API Kit Research Articles

Investigation of the Most Common Specimens in the Pediatric Hospital in Kirkuk and the Epidemiology of Predominant Bacteria Using Multiple Antibiotic Resistance Indices

Background: Urinary tract infections (UTIs) are among the most prevalent infectious diseases in children and are a leading cause of antibiotic use and pediatric hospitalization. Objectives: This study aimed to investigate the most common specimens and determine the frequency of Multiple Antibiotic Resistance Indexes (MARI) for predominant bacteria in the pediatric hospital of Kirkuk. Methods: A total of 299 different samples were collected from the pediatric hospital in Kirkuk province between May 1st and October 1st. The samples were cultured on blood agar, mannitol agar, and MacConkey agar and incubated for 24 hours at 37°C. Identification was based on morphological and microscopic examination, as well as the API kit. Results: The results showed that most of the samples were urine, with 221 samples collected. Of these, 66 (28.50%) showed positive growth, and 155 (71.49%) showed negative growth. Blood samples were the second most common, with 51 samples collected, of which 4 (9.61%) were positive, and 47 (90.38%) were negative. Stool samples amounted to 13, with 2 (15.38%) showing positive growth, and 11 (84.61%) showing negative growth. Cerebrospinal fluid (CSF), throat swab, and vaginal swab samples amounted to 11, 2, and 1, respectively, with no bacterial growth observed in them. The majority of isolates were from urine, with Escherichia coli being the most common species (28 isolates, 42.42%). Other species isolated included Klebsiella pneumoniae (13 isolates, 19.69%), Staphylococcus spp. (12 isolates, 19.05%), Enterobacter spp. (7 isolates, 10.61%), and Pseudomonas spp. (4 isolates, 6.06%). Proteus spp. and Streptococcus spp. each accounted for 1 isolate (1.25%). The bacteria isolated from blood were Acinetobacter spp., Enterobacter spp., Pseudomonas spp., and Klebsiella spp., each representing 1 isolate (1.25%) of the total. Two (100%) E. coli isolates were obtained from stool samples. E. coli isolates from urine were resistant to ampicillin (Am), tetracycline (Te), amoxicillin/clavulanic acid (AMC), and doxycycline (Do). Klebsiella spp. were resistant to Am, Staphylococcus spp. to erythromycin (E) and azithromycin (AZM), Enterobacter spp. to amikacin (AK) and AMC, and Pseudomonas aeruginosa to gentamicin (CN), all exhibiting 100% resistance. The MARI was greater than 0.2 in 27 isolates of E. coli (96.42%), 13 isolates of Klebsiella spp. (100%), 11 isolates of Staphylococcus spp. (91.66%), 7 isolates of Enterobacter spp. (100%), and 4 isolates of P. aeruginosa (100%). Conclusions: The current study concludes that urinary tract infection is the most common pathological condition among the young age group. Urine is one of the most frequently examined samples, with E. coli being the most predominant pathogen. The study also highlights the prevalence of antibiotic-resistant bacteria, as evidenced by the elevated MARI in isolates of E. coli, K. pneumoniae, Enterobacter spp., and Pseudomonas spp.

Isolation, Selection, and Biological Evaluation for Bacteria that Fix Nitrogen and Produce Indole-3-Acetic Acid from Paddy Soils in Vietnam

Introduction/Background Acidic soils are limiting the production of crops and indirectly harming the environment due to the use of nitrogen (N) chemical fertilizer. Therefore, the current study aims to isolate a promising N2-fixing candidate to solve this issue in some communes of Phu Luong District, Thai Nguyen Province, Vietnam. Materials and Methods The bacteria were isolated using the Ashby medium. The IAA generation and nitrogen fixation were assessed using spectroscopy. The selected bacteria were tested at temperatures ranging from 28°C to 42°C and pH levels of 4.0 to 9.0. As a result, the Box-Behnken model yielded a growth equation. The API kit test was used to measure the biochemical properties of the selected bacteria. Results The result demonstrated that there were ten nitrogen-fixing bacteria (NFB) isolates with capacities of N2 fixation and IAA production at 2.35–24.61 and 0.00–119.13 µg/ml. Among them, the NL3 strain was the best strain. The optimum condition to grow the NL3 strain was calculated as 5.44 days at pH 7.10 and temperature of 32.18°C to reach a bacterial density of 9.77929×108 CFU/ml. In addition, the NL3 strain was identified as an Azotobacter species. The genomic analysis revealed that the genome of Azotobacter sp. NL3 was nearly 5.4 Mb long and contained 45 N2 fixation-relating genes and 7 IAA production-relating genes. Conclusion Not only can this study provide insight into the features of Azotobacter spp., but it also introduces a potent candidate that can improve soil health and crop yield by fixing N2 and producing IAA in order to limit the use of N chemical fertilizer for a sustainable agriculture. Therefore, the selected bacterial strain should be further tested under the local field conditions and subsequently commercialized as a biofertilizer.

Identification of Vibrio Cholerae using Analytical Profile Index (API 20E)

Cholera outbreak have resulted in the deaths of millions of people globally. The cholera-causing Vibrio cholerae, has multiple serogroups, and produces epidemics when two strains, O1 and O139, are present. With the exception of recent, rare instances, V. cholerae O139 caused prior outbreaks; however, V. cholerae O1 was responsible for all following epidemics. Hence, identification of the V. cholerae is an important step in the control of cholera outbreak especially in the local community where inadequate facilities for prognosis limit prompt treatment of patients. A total of 15 isolates previously identified with molecular method were subjected to TCBS Agar and Analytical Profile Index. All the sucrose-fermenting colonies on TCBS agar were correctly identified with API kit. The study concluded that API kit can be substituted for molecular method in prognosis of Cholera outbreak especially in resource-limited Countries due cost implication.

Biofilm-Forming Antibiotic-Resistant Bacteria in Water From Distribution Systems: Occurrence and Public Health Implications.

Biofilm is a structurally-connected microbial community, covered by a self-produced polymeric matrix and adhered to biotic or abiotic surfaces. This study aimed to evaluate the occurrence of biofilm-producing antibiotic-resistant bacteria in water from distribution systems. Water samples were taken from 32 tanks across Ondo City and Akure metropolis, Nigeria. Information regarding the sanitation status of the tanks was gathered by observation and oral interviews. The physicochemical properties were determined using standard methods. Using the pour plate technique. Agars included serially diluted water samples were inoculated onto plate count agar, mannitol salt agar, Salmonella-Shigella agar, MacConkey agar, and cetrimide nutrient agar to assess total viable bacteria, Staphylococcus aureus, Salmonella and Shigella, coliforms, and Pseudomonas aeruginosa, respectively. Eosin-methylene blue agar was used to cultivate Escherichia coli and Enterobacter aerogenes. Pure isolates were characterised using API kits and assessed for antibiotic resistance and biofilm production employing the Kirby-Bauer and tissue culture plate techniques, respectively. The ages of the water tanks ranged from 1 to 25 years old; all tanks had cover-lids; 13 (40.63%) had water guards while 12 (37.5%) underwent water treatment. The physicochemical properties chiefly fell within WHO standards for drinking water. One hundred and eighty-seven isolates were obtained. S. aureus (15.51%) had the highest frequency while Salmonella enterica (3.2%) had the lowest frequency. Thirty-six percent of the isolates were strong biofilm producers, while 20.67% Gram-negative and 18.69% Gram-positive bacterial isolates were antibiotic-resistant. This study revealed a high occurrence of biofilm-forming bacteria and prevalence of antibiotic-resistant bacteria in water distribution systems, emphasizing the urgency of improving water quality for public health protection.

COMPARATIVE IDENTIFICATION APPROACH FOR GUM-PRODUCING LACTOBACILLI IN PALM WINE

Introduction: Species of lactobacilli are responsible for the consistency and soluble white colouration of palm wine through the production of gum. Phenotypic and physiological characteristics as basis for identification of lactobacilli is often with low level of discrimination. Objection: This study aimed at comparative identification approach of gum-producing lactobacilli using amplified ribosomal DNA restriction analysis (ARDRA) and API kit 50 CHL. Methodology: Four hundred colonies of lactobacilli isolated from seventy-two fresh palm wine samples and two hundred were used for genomic DNA extraction, 16S rDNA analysis and digestion of amplicons with two endonucleases: Hae III and Bash 12361. Results: Twenty ARDRA pattern group were generated and representatives of each were randomly chosen for sequence and API kit analysis. The size of the amplified products (̴ 1500 bp), digested amplicons (100 - 1000 bp) and the ARDRA profiles; A-T. ARDRA screening revealed unique patterns among the two hundred isolates, with the same pattern for some of the isolates. The 16S rDNA gene sequence analysis showed that the isolates were divide into 20 groups of three genera: Lactobacillus (65%), Leuconostoc (30%), and Lactococcus (5%) whereas API kit identified two genera; Lactobacillus (75%) and Leuconostoc (25%). Gene sequence similarity of all the representative isolate for 16S rDNA (99 - 100%) while API kit (74.8 – 99.9%) with 55% overall similarity in bacterial identified. ARDRA showed higher similarities in identification and discrimination of bacterial species or between closely related groups isolated from palm wine compared to API kit. Conclusion: ARDRA is a good tool for lactobacilli identification based on the results from this study.

Two Years Study of Frequency and Antimicrobial Resistance Patterns of Escherichia coli Strains Isolated from Clinical Specimens in Northern Iran

Background: According to the reports from the World Health Organization, increased use of antibiotics and bacterial resistance has become a worldwide issue. Resistance to antimicrobial agents in Escherichia coli clinical strains is increasing. Objectives: The objective of this study was to determine the antibiotic resistance patterns and frequency of multidrug resistance (MDR) phenotype in E. coli isolated from patients in two major hospitals in Sari, north Iran. Methods: In this descriptive-analytical study, a total of 13322 clinical specimens were collected from patients. All the specimens were evaluated to determine the presence of E. coli strains using conventional biochemical tests and API kit. Susceptibility testing against twelve antibiotics was determined using the disk-diffusion method. Results were interpreted in accordance with the Clinical and Laboratory Standard Institute (CLSI) protocol. Results: Out of 13322 studied samples, 964 (7.23%) E. coli strains were identified. In two hospitals, high resistance to ampicillin and cefalexin was presented in 621 (64.4%) and 402 (41.7%) isolates, respectively. The highest antibiotic resistance was observed in the burn unit, the burn intensive care unit (ICU) and the burn restoration section, while all the strains (eight) that were isolated from the neonatal-ICU, were sensitive to all the tested antibiotics except cefalexin, nitrofurantoin, nalidixic acid, and ampicillin. Also, strains isolated from urine, wound, stool, and blood samples were resistant to all tested antibiotics. Conclusion: Increased resistance to different antibiotics in burn hospitals has created increasing concern. Very high resistance to some antibiotics indicates that these drugs are misused in therapeutic centers and highlights that infection control measures should be arranged in the ICUs of our hospitals.

Prevalence and Characteristics of Yersinia Enterocolitica and Yersinia Pseudotuberculosis from Raw Milk Supplied in Tehran

Background: Foodborne diseases are caused by eating contaminated food. Yersinia species are among the bacteria involved in food contamination, such as meat, poultry, vegetables, and milk. This study aims to investigate the prevalence and characteristics of Yersinia enterocolitica and Yersinia pseudotuberculosis from raw milk in Tehran. Methods: In this study, 360 samples of raw milk were collected from farms around Tehran and examined according to the FDA method. Then, 25 ml of milk samples were added to 225 ml of Peptone Sorbitol Bile Broth and kept for 10 days at 4 °C for enrichment. After 10 days, 0.5 ml of the sample was added to 0.5 ml of 0.72% KOH solution and 0.54% NaCl. After 30 sec, it was cultured in the selected Cefsulodin-Irgasan-Novobiocin agar (CINagar) medium to remove the normal flora. The plates were stored for 48 h at 28 °C. The suspected Bull's eye colonies were purified and phenotyping tests were carried out on the selected colony. The 20 E API kits were used for confirming identification. Results: From 360 raw milk samples, 4 Yersinia isolates (1.1%) including one Y. pseudotuberculosis (0.27%) and three Y. enterocolitica (0.83%) were isolated. In addition to Yersinia, other bacteria such as Klebsiella, Serratia, Citrobacter, and Providencia were isolated from milk samples. Conclusion: The findings showed that clean tap water and healthy cattle in livestock can be effective in preventing Yersinia contamination. Lack of personal and environmental hygiene could cause food contamination by Yersinia and other intestinal bacteria leading to gastrointestinal infections.

The bacterial flora of Oreochromis niloticus and Clarias gariepinus from earthen ponds in Sagana and Masinga, Kenya

Abstract. Karimi RD, Ngeranwa JJN, Njagi ENM, Kariuki S. 2022. The bacterial flora of Oreochromis niloticus and Clarias gariepinus from earthen ponds in Sagana and Masinga, Kenya. Intl J Bonorowo Wetlands 12: 63-73. Food-borne diseases traced to fish consumption have been reported globally, including in Kenya. The aspect of food quality as far as fish consumption is concerned is underestimated in Kenya though aquaculture has been promoted. The bacterial flora of Tilapia (Oreochromis niloticus Linnaeus, 1758) and Catfish (Clarias gariepinus Burchell, 1822) from Masinga Dam and earthen ponds at Sagana fish farm was determined in this study to determine the anti-microbial response of the pathogenic bacteria. Tilapia fish and Catfish samples were collected from Masinga Dam and Sagana farm in the dry and rainy seasons. The fish were skinned, and gut content was taken for laboratory tests. The water and water sediment samples from these two study sites were also collected. Those samples were processed and cultured in MacConkey agar, and the selective media were subcultured in the colonies and then subjected to morphological examination from cultures. Then, the biochemical tests were carried out using commercially available API kits. The study showed the presence of bacterial species belonging to Enterobacter spp. (n=34), Pseudomonas spp. (n=6), Aeromonas spp. (n=5), Vibrio spp. (n=3) and Acinetobacter spp. (n=2) isolates during the dry season, while bacterial species belonging to Enterobacter spp. (n=31), Pseudomonas spp. (n=6), Aeromonas spp. (n=4) isolates during the dry season. The anti-microbial susceptibility analysis showed that the highest resistance rates were found against Ampicillin (Amp) (61.5% of isolates), Amoxicillin (AmL) (65.9% of isolates), Tetracycline (Te) (31.8% of isolates), and Chloramphenicol (C) (27.5% of isolates) while the lowest was Nalidixic acid (Na), Cefuroxime (Cxm) and Streptomycin (S) at (4.4% of isolates) each. All isolates were sensitive to Gentamycin (Gen), Ciprofloxacin (Cip), and Cefotaxime (CTX). The presence of the above organisms, some potentially pathogenic to humans, indicates that improperly handled, undercooked, or consumed raw fish may cause disease in susceptible individuals. At the same time, some isolates’ anti-microbial resistance indicates that the use of antibiotics in aquaculture to promote growth should be studied further with a view to policy formulation.

Effect of gold nanoparticles on the expression of efflux pump mexA and mexB genes of Pseudomonas aeruginosa strains by Quantitative real-time PCR

Antibiotic-resistant Pseudomonas aeruginosa infections are usually difficult to treat, and there are limited antibiotics for treating them. Increased antibiotic resistance of this bacterium, especially in a multidrug form, has caused many problems for treatment. Nowadays, metal nanoparticles are considered as appropriate alternatives to antibiotics. The objective of the present study was to investigate the effect of gold nanoparticles on the expression of MexB and MexA genes in Pseudomonas aeruginosa isolates.Pseudomonas aeruginosa isolate was identified using biochemical tests and an API kit. The antibiotic sensitivitytest for different antibiotics was performed withthe Kirby-Bauer test according to the CLSI standard. The presence of MexB and MexA genes was assessed by PCR. The effect of gold nanoparticles was investigated by microdilution to evaluate the minimum inhibitory concentration, and the expression of MexB and MexA treated genes was done with silver nanoparticles by the Real-Time PCR method.40 Pseudomonas aeruginosa isolates were detected and identified. These isolates showed significant resistance to various antibiotics. All strains were carriers of MexB and MexA genes, and finally, in the expression of MexB and MexA genes,a significant decrease in the expression of these genes was observed in the samples treated with gold nanoparticles compared to non-treated samples.One of the mechanisms of antibacterial activity of gold nanoparticles is through reducing the expression of mexA and mexB genes and thus reducing the number of active efflux pumps at the cell surface.

Biochemical and molecular identification of Gram-positive isolates with β-hemolysis activity isolated from the nasal swab of pigs during the human meningitis outbreak in Badung Regency, Bali-Indonesia.

Background and Aim:The nasal cavity of a pig serves as an entry point and a habitat for the colonization of commensal microbes and pathogenic bacteria. Based on biochemical and serological tests, Streptococcus b-hemolytic Group C was identified as the Gram-positive bacteria, which resulted in the 1994 outbreak and death of thousands of pigs in Bali. Furthermore, this agent is zoonotic and frequently results in the development of meningitis lesions in the infected pig. Recently, a meningitis outbreak in humans was also reported after the consumption of pig-derived foods at Sibang Kaja, Badung-Bali. This study aimed to identify and characterize Gram-positive β-hemolytic organisms collected from nasal swab of pigs from the outbreak area, as well as to compare API Kit and 16S rRNA gene analysis methods.Materials and Methods:This study commenced with the cultivation of two isolates, Punggul Swab Nasal (PSN) 2 and PSN 19, which were characterized by β-hemolysis activity. These samples were then conventionally and molecularly identified using Kit API 20 Strep and 16S ribosomal RNA (rRNA) gene primers, respectively.Results:Using the Kit API 20 Strep, both isolates were identified as Enterococcus faecium, which was previously classified as Group D Streptococci. Based on the 16S rRNA gene sequencing, PSN 2 and PSN 19 were molecularly confirmed to have 99 and 98.1% similarities with E. faecium (NR042054), respectively. Furthermore, both isolates share the same clade in the phylogenetic tree analysis.Conclusion:Using Kit API 20 Strep and 16S rRNA gene analysis, the PSN 2 and PSN 9 Gram-positive isolates with β-hemolysis activity from pig nasal swabs were identified as E. faecium.

Isolation and phenotypic characterization of Gallibacterium anatis biovar haemolytica from a hen with hemorrhagic ooforitis

Objective: the isolation and phenotypically identification of a Gallibacterium anatis biovar haemolytica strain from a hen with hemorrhagic ooforitis; the antimicrobial susceptibility testing of this isolate. Methods and results: a strain of G. anatis biovar haemolytica, was isolated and phenotypic identified by morphological, cultural and biochemical characters examination, with API 20 E, API 20 NE, API STAPH, API ZYM tests and ABIS online software. The antimicrobial susceptibility of the isolate was performed using the standard disk diffusion method. Conclusions: a strain of G. anatis biovar haemolytica was isolated and phenotypically identified from a hen. From our knowledge, this is the first reporting in Romania of isolation and identification of G. anatis biovar haemolytica. The Gah IDSA 161 strain could be phenotypic identified only by ABIS on line software, Pasteurellaceae Database version, unifying the results of four API kits and other biochemical tests. The isolate showed a multi-drug resistant profile to tetracyclines (tetracycline, oxitetracycline, doxicyclin), floroquinolones (enrofloxacin, ciprofloxacin), ampicillin, trimethoprim, nalidixic acid, clindamycin, and it was susceptible to sulfonamide, sulfomethoxazole/trimethoprim, gentamicin, cephalothin, streptomycin, amoxicilin/clavulanic acid.

Occurrence of selected pathogenic microorganisms in raw and processed eggs of snails of the Cornu genus

IntroductionThis study investigated the eggs of Polish-bred edible snails of the Cornu genus as a food and aimed to determine the presence of microorganisms in them of the Salmonella and Listeria genera and ascertain the number of coagulase-positive staphylococci.Material and MethodsRaw material, semi-finished products, and the final product were collected during the production cycle. Testing for the presence of Salmonella spp. and Listeria spp. and measuring of the pathogenic staphylococci contamination level were carried out in accordance with ISO standards. Commercial biochemical tests were used for species identification of bacteria of the Enterobacteriaceae family and Staphylococcus genus. An API kit and a PCR protocol were utilised for species confirmation of the microorganisms of the Listeria genus.ResultsNeither Salmonella nor coagulase-positive staphylococci were found in any of the studied material. Bacteria of the Listeria genus were found in samples taken at every stage of production; however L. monocytogenes was confirmed in samples of the final product.ConclusionThe absence of Salmonella spp. and Staphylococcus aureus in samples of the final product indicates that the required hygiene standard was maintained in the production process of edible snail eggs. Nevertheless, the presence of L. monocytogenes in eggs of common garden snails may pose a potential risk to consumer health.

Common Features and Intra-Species Variation of Cutibacterium modestum Strains, and Emended Description of the Species.

Cutibacterium modestum is a new species coined in 2020 as the fifth species of genus Cutibacterium, which includes Cutibacterium acnes. The species is predicted as a minor but common member of skin microbiome and includes a group tentatively named as “Propionibacterium humerusii”. The description of the species has been provided only with a single strain. To establish the characteristics of C. modestum and search for possible disease-related subtypes, we investigated the biochemical characteristics of eight live strains and performed in silico comparison of nine genomes. The common features, which included the morphology of Gram-stain positive short rods, the negativity of phenylalanine arylamidase, and several unique MALDI-TOF MS spectral peaks, were considered useful in laboratory identification. Pairwise comparisons of the genomes by in silico DNA–DNA hybridization showed similarity values of 98.1% or larger, which were far higher than the subspecies cutoff of 79–80%. The 16S rRNA gene sequences of thirteen isolates and genomes were identical. Their recA gene sequences were identical except for two strains, HM-510 (HL037PA2) and Marseille-P5998, which showed unique one-nucleotide polymorphisms. The biochemical features using API kits were slightly different among the isolates but far closer than those of the nearest other species, C. acnes and Cutibacterium namnetense. Spectra of MALDI-TOF mass spectrometry showed slight differences in the presence of m/z 10,512 (10 kD chaperonin GroS) and three other peaks, further clustering the eight isolates into three subtypes. These results indicated that these isolates did not separate to form subspecies-level clusters, but subtyping is possible by using recA gene sequences or MALDI-TOF mass spectrometry spectra. Moreover, this work has confirmed that a group “P. humerusii” is included in C. modestum.

Comparative Assessment of Matrix-assisted Laser Desorption Ionization-time of Flight Mass Spectrometry (MALDI-TOF-MS) and Conventional Methods in the Identification of Clinically Relevant Yeasts.

Background: Candida species are generally identified by conventional methods such as germ tube or morphological appearance on cornmeal agar (CMA), biochemical methods using API kits, and molecular biological techniques. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) has revolutionized the identification of fungi reducing the turnaround time of days to minutes.Purpose: To compare the performance of MALDI-TOF MS and conventional methods in the identification of clinically relevant yeasts.Materials and methods: In this study, Candida identifications on CMA are compared with the results obtained on MALDI-TOF MS (Bruker Daltonics, Bremen, Germany). Discrepant results were confirmed by sequencing internal transcribed spacer (ITS) regions of rDNA.Results: A total of 114 clinical Candida species isolated from blood cultures were isolated and identified with conventional methods as well as with the MALDI-TOF-MS system. The agreement between the two test results were analyzed using Inter-rater reliability analysis (Cohen's Kappa) in SPSS Software Version 24 (IBM Corp., Armonk, NY). Overall, there was substantial agreement (Cohen's kappa=0.763) between the two methods. A value between 0.61 and 0.80 is classified under substantial. The most frequently isolated bloodstream Candida species included Candida albicans, C. tropicalis, C. parapsilosis, C.lusitaniae, C. glabrata which were accurately identified by MALDI-TOF-MS. When compared with conventional identification methods, MALDI-TOF-MS results are more reliable and rapid for Candida identification.

Identification of dominant lactic acid bacteria and yeast in rice sourdough produced in New Zealand

This study characterised a commercial New Zealand gluten free (GF) rice sourdough and its starter culture composition. Acidity of the mother sourdough, dough before proofing and dough after proofing was determined during the production of rice sourdough bread, and colour was measured for the baked bread. Yeast and lactic acid bacteria (LAB) were enumerated in the rice sourdough samples and representative colonies characterised using API kits and sequenced by the Internal Transcribed Spacer and 16 S rRNA region. Sourdough LAB isolates were identified as Lactobacillus (L.) papraplantarum DSM 10667 and L. fermentarum CIP 102980 and the yeast isolates as Saccharomyces (S.) cerevisiae CBS 1171. Dough acidity increased significantly (p < 0.05) during fermentation due to the metabolic activities of the sourdough cultures. After baking, the colour of the rice sourdough bread crust was similar to that of unleavened wheat bread (golden brown). The improved colour of the rice sourdough bread crust may be a result of combined use of sourdough technique and optimal baking conditions. The results of this study may allow bakers to improve the overall quality of GF rice sourdough baked bread by selecting suitable fermentation and baking parameters.

Inhibition of Staphylococcus aureus LC554891 by Moringa oleifera Seed Extract either Singly or in Combination with Antibiotics.

Bacterial outbreaks caused by Staphylococcus aureus (S. aureus) are interesting due to the existence of multidrug resistant (MDR) isolates. Therefore, there is a need to develop novel ways to control such MDR S. aureus. In this study, some natural agents such as honey bee (HB), extracts of either Moringa oleifera seeds (MSE), or leaves (MLE) and essential oils of garlic, clove, and moringa were studied for their inhibitory activity against this S. aureus pathogen. About 100 food samples including beef luncheon (n = 25), potato chips (n = 50), and corn flakes (n = 25) were investigated for possible pollution with the S. aureus bacteria. The isolated bacteria suspected to belong S. aureus that grew well onto Baird–Parker agar (Oxoid) and shiny halo zones and positive coagulase reaction were selected and identified by API-Kits; all of them that were approved belong to S. aureus (18 strains). The sensitivity of the obtained 18 S. aureus bacterial strains to 12 antibiotics were evaluated; all of them were resistant to ofloxacin; however, other antibiotics tested showed variable results. Interestingly, the S. aureus No. B3 isolated from beef luncheon was resistant to 10 antibiotics out of 12 ones tested. Multiple antibiotic resistance index (MAR) of this S. aureus strain was about 83.3%. Therefore, its identification was confirmed by sequencing of a 16S rRNA gene which approved a successful biochemical identification carried out by API Kits and such strain was designated S. aureus LC 554891. The genome of such strain appeared to contain mecA gene encoding methicillin resistance; it was found to contain hla, hlb, tsst-1, and finbA that encode α-blood hemolysis, β-blood hemolysis, toxic shock syndrome gene, and fibrinogen-binding protein gene, respectively. In addition, the virulence factors viz. sea; seb; sec encoding enterotoxins were detected in the DNA extracted from S. aureus B3 strain. Aqueous extract of Moringa oleifera seeds (MSE) showed inhibitory activity against S. aureus LC 554891 better than that obtained by tetracycline, essential oils or HB. Minimum inhibitory concentration (MIC) of MSE was 20µg/mL. Instrumental analysis of MSE showed 14 bioactive chemical compounds. Combinations of both MSE and tetracycline showed distinctive inhibitory activity against S. aureus LC 554891 than that obtained by either tetracycline or MSE singly.

Enteric Viruses and Bacterial Species from the Faecal Droppings of Bats in Selected Bat Colonies in Ondo State, Nigeria

Background: Pathogenic viruses and bacteria of bat origin causes a lot of diseases that often leads to epidemics and pandemics. Fruit bats were documented reservoir of many viral infections including COVID-19. This study was designed to detect the presence of some enteric viruses of gastroenteritis as well as bacterial species from Eidolon helvum bat species in Nigeria. Methodology: Twenty-five samples were collected between January and March 2018, from six different locations in Ondo State, Nigeria. The samples were tested for the presence of Rotavirus and Adenovirus antigens using the CerTest Biotech 4th generation Quadruple Enzyme Immunoassay (EIA) kit according to the manufacturers’ specification. The bacterial species were isolated by pour plating and identified using standard microbiological methodsand API kits. Results: Throughout the period of this study, two samples (10%) were tested positive for both Norovirus and Adenovirus in bat fecal samples from Epinmin Akoko, Nigeria, while the isolated bacterial species from bats in Owo included Streptococcus Spp, Micrococcus Spp, Staphylococcus aureus, Enterococcus Spp., Staphylococcus epidermidis, Salmonella typhi, Echerichia coli, Klebsiella oxytoca, Acinetobacter iwoffi, Citrobacter freundii, Serratia marcescens, Enterobacter cloacae, Proteus vulgaris, Enterobacter agglomerans and Proteus mirabillis. Conclusion: The isolation of these microbes are suggestive of infectious agents that can be of threat to public health and caution bush meat consumers from erratic consumption of unverified bush meat culinaries. The data obtained from this study is valuable for public health management as well as disease prevention and control

Isolation and Identification of Bacteria from Phytoremediation Plant of Heliconia psittacorum

Phytoremediation is a technology that uses plants to degrade, assimilate, metabolize, or decompose organic matter and pollutants. Critical criteria for plants used for phytoremediation include economic benefits, harvest management, and side-use. This cost-effective plant-based recovery utilizes exceptional plant and microorganism capabilities to focus on environmental elements and compounds and metabolize various molecules in their tissues to decompose elements such as heavy metals, toxic substances, and organic pollutants are the main targets. This study aims to analyze the concentration of Gram-positive and Gram-negative bacteria and identify the bacteria using biochemical assays and commercialized API Kit (Biomerieux, France). Three plant samples of Heliconia psittacorum were analyzed for their root and soil sample which showed the concentration of Gram-positive bacteria is higher compared to Gram-negative bacteria which ranged from 1.9 x 108 to 3.95 x 109, while gram-negative bacteria ranged from 2.25 x 107 to 2.95 x 109. Based on the identification test, seven bacteria were identified. Four gram-negative bacteria are Enterobacter aerogenes, Escherichia coli, Pseudomonas aeruginosa and Chryseobacterium indologenes which are generally bacteria that are common to the environment. Meanwhile, three gram-positive bacteria include Staphylococcus aureus, Staphylococcus xylosus, and Staphylococcus lentus have a high impact on metal degradation in the effluent. In conclusion, the concentration of bacteria is relatively higher in the root area of the plant which dominated by gram-positive bacteria and the bacteria that were identified have a prominent effect on the phytoremediation process.

Changes of microflora in high pressure treated Indian white prawn (Fenneropenaeus indicus)

ABSTRACTThis study was to investigate the piezotolerance and diversity indices of microflora of Indian white prawn (Fenneropenaeus indicus) after high pressure (HP)-treatment. Indian white prawns subjected to HP-treatments and its effect was studied up to species level and compared with unpressurized samples. The bacterium was identified by using bacterial identification schemes, biochemical tests and API kits (bioMérieux, Marcyl’Etoile, France). Diversity analysis was performed using PRIMER (Plymouth Routines in Multivariate Research) software v 5.2.2. The significant elimination of microflora was found to be proportional with the pressure level. In the case of spore formers, mere destruction was noticed after HP-treatment. Arthrobacter spp., Listeria grayi and Corynebacterium spp. were the most piezotolerant bacteria in HP-treated samples. Diversity indices revealed a significant reduction of microflora of Indian white prawn. The apparent reduction of microflora with pressure level was clearly evident from the diversity indices; moreover a diminished piezotolerance of Gram negative spoilage bacteria was also observed.

Weissella, a novel lactic acid bacteria isolated from wild Sumatran orangutans (Pongo abelii).

Aims:This study aimed to isolate and identify lactic acid bacteria (LAB) in wild Sumatran orangutans to provide more information about LAB diversity derived from Sumatran orangutan feces.Materials and Methods:Fecal sampling from three female orangutans, around 35 years old, was carried out in the wild forest areas at the research station of Suaq Belimbing Gunung Leuser National Park located in the South Aceh district. Orangutan fecal samples were taken in the morning when the orangutans first defecated. The orangutans were above the tree, which is approximately 12-15 m from the ground where feces were found.Results:Fermentation testing using the API 50 CHL Kit showed that OUL4 isolates were identified as Lactococcus lactis ssp. lactis with an identity value of 73.5%. Homology analysis demonstrated that the OUL4 isolates have 93% similarity to Weissella cibaria, and phylogenetic trees constructed using Mega 7.0 also showed that OUL4 isolates are related to W. cibaria.Conclusion:These results show that there is a difference in identification between biochemical testing with API kits and molecular analyses on LAB isolates from wild Sumatran orangutans. Based on 16S rRNA gene homology, the OUL4 LAB isolates from wild Sumatran orangutans have 93% homology to W. cibaria.