Efficacy of leaf extracts of Psidium guajava (L.) on enteric bacterial isolates from faecally impacted groundwater

The efficacy of leaf extracts of guava Psidium guajava (L.) were assessed on enteric bacteria isolated from two wells (W1 and W2) prone to faecal contamination in Akure, Nigeria. The load of enteric bacteria in the water samples were determined using standard microbiological method. Leaf extracts of P. guajava (L.) were prepared using ethanol and hot distilled water and the antibacterial effects of the extracts on bacterial isolates were determined using agar well diffusion method. Phytochemical components of the leaf extracts were carried out using standard methods. Results revealed the following mean concentrations of enteric bacteria in W1 and W2: Salmonella (3.8 and 3.9 log10 CFU/100 ml) and Shigella (3.6 and 3.7 log10 CFU/100 ml). The ethanol leaf extracts of P. guajava (L.) exhibited a zone of inhibition of 22 mm at 200 mg/ml on Shigella and no zone of inhibition was observed on Salmonella. The phytochemical components of ethanol and hot distilled water leaf extracts of P. guajava (L.) revealed the presence of saponin as (13.64 mg/g) and (59.82 mg/g). The findings of this study revealed that ethanol leaf extracts of P. guajava (L.) may be useful as antibacterial agent against Shigella.

Value of polymerase chain reaction in etiological diagnostic of infectiove endocarditis

Introduction Identification of a causative agent in patients with infective endocarditis (IE) is crucial for diagnostic and prescribing etiotropic therapy which defines positive outcome of a disease. High rate of a culture negative IE and inaccurate results of traditional microbiological methods raise a concern. So the methods of etiological diagnostics in IE are in need of development, particularly introduction of polymerase chain reaction (PCR) method might be helpful. Aim Modernisation of the algorithm of IE etiological diagnostic by introducing PCR Materials and methods The study included 85 cases of IE [first episode of IE (n=79), recurrence/relapse (n=6)] verified by DUKE criteria 2009, 2015, hospitalized in Moscow primary hospital from 2012 to 2017. All patient had venous blood investigated both with microbiological method and with broadrange and specific PCR. Following microorganisms' DNA were assessed by PCR: Staphylococcus spp. (MRCoNS, S. aureus and others), Streptococcus spp. (S. agalactiae, S. pyogenes and others), Enterococcus spp. (E. faecium, E. faecalis and others), Enterobacteriaceae, Klebsiella spp. (K. pneumoniae and others), E. coli, Proteus spp., A. baumanii, P. aeruginosa, Fungi (C. albicans, C. glabrata, Aspergillus spp. and others). Results Median age was 55.48 years (95% confidence interval (CI) 51.16–59.8), males 67.9%. History of cardiovascular diseases was in 54 (68.35%), diabetes mellitus in 18 (22.78%), hepatitis B and/or C in 31 (39.2%), intravenous drug dependency in 27 (34.18%), chronic kidney disease in 38 (48.1%), median Charlson comorbidity index was 5.44 (95% CI 4.52–6.37). Left-side IE was in 50 (63.29%), right-side IE – in 23 (29.12%), left-right-side IE in 6 (7.59%). Secondary IE was in 53 patients (62.3%). Embolic events were diagnosed in 27 cases (34.18%), in-hospital mortality – in 22 (27.8%). Microbiological method identified etiological agent in 55 of 85 cases (61.2%), featuring Staphylococcus aureus (n=23), Staphylococcus CoNS (n=6), Escherichia coli (n=1), Acinetobacter spp. (n=2), Streptococcus spp. (n=2), Enterococcus spp. (n=8), Klebsiella pneumoniae (n=2), Gemella haemolysans (n=2), several causative agents (n=6). Additional PCR testing identified etiology in 14 of 33 (42.2%) featuring Staphylococcus spp. (n=6), Enterococcus spp. (n=3), Streptococcus spp. (n=1), Aspergillus sp. (n=1) Pasteurella multocida (n=1), Enterococcus spp. + Staphylococcus spp. (n=1), Staphylococcus spp.+ A. baumanii + E. coli (n=1). PCR method identified 6 fals-positive results of microbilogical investigation [S. epidermidis (n=2), G. haemolysans, Acinetobacter spp., E. faecalis, K. pneumoniae], that are most probably due to preanalytical sample contamination Conclusions Introduction of PCR into the algorithm of IE etiological diagnostic increased validity of laboratory findings on 23.5%. True culture negative IE was present in 19 of 85 patients. Rate of mortality and complications in IE remains high. FUNDunding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Peoples' Friendship University of Russia (RUDN), Moscow, Russia

Synthesis and Antimicrobial Activities of 1-((5-Chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl)-N-ethylpyrrolidine-2-carboxamide

A new 1-((5-chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl)-N-ethylpyrrolidine-2-carboxamide was synthesized from methyl-1-[(5-chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl]pyrrolidine-2-carboxylate and ethylamine. The compound methyl-1-[(5-chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl]pyrrolidine-2-carboxylate was synthesized from methyl pyrrolidine-2-carboxylate and 5-chloro-4-chlorosulfonyl-1-ethyl-2-methyl-imidazole. The compounds were characterized based on FTIR, 1H, 13C NMR, and DEPT 135 analysis. Antimicrobial activities of the 1-((5-chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl)-N-ethylpyrrolidine-2-carboxamide against Gram-positive (methicillin-susceptible Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, and Bacillus subtilis), Gram-negative (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae), and Candida albicans were carried out using the standard microbiological method. The newly synthesized 1-((5-chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl)-N-ethylpyrrolidine-2-carboxamide had no activities against the tested organisms. Keywords: 1-((5-chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl)-N-ethylpyrrolidine-2-carboxamide; methyl-1-[(5-chloro-1-ethyl-2-methyl-1H-imidazol-4-yl)sulfonyl]pyrrolidine-2-carboxylate; L-proline; ethylamine.

Isolation Rate of Campylobacter Spp. and Detection of Virulence Genes of Campylobacter jejuni Across the Broiler Chain

The aim of the study was to identify the isolation rate of thermotolerant campylobacters in a small-scale broiler-meat production farm over a one-year period. The second deliverable of the study was to determine the potential virulence markers. The laboratory investigation was performed on 283 samples (cloacal swabs, caeca, carcass swabs) collected on three sampling points (farm, slaughter line, and cold storage). The isolates obtained with the conventional microbiological method were confirmed with multiplex PCR for identification of campylobacters. The presence of 10 virulence genes was analyzed in the C. jejuni isolates ( flaA, racR, virB11, dnaJ, wlaN, cadF, ciaB, cdtA, cdtB, cdtC). Out of 283 samples, 169 (59.7%) were confirmed as Campylobacter spp., 111 (39.2%) C. jejuni, and 43 (15.2%) C. coli. C. jejuni was the most prevalent in all sampling points. Campylobacter spp. showed a characteristically seasonal prevalence with the highest isolation rate during the warmer period of the year. We detected the cadF and ciaB genes in all C. jejuni isolates. The flaA gene was present in 50% of the examined strains. The cdt genes (cdtA, cdtB, and cdtC) were confirmed in 52.8%, 52.8%, and 47.2% of the C. jejuni strains, respectively. C. jejuni showed 15 profiles of virulence patterns with four predominant profiles.

COMPARISON AND EVALUATION OF PHARMACOPOEIAL METHODS FOR THE ASSESSMENT OF POTENCY OF ANTIBIOTICS

The detection and assessment of potency of antibiotics are crucial for the pharmaceutics. The valid methods for microbiological assays in pharmacopoeias are mainly based on statistical comparison of the data obtained by measuring the cidal activity resulting from the treatment of the antibiotic active ingredient in the composition of the pharmaceutics with the target microorganism. However, it was seen that there is no validated microbiological method for some active ingredients. Due to microbiological assays are indispensable methods for determining the potency of some active ingredient groups, the calculation of the potency is performed logarithmically. In either turbidimetric or chromatographic methods, the statistical evaluation of the sample is compared with the standard reference material. Analysis data obtained by chromatographic and chemical methods are linear peak areas and spectrophotometer readings. In microbiological methods, the data obtained from the analyzes performed to determine the potency of antibiotics are the inhibition zone diameters or turbidimetric turbidity data. In this study, above-mentioned microbiological assays are compared in the context of the main pharmacopoeias EP, USP, CP, IP and BP, and evaluated in terms of the chromatographic method and classical microbiological method. It has been observed that chromatographic and chemical methods are not available to determine the potency of some pharmaceutical products containing antibiotics. The examinations made reveal the difficulty of analyzing some active ingredient groups according to chemical and chromatographic methods. For this reason, the importance of method validation studies is increasing in order to analyze active substances that do not have alternative analysis methods with microbiological and chemical methods. In this study, all validated microbiological methods were investigated, and it was aimed to determine alternative methods to chromatographic and chemical methods. It was concluded that the realization of new microbiological methods to be validated by evaluating the methods in all differences would facilitate the study. Peer Review History: Received: 18 May 2021; Revised: 15 June; Accepted: 28 June, Available online: 15 July 2021 Academic Editor: Dr. Asia Selman Abdullah, Al-Razi university, Department of Pharmacy, Yemen, [email protected] UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.5/10 Average Peer review marks at publication stage: 7.5/10 Reviewer(s) detail: Dr. Iman Muhammad Higazy, National Research Center, Egypt, [email protected] Dr. George Zhu, Tehran University of Medical Sciences, Tehran, Iran, [email protected] Similar Articles: AN OVERVIEW ON PHARMACOPOEIAS IN THE WORLD AND MONOGRAPH ELABORATION TECHNIQUES

Ability of Yeast Metabolic Activity to Reduce Sugars and Stabilize Betalains in Red Beet Juice

To lower the risk of obesity, diabetes, and other related diseases, the WHO recommends that consumers reduce their consumption of sugars. Here, we propose a microbiological method to reduce the sugar content in red beet juice, while incurring only slight losses in the betalain content and maintaining the correct proportion of the other beet juice components. Several yeast strains with different metabolic activities were investigated for their ability to reduce the sugar content in red beet juice, which resulted in a decrease in the extract level corresponding to sugar content from 49.7% to 58.2%. This strategy was found to have the additional advantage of increasing the chemical and microbial stability of the red beet juice. Only slight losses of betalain pigments were noted, to final concentrations of 5.11 % w/v and 2.56 % w/v for the red and yellow fractions, respectively.

Viability and phenotypic heterogeneity of Rhodococcus Rhodochrous CNMN-AC-05 in the presence of fullerene C60

Introduction. In recent years, due to wide applications of nanotechnologies in various fields, the safety of nanomaterials has become a pressing issue. Fullerene C60 is not an exception. Research on the activity of microorganisms and their interaction with nanoparticles is of major importance, both for microorganisms and for the ecosystem as a whole. Material and methods. Fullerene C60 powder was purchased from Sigma-Aldrich. The object of study was R. rhodochrous CNMN-Ac-05 strain. The number of viable bacterial cells was estimated by colony-forming units (CFU). The morphological features of the rhodococci colonies have been described according to the usual microbiological method. Results. It was established that fullerene C60 in concentrations of 1-25 mg/L fullerene C60 stimulated the growth of R. rhodochrous by 2.4-2.8 times. As the concentration of fullerene C60 increased up to 50-100 mg/L, the multiplication and growth of rhodococci decreased by 29.5% and 38% respectively. In the presence of 1-10 mg/L fullerene C60 the rhodococci population remained homogeneous, being composed of 100% S type colonies. The increase of fullerene C60 concentration led both to the decrease in the CFU number and to the appearance of R type colonies, up to 1.3% of population. Conclusions. Fullerene C60 in concentrations 1-100 mg/L had no obvious toxic effect on the rhodococci strain. The optimum concentration is 10 mg/L. The concentrations higher than 25 mg/L led to the dissociation of rhodococcal population and diminution in the CFU counts, but not to the total inhibition.

Maternal Late-Pregnancy Serum Unmetabolized Folic Acid Concentrations Are Not Associated with Infant Allergic Disease: A Prospective Cohort Study

ABSTRACT Background The increase in childhood allergic disease in recent decades has coincided with increased folic acid intakes during pregnancy. Circulating unmetabolized folic acid (UMFA) has been proposed as a biomarker of excessive folic acid intake. Objective We aimed to determine if late-pregnancy serum UMFA and total folate concentrations were associated with allergic disease risk in the offspring at 1 y of age in a population at high risk of allergy. Methods The cohort consisted of 561 mother–infant pairs from Western Australia. To be eligible the infant had to have a first-degree relative (mother, father, or sibling) with a history of medically diagnosed allergic disease. Maternal venous blood was collected between 36 and 40 wk of gestation. Serum UMFA was measured by LC–tandem MS. Serum total folate was determined using a microbiological method with chloramphenicol-resistant Lactobacillus rhamnosus as the test organism, and was collected between 36 and 40 wk of gestation. UMFA concentrations were measured by tandem MS using stable isotope dilution; folate concentrations were determined using the microbiological method with standardized kits. Infant allergic disease outcomes of medically diagnosed eczema, steroid-treated eczema, atopic eczema, IgE-mediated food allergy, allergen sensitization, and medically diagnosed wheeze were assessed at 1 y of age. Results Median (IQR) concentrations for UMFA and serum folate were 1.6 (0.6–4.7) and 53.2 (32.6–74.5) nmol/L, respectively. Of the infants, 34.6% had medically diagnosed eczema, 26.4% allergen sensitization, and 14.9% had an IgE-mediated food allergy. In both adjusted and unadjusted models there was little evidence of association between UMFA or serum folate and any of the infant allergy outcomes. Conclusions In this cohort of children at high risk of allergic disease there was no association between maternal UMFA or serum folate concentrations measured in late pregnancy and allergic disease outcomes at 1 y of age.

Evolution of folate content during barley malt production

The aim of this research was to study the effect of micro- and industrial scale malting on the folate content of barley. Two malting barley varieties (one spring and one winter) were studied, applying the same technology. Furthermore, a roasting experiment was carried out at given temperatures for different time periods. The total folate content was determined by microbiological method. The folate content of the barleys was between 10.1 and 23.4 µg/100 g dry matter. For micro- and industrial scale, malting folate content increased 6.5–8-fold and 4–7-fold, respectively, during the malting process. An unexpected result was observed during industrial malting: the folate content increased during kilning by 18–35%, unlike micro scale malting, where a 15–20% decrease was observed. Results obtained during roasting showed that folic acid content did not decrease when roasted for 20 min at 100 °C, but it decreased linearly with increasing temperature. Folate is completely degraded in 20 min at 200 °C. It can be stated that barley malt can serve as a relatively good source of folate, but barley variety and malting technology have significant impact on it.