices, and to perform the biochemical analysis of stones. Methods. A total of 88 cases of urolithiasis admitted for elective stone removal at Department of surgery, B.P. Koirala Institute of Health Sciences (BPKIHS), were enrolled. Preoperative urine culture and postoperative stone culture were performed. Isolation, identification, and AST were done by the standard microbiological technique. Further qualitative biochemical analysis of stones was also attempted. Result. Among 88 stone formers recruited, culture of urine, whole stone, and nidus yielded the growth of bacteria 44, 32, and 30, respectively. Bacteria isolated from urine culture correlated with those from stone matrices with a sensitivity of 90%, specificity of 79.69%, PPV of 63.64%, and NPV of 95.45%. Escherichia coli (46.7%) was the most common bacteria followed by Klebsiella pneumoniae (16.7%) and Proteus mirabilis (13.3%) from urine and stone cultures. Almost all the uropathogens isolated were susceptible to commonly used antibiotics. Calcium oxalate (84.1%) was common biochemical constituent found in stone formers followed by calcium oxalate + phosphate (8%). Conclusions. The association of microorganism isolated from urine and nidus culture was significant that can predict the source of infective stone; however, in some cases, microorganisms and the antimicrobial susceptibility pattern from urine and nidus were different. This study emphasizes the use of appropriate antimicrobial agents to prevent the regrowth of residual stones and minimize the risk of infectious complications after surgical removal of stones.

Description: Introduction. The burden of bloodstream infections (BSIs) has been warranted in Ethiopia. Globally, the emergency and raised resistance rate of bacterial antimicrobial resistance is becoming a prominent problem, and it is difficult to treat patients having sepsis. In this review, we aimed to determine the pooled prevalence of bacterial isolates among presumptive patients with bloodstream infections in Ethiopia. Methods. A systematic search was performed from PubMed/MEDLINE, Scopus, HINARI, ScienceDirect, and Google Scholar electronic databases using PRISMA guidelines. The data analysis was carried out using STATATM version 14 after the records were cleaned and sorted out. Results. A total of 26 studies with 8,958 blood specimens and 2,382 culture-positive bacterial isolates were included for systematic review and meta-analysis. The meta-analysis derived a pooled culture-positive bacterial prevalence which was 25.78% (95% CI: 21.55–30.01%). The estimated pooled prevalence of Gram-positive and Gram-negative bacterial isolates was 15.50% (95% CI: 12.84–18.15%) and 10.48 % (95% CI: 8.32–12.63%), respectively. The two common Gram-positive bacteria isolated from patients suspected of BSIs were coagulase-negative Staphylococcus with a pooled prevalence of 5.75% (95% CI: 4.58–6.92%) and S. aureus 7.04 % (95% CI: 5.37–8.72%). Similarly, the common Gram-negative bacterial isolates and their estimated pooled prevalence were E. coli 1.69% (95% CI: 1.21–2.16%), Klebsiella species 7.04 % (95% CI: 5.37–8.72%), Pseudomonas species 0.39% (95% CI: 0.08–0.70%), Salmonella species 1.09% (95% CI: 0.79–1.38%), and Streptococcus pyogenes 0.88% (95% CI: 0.54–1.22%). Conclusion. The prevalence of bacterial isolates among presumptive patients suspected to BSIs in Ethiopia remains high. Furthermore, we found a remarkable variation in the pathogen distribution across the study setting.

Description: Viral infections are causing serious problems in human population worldwide. The recent outbreak of coronavirus disease 2019 caused by SARS-CoV-2 is a perfect example how viral infection could pose a great threat to global public health and economic sectors. Therefore, the first step in combating viral pathogens is to get a timely and accurate diagnosis. Early and accurate detection of the viral presence in patient sample is crucial for appropriate treatment, control, and prevention of epidemics. Here, we summarize some of the molecular and immunological diagnostic approaches available for the detection of viral infections of humans. Molecular diagnostic techniques provide rapid viral detection in patient sample. They are also relatively inexpensive and highly sensitive and specific diagnostic methods. Immunological-based techniques have been extensively utilized for the detection and epidemiological studies of human viral infections. They can detect antiviral antibodies or viral antigens in clinical samples. There are several commercially available molecular and immunological diagnostic kits that facilitate the use of these methods in the majority of clinical laboratories worldwide. In developing countries including Ethiopia where most of viral infections are endemic, exposure to improved or new methods is highly limited as these methods are very costly to use and also require technical skills. Since researchers and clinicians in all corners of the globe are working hard, it is hoped that in the near future, they will develop good quality tests that can be accessible in low-income countries.

Description: The emergence and persistence of antibiotic resistance remain formidable health challenges. This study aimed at detecting and profiling antibiotic resistance of bacterial contaminants in vended food and the environment. Seventy antibiotic-resistant bacterial isolates were isolated from fried fish, African sausages, roasted meat, smokies, samosa, chips (potato fries), vegetable salads, and soil samples collected from Embu Town and Kangaru Market in Embu County, Kenya. The antibiotic susceptibility test, morphological and biochemical characterization, antibiosis assay, polymerase chain reaction-based detection of antibiotic resistance genes, and sequencing of the 16S rRNA gene were done. Analysis of variance on all measured data was done, and Tukey’s honest test was used to compare and separate mean diameters of zones inhibition. Resistance of bacterial isolates to antibiotics was chloramphenicol (90%), cefotaxime (84.29%), nalidixic acid (81.43%), tetracycline (77.14%), amoxicillin (72.86%), gentamycin (48.57%), streptomycin (32.86%), and trimethoprim + sulphamethoxazole (30%). Isolate KMP337, Salmonella spp., exhibited highly significant antibiosis against S. aureus recording a mean inhibition diameter and standard error (SE) of 16.33 ± 0.88 mm, respectively, at P=0.001. The 70 bacterial isolates belonged to Bacillus, Paraclostridium, Lysinibacillus, Virgibacillus, and Serratia genera. The study isolated Bacillus wiedmannii (KC75) which is a risk group 2 as well as Serratia marcescens (KMP95) and Bacillus anthracis (KS606) which are risk group 3 organisms. The presence of antibiotic resistance genes Tet A, BlaTEM, StrB, Dfr A, Amp, and FloR genes was confirmed by a polymerase chain reaction. Samples from Kangaru Market recorded a higher (88.57%) proportion of resistant isolates as compared to isolates from Embu Town (11.43%). The study confirmed the presence of antibiotic-resistant bacteria in vended fast food and the soil in Embu Town and Kangaru Market. This study calls for continuous monitoring of bacterial status and hygienic handling of vended food.

Description: Diarrhea remains as a high health burden, especially to children in low-income countries including Ethiopia. Diarrheagenic Escherichia coli have been commonly associated as bacterial pathogens causing diarrheal disease among children. This systemic review and meta-analysis was intended to determine the pooled prevalence of Escherichia coli in under-five children with diarrhea in Ethiopia. A comprehensive search in PubMed, Google Scholar, ScienceDirect, ResearchGate, and Google search engine and manual searching were done for this systematic review and meta-analysis. The eligibility criteria for selecting studies were studies involving under-five children with diarrhea in Ethiopia, published articles, cross-sectional studies, and articles reported in English. The study was conducted based on the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) checklist. The data analysis was done using STATA 16.0 software. Cochran’s Q-test and I2 statistics were used for the assessment of heterogeneity. The random-effect model was used to estimate the pooled prevalence of Escherichia coli. A total of 797 articles were initially retrieved, and finally, 11 studies met the eligibility criteria and were included in the final meta-analysis. The pooled prevalence of Escherichia coli was 25% (95% CI: 9, 41). The pooled prevalence was varied by region, detection method, and sample size. The high prevalence emphasizes that Escherichia coli is a potential pathogen in under-five children with diarrhea in Ethiopia.

Description: Pathogenesis of Aspergillus flavus on important agricultural products is a key concern on human health due to the synthesis and secretion of the hazardous secondary metabolite, aflatoxin. This study identified and further characterized aflatoxigenic A. flavus from groundnuts sampled from sundry shops in Kenya using integrated morphological and molecular approaches. The groundnuts were plated on potato dextrose agar for isolation and morphological observation of A. flavus based on macroscopic and microscopic features. Molecular characterization was done through amplification and comparison of the partial sequence of the ITS1-5.8S-ITS2 region. The expression analysis of aflR, aflS, aflD, aflP, and aflQ genes in the aflatoxin biosynthesis pathways was conducted to confirm the positive identification of A. flavus. The gene expression also aided to delineate toxigenic isolates of A. flavus from atoxigenic ones. Morphologically, 18 isolates suspected to be A. flavus were identified. Out of these, 14 isolates successfully amplified the 500 bp ITS region of A. flavus or Aspergillus oryzae, while 4 isolates were not amplified. All the remaining 14 isolates expressed at least one of the aflatoxigenic genes but only 5 had all the genes expressed. Partial sequencing revealed that isolates 5, 11, 12, 13, and 15 had 99.2%, 97.6%, 98.4%, 97.5%, and 100% homology, respectively, to the A. flavus isolate LUOHE, ITS-5.8S-ITS2, obtained from the NCBI database. The five isolates were accurate identification of atoxigenic A. flavus. Precise identification of toxigenic strains of A. flavus will be useful in establishing control strategies of the fungus in food products.

Description: Background. Neonatal sepsis diagnosis is a challenge because of its nonspecific presentation together with low sensitivity of the time-consuming bacterial cultures. So, many sepsis markers, like C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6), are emerging to improve its diagnosis. Aim. This study was done to investigate the role of CRP, PCT, and IL-6 in promoting the early diagnosis of neonatal sepsis in an attempt to decrease morbidity and mortality. Methods. This cross-sectional study was conducted on 50 neonates suspected with sepsis enrolled from the neonatal intensive care unit (NICU) of Zagazig University Hospitals, Egypt. Blood cultures for these neonates were done before starting antibiotics. Also, bacterial DNA was revealed from the blood by broad-range 16S rDNA polymerase chain reaction (PCR). Measurements of CRP using the immunoturbidimetry method, PCT using fluorescence immunoassay quantitative method, and IL-6 using commercially available ELISA kit were done to all enrolled neonates. Results. Forty-one neonates with proved sepsis were found to be positive in blood culture and/or PCR for bacterial 16S rDNA. The most common isolated organisms were Klebsiella (61.3%), followed by E. coli (9.7%) and CONS (9.7%). We detected much significant higher levels of PCT, CRP, and IL-6 in the proved sepsis group than the suspected neonatal sepsis cases (p≤0.001, 0.001, and 0.004, respectively). Serum PCT levels showed the highest sensitivity, specificity, PPV, NPV, and accuracy of 97.6%, 89%, 97%, 88.9%, and 96% than other studied sepsis markers. Conclusion. PCT has satisfactory characteristics as a good marker than IL-6 and CRP for the diagnosis of neonatal sepsis.

Description: Inadequate use of antibiotics has led to spread of microorganisms resistant to effective antimicrobial compounds for humans and animals. This study was aimed to isolate cultivable strains of actinobacteria associated with Baikal endemic alga Draparnaldioides baicalensis and estimate their antibiotic properties. During this study, we isolated both widespread and dominant strains related to the genus Streptomyces and representatives of the genera Saccharopolyspora, Nonomuraea, Rhodococcus, and Micromonospora. For the first time, actinobacteria belonging to the genera Nonomuraea and Saccharopolyspora were isolated from Baikal ecosystem. Also, it was the first time when actinobacteria of the genus Nonomuraea were isolated from freshwater algae. Some rare strains demonstrated activity inhibiting growth of bacteria and yeasts. Also, it has been shown that the strains associated with Baikal alga D. baicalensis are active against both Gram-positive and Gram-negative bacteria. According to this study and previously published materials, diversity of cultivable actinobacteria and rare strains isolated from D. baicalensis is comparable to that of cultivable actinobacteria previously isolated from plant sources of Lake Baikal. Also, it exceeds the cultivable actinobacteria diversity previously described for macroinvertebrates, water, or sediments of Lake Baikal. The large number of rare and active strains associated with the endemic alga D. baicalensis could be the promising sources for biopharmaceutical and biotechnological developments and discovery of new natural compounds.

Description: Background. Carbapenem resistance among Gram-negative isolates caused by the production of the metallo-β-lactamase (MBL) enzyme is being increasingly reported worldwide. One of the newly emerged metallo-β-lactamases is New Delhi metallo-β-lactamase. Data regarding its occurrence in hospital setting and percentage prevalence among different Gram-negative bacterial isolates are lacking in our part. This study has been undertaken for determining the presence of the bla NDM-1 gene among the clinical isolates of imipenem-resistant Gram-negative bacteria in a tertiary care center in Dharan, Nepal. Methods. A total of 75 imipenem-resistant Gram-negative isolates were studied. These were screened for metallo-β-lactamase (MBL) production by phenotypic assays such as double-disc synergy test (DDST) and combined disc diffusion test (CDDT). PCR was performed for the molecular detection of gene NDM-1. Ten-disc method was performed to detect the presence of ESBL, AmpC, carbapenamase, and K1 β-lactamase production. Results. Using the molecular method, bla NDM-1 was detected in 36% of the isolates. Phenotypically, double-disc synergy test (DDST) and combined disc diffusion test (CDST) detected MBL production in 38.7% and 37.3% of the isolates, respectively. Ten-disc method detected ESBL in 26.6% of the isolates, but none of the isolates was found to be AmpC, carbapenamase, and K1 β-lactamase producers. Conclusion. A high percentage of the NDM-1 producer was noted among imipenem-resistant GNB. Apart from performing only antimicrobial sensitivity test, phenotypic and molecular screening should be employed to find out the actual number of metallo-β-lactamase producers and the existence of the resistance gene.