Convolution Neural Networks for Point-of-Care Diagnostics of Bacterial Infections in Blood

ABSTRACTA droplet of blood, when evaporated on a surface, leaves dried residue—the fractal patterns formed on the dried residues can act as markers for infection present in the blood. Exploiting the unique patterns found in the residues of a naturally dried droplet of blood, we propose a Point-of-Care (POC) diagnostic tool for detecting broad-spectrum of bacterial infections (such as Enterobacter aerogenes, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, Salmonella Typhi) in blood. The diagnosis process we propose is straightforward and can be performed with the following steps: A droplet of blood (healthy or infected) of volume range 0.5 to 2 μl is allowed to dry on a clean glass surface and is imaged using a conventional optical microscope. A computer algorithm based on the framework of convolution neural network (CNN) is used to classify the captured images of dried blood droplets according to the bacterial infection. In total, our multiclass model reports an accuracy of 92% for detecting six bacterial species infections in the blood (with control being the uninfected or healthy blood). The high accuracy of detecting bacteria in the blood reported in this article is commensurate with the standard bacteriological tests. Thus, this article presents a proof-of-concept of a potential futuristic tool for a rapid and low-cost diagnosis of bacterial infection in the blood.

ISOLATION AND IDENTIFICATION OF MICROBIAL FAUNA IN UTI PATIENTS

Urinary tract infection caused by bacteria leads to inflammation. It is an important infection which can cause a highly morbidity rates in both males and females. The aim of the present study was determine the percentage prevalence of the UTI among the general population and the percentage of the bacterial isolates in the Urine samples. Among 150 samples bacterial isolates were obtained which included Escherichia coli, Klebsiella pneumonia, Proteus vulgaris, Pseudomonas aeruginosa. The highest prevalence of specific bacteria was found to be Escherichia coli recorded among all the UTI patients to be (55.55%) the least percentage among the bacteria were Klebsiella pneumonia, Proteus vulgaris, Pseudomonas aeruginosa and Enterobacter aerogenes which recorded 16.66%, 5.55%, 11.11% and 11.11% respectively. Results of the study showed that of femaleshad UTI more than the males.

Correction: Enterobacter aerogenes ZDY01 inhibits choline-induced atherosclerosis through CDCA-FXR-FGF15 axis

Correction for ‘Enterobacter aerogenes ZDY01 inhibits choline-induced atherosclerosis through CDCA-FXR-FGF15 axis’ by Jinghui Tang et al., Food Funct., 2021, 12, 9932–9946, DOI: 10.1039/D1FO02021H.

Phytochemistry and In vitro Antimicrobial Activity of Five Plant Species against Nine Common Human Pathogens

Momordica charantia, Senna podocarpa, Senna alata, Ocimum gratissimum, and Sida acuta which have reportedly been used in folklore for the treatment of various diseases were studied for their antimicrobial activity. Crude methanolic extracts of the leaves were screened for phytochemicals after which they were tested in vitro for activity against clinical isolates of Staphylococcus aureus, Klebsiella oxytoca, Pseudomonas aeruginosa, Proteus vulgaris, Bacillus subtilis, Enterobacter aerogenes, Aspergillus niger, A, flavus and Candida albicans. Phytochemicals present in the extracts included saponins, alkaloids, anthraquinones, tannins, flavonoids and cardiac glycosides. These extracts also showed varying degrees of activity against tested organisms at a concentration range of 15.6 to 500 mg/ml. However, S. acuta showed activity against P. vulgaris, E. aerogenes and As. flavus only. The MIC of the extracts against test organisms ranged from 31.25 to 250 mg/ml. The significant antimicrobial susceptibility of the plant extracts against gram positive bacterial pathogens and some pathogenic yeasts, may not be due to the presence and synergistic interactions of secondary metabolites found in these plant extracts. This would have implications in health, particularly in developing countries where a singnificant percentage of the population are still using traditional plant extracts for health care. Dhaka Univ. J. Pharm. Sci. 20(2): 139-148, 2021 (December)

Detection of Quorum Sensing Signal Molecules, Particularly N-Acyl Homoserine Lactones, 2-Alky-4-Quinolones, and Diketopiperazines, in Gram-Negative Bacteria Isolated From Insect Vector of Leishmaniasis

Gram-negative bacteria are known to use a quorum sensing system to facilitate and stimulate cell to cell communication, mediated via regulation of specific genes. This system is further involved in the modulation of cell density and metabolic and physiological processes that putatively either affect the survival of insect vectors or the establishment of pathogens transmitted by them. The process of quorum sensing generally involves N-acyl homoserine lactones and 2-alkyl-4-quinolones signaling molecules. The present study aimed to detect and identify quorum sensing signaling molecules of AHLs and AHQs type that are secreted by intestinal bacteria, and link their production to their extracellular milieu and intracellular content. Isolates for assessment were obtained from the intestinal tract of Pintomyia evansi (Leishmania insect vector). AHLs and AHQs molecules were detected using chromatography (TLC) assays, with the aid of specific and sensitive biosensors. For identity confirmation, ultra-high-performance liquid chromatography coupled with mass spectrometry was used. TLC assays detected quorum sensing molecules (QSM) in the supernatant of the bacterial isolates and intracellular content. Interestingly, Pseudomonas otitidis, Enterobacter aerogenes, Enterobacter cloacae, and Pantoea ananatis isolates showed a migration pattern similar to the synthetic molecule 3-oxo-C6-HSL (OHHL), which was used as a control. Enterobacter cancerogenus secreted C6-HSL, a related molecules to N-hexanoyl homoserine lactone (HHL), while Acinetobacter gyllenbergii exhibited a migration pattern similar to 2-heptyl-4-quinolone (HHQ) molecules. In comparison to this, 3-oxo-C12-HSL (OdDHL) type molecules were produced by Lysobacter soli, Pseudomonas putida, A. gyllenbergii, Acinetobacter calcoaceticus, and Pseudomonas aeruginosa, while Enterobacter cloacae produced molecules similar to 2-heptyl-3-hydroxy-4-quinolone (PQS). For Pseudomonas putida, Enterobacter aerogenes, P. ananatis, and Pseudomonas otitidis extracts, peak chromatograms with distinct retention times and areas, consistent with the molecules described in case of TLC, were obtained using HPLC. Importantly, P. ananatis produced a greater variety of high QSM concentration, and thus served as a reference for confirmation and identification by UHPLC-MRM-MS/MS. The molecules that were identified included N-hexanoyl-L-homoserine lactone [HHL, C10H18NO3, (M + H)], N-(3-oxohexanoyl)-L-homoserine lactone [OHHL, C10H16NO4, (M + H)], N-(3-oxododecanoyl)-L-homoserine lactone [OdDHL, C16H28NO4, (M + H)], and 2-heptyl-3-hydroxy-4(1H)-quinolone [PQS, C16H22NO2, (M + H)]. Besides this, the detection of diketopiperazines, namely L-Pro-L-Tyr and ΔAla-L-Val cyclopeptides was reported for P. ananatis. These molecules might be potentially associated with the regulation of QSM system, and might represent another small molecule-mediated bacterial sensing system. This study presents the first report regarding the detection and identification of QSM and diketopiperazines in the gut sand fly bacteria. The possible effect of QSM on the establishment of Leishmania must be explored to determine its role in the modulation of intestinal microbiome and the life cycle of Pi. evansi.

A Prospective Study on Neonatal Sepsis in a Tertiary Hospital, Nepal

About 20 % of neonates develop sepsis and among them approximately 1% die due to sepsis-related causes. Bacterial pathogens are the commonest cause of neonatal sepsis which is either early-onset (<72 hours of age) or late-onset (>72 hours). Little is known about the epidemiology and antimicrobial susceptibility pattern of sepsis causing bacterial pathogens in Nepal. A prospective study was carried out among neonates suspected to have sepsis and admitted to Tribhuwan University Teaching Hospital from January to December 2016. Clinical suspicion of sepsis was made based on clinical findings and laboratory parameters, later confirmed by isolation of organisms in blood culture. Drug resistance pattern of Gram-positive and Gram-negative bacteria were studied by standard methods. Meropenem resistant Gram-negative bacteria were processed for the detection of β-lactamases and resistant genes were detected by X-pert Carba-R (Cepheid) Assays. Of 372 neonates with clinically suspected sepsis, 132 (35.4%) had blood culture positivity, with 47% early-onset and 53% late-onset sepsis. Coagulase-negative Staphylococcus aureus (CONS) was the most common (37.9%) etiological agent followed by Klebsiella pneumoniae (12.9%). Of all 132 isolates, 81 (61.3%) were Gram-positive of which 22 (27.2%) were multi-drug resistant (MDR), three (3.7%) were methicillin-resistant S. aureus (MRSA), and 14 (17.2%) were methicillin-resistant CoNS; and 50 (37.8%) were Gram-negative of which 26 (52%) were MDR and 29 (58%) were resistant to β-lactamases. The blaKPC gene was detected in four isolates of K. pneumoniae, two of E. coli, one ABC (Acinetobacter baumanii complex), and one Enterobacter aerogenes whereas blaNDM gene was detected in one isolate of K. pneumoniae, two of E. coli, two Pseudomonas aeruginosa, one Acinetobacter baumanii complex, and one Enterobacter aerogenes. Overall mortality due to sepsis-related causes was 7.6% (10 of 132). One-third of clinically suspected neonatal sepsis cases were culture positive. Late-onset sepsis was more common than early onset. CoNS was the predominant bacterial isolate followed by Klebsiella pneumoniae, with high rates of multi-drug resistance.

Growth Optimization of Glyphosate-based Herbicides Utilizing Bacteria isolated from Lotic Water of Ogini Stream, Nigeria

Glyphosate-based herbicides are often used for the control of weeds grown on agricultural fields or farms. Different health problems have been reported to be associated with the use of glyphosate-based herbicides mainly due to their toxicity level. Thus, finding glyphosate utilizing microorganisms to remediate the glyphosate-based herbicides in the environment is crucial. The culture conditions for maximum utilization of glyphosate by bacterial isolates, Stenotrophomonas maltophilia, Bacillus cereus and Enterobacter aerogenes previously isolated from Ugini stream close to corn fields treated with glyphosate-based herbicide at Ofagbe, Delta State, Nigeria were optimized using mineral salt medium containing glyphosate as carbon source. The varied culture parameters assessed were temperature (30, 37 and 40 oC), pH (5, 6, 7, 8 and 9), initial glyphosate concentration (1, 3, 5, 7 and 9 g/L) and incubation time (2-14 days). Optical density (OD) at 560 nm of the culture was used to estimate cell growth or cell load of the glyphosate utilizing bacteria strains at every 2 days for 14 days. The following optimal conditions were determined: initial pH 9.0, incubation temperature 30 °C, initial concentration of glyphosate (1g/L) and incubation time of 12 days. Of the isolates on the medium containing the herbicide as sole carbon and energy source, Bacillus cereus showed the highest growth level (OD average, 0.127, pH average, 6.26. This was followed by Stenotrophomonas maltophilia (OD average = 0.114, pH average = 6.44) and Enterobacter aerogenes (OD average = 0.100, pH average, 6.56). At the increased of glyphosate in the medium there was decreased in growth of the bacteria. Bacillus cereus, Stenotrophomonas maltophilia and Enterobacter aerogenes indicated a high capacity to be able to degrade glyphosate. It is therefore concluded that the bacteria employed in this research can be recommended for bioremediation of environments contaminated with this chemical and further research should conducted to ascertain the catabolic genes present in these individual glyphosate degrading bacteria.