Embracing innovation in microbiology education: Integrating online interactive simulation and F-2-F practical inquiry when teaching MADLI-TOF

Diagnostic laboratories routinely use Matrix-Assisted Laser Desorption/Ionisation-Time Of Flight (MALDI-TOF) to identify bacteria present in patient samples (Patel, 2013). MALDI-TOF quickly and accurately identifies organisms, yet, it is uncommon for undergraduate students to use this equipment; being limited to theoretical application only. To address this, we combined F-2-F laboratory classes with a MALDI-TOF simulation to teach students how to process clinical samples from start to finish. On completing the F-2-F and online simulation, students should understand the microbiological steps required to process diverse clinical samples; understand which follow-up tests will confirm the identity of the organism; understand how to prepare a sample for MALDI-TOF; and correctly analyse and interpret MALDI-TOF data. Third-year Infectious diseases students (n=37, mean age 22.2±4.9, mean GPA 5.46±0.84) at the University of South Australia participated in this study. In weeks 1 to 6 of semester, students learnt how to process clinical samples in a microbiology laboratory. In the mid-semester break, students used the simulation to process a patient sample (e.g. blood, sputum, or CSF). Students had to choose the correct culture medium, incubation conditions and interpret Gram stain results. Once isolated, single colonies were picked, processed and analysed using MALDI-TOF. The output was used to confirm the identity of the bacterial species. Written and video instructions were provided to teach students on use of the simulation. The MALDI-TOF data generated were interpreted while referring to laboratory data. On completion, students submitted a written laboratory report for assessment. Student feedback was obtained through a 5-point Likert-style questionnaire. Thematic analysis was performed on the free text written feedback. Students “agreed” that the simulation reinforced the logic required to analyse a patient sample (taught in the F-2-F sessions). They “agreed” that using the simulation in conjunction with F-2-F practicals was beneficial to their learning and that the simulation enhanced their ability to combine laboratory data (e.g. biochemical testing) with MALDI-TOF data. Prior to the simulation student understanding of MALDI-TOF was 2.6±0.54 (out of 5), increasing significantly to 3.1±0.55 after completing the simulation (Students t test; p < 0.0004). The mean score for the written report was 16.2±3.1 SD (max of 20), with only 2 students failing to achieve a minimum passing score. The combination of F-2-F laboratory and online MALDI-TOF effectively integrated all steps required to identify bacterial species in a clinical sample. While some aspects of navigating the simulation were identified as challenging for students, there were noted improvements in student understanding. We will continue to improve the flow of the simulation as well as the representation of culture plates for the different bacterial species. REFERENCE Patel R. (2013). Matrix-assisted laser desorption ionization-time of flight mass spectrometry in clinical microbiology. Clinical infectious diseases: an official publication of the Infectious Diseases Society of America, 57(4), 564–572. https://doi.org/10.1093/cid/cit24

Seasonal Variation of Residual Aluminum Concentration in Drinking Water

Data for aluminum concentration in Baghdad raw and drinking water have been taken from Baghdad Mayoralty for the period of the year (2005-2006) and for six water treatment plants (Al-Karkh, East Tigris, Al-Wathba, Al-Karama, Al-Qadisia and Al-Dora). The available data were analyzed by using statistical programs like (spss, statistica, grapher and excel) .Relation with water quality parameters was obtained like relation with pH and temperature. From the study the pH value was found to be ranged between (7.45-7.85) in supplied water for minimum aluminum residual. From the contour plot results show that an equation has been concluded which connect concentration with pH and temperature. Seasonal variation was also studied; maximum concentration found to be in summer season, at Al-Karama and Al-Wathba water treatment plants concentrations were constant during the period of the study. Comparison between water treatment plants was also made, Al-Qadisia and Al-Karkh water treatment plants have the best control conditions, aluminum concentrations were found to be within the Iraqi drinking water quality standards (0.2 mg/l)

Preparation of C-reactive protein latex reagent of human serum

C-reactive protein (CRP) is an acute phase reactant the concentration of which increases significantly following tissue injury or inflammation. It is present in the albumin fraction of the serum of patients and is antigenic upon injection into rabbits. We report here the development of a local latex reagent coated with rabbit anti-CRP. For this purpose CRP was isolated from pooled sera of patients and injected into rabbits. CRP rabbit antiserum was purified by using ammonium sulfate precipitation. Followed by passing on ion exchange chromatography (Diethylaminoethyl 'DEAE' cellulose). Then, latex particles were coated with purified antisera. The prepared CRP latex reagent agglutination was observed upon the addition of patients' sera and compared with that of normal sera. In addition, it was evaluated against a standard CRP latex reagent. The prepared reagent gave positive results (100%) with patients' sera and the positive control of the standard kit. While, the negative control of the standard kit and 30% of the normal sera gave negative results

The Way Digitalization is Impacting International Financial Markets: Stock Price Synchronicity

This paper investigates whether and how the development level of a country\u27s digital economy affects stock price synchronicity. The results indicate that countries with high levels of digital economy development exhibit low stock price synchronicity. Additionally, by decomposing stock price synchronicity into systematic and firm‐specific stock return variations, we find that systematic (firm‐specific) variations of stock returns decrease (increase) with the level of a country\u27s digitalization. These findings shed light on the future trend of stock price synchronicity in financial markets around the world and support the information‐based interpretation of stock price synchronicity

Seasonal Variations of Air Pollutants Concentrations within Baghdad City

Air pollution is increased significantly nowadays due to variousindustrial activities and car combustion emissions. In this work, data have beencollected from Al-Jadrya Monitoring Station (JMS). The effects of seasonalvariation on the pollutants concentration were examined. Furthermore,relations of nitrogen oxides concentrations (NOx) were assessed during the firsthours of the working days. Three randomly days from each of January,February, July and August have been taken to represent winter and summerseasons, respectively. It has been found that concentrations of all examinedpollutants have not exceeded the acceptable limits. However, nitrogen oxides(NOx) seem to be effected by seasonal variation where its concentration hasincreased in June and August of the summer season. The concentrations of otherpollutants (SO2, CO, PM) have not been influenced by seasonal variations, andthey depend on the gaseous source emissions at different times of the year.Hourly monitoring for nitrogen oxides (NOx) concentration showed increasingin concentrations during the summer season, especially in the early hours of theworking days

A functional genomics catalogue of activated transcription factors during pathogenesis of pneumococcal disease

Background: Association analysis is an alternative to conventional family-based methods to detect the location of gene(s) or quantitative trait loci (QTL) and provides relatively high resolution in terms of defining the genome position of a gene or QTL. Seed protein and oil concentration are quantitative traits which are determined by the interaction among many genes with small to moderate genetic effects and their interaction with the environment. In this study, a genome-wide association study (GWAS) was performed to identify quantitative trait loci (QTL)controlling seed protein and oil concentration in 298 soybean germplasm accessions exhibiting a wide range of seed protein and oil content. Results: A total of 55,159 single nucleotide polymorphisms (SNPs) were genotyped using various methods including illumina Infinium and GoldenGate assays and 31,954 markers with minor allele frequency >0.10 were used to estimate linkage disequilibrium (LD) in heterochromatic and euchromatic regions. In euchromatic regions, the mean LD (r2) rapidly declined to 0.2 within 360 Kbp, whereas the mean LD declined to 0.2 at 9,600 Kbp in heterochromatic regions. The GWAS results identified 40 SNPs in 17 different genomic regions significantly associated with seed protein. Of these, the five SNPs with the highest associations and seven adjacent SNPs were located in the 27.6-30.0 Mbp region of Gm20. A major seed protein QTL has been previously mapped to the same location and potential candidate genes have recently been identified in this region. The GWAS results also detected 25 SNPs in 13 different genomic regions associated with seed oil. Of these markers, seven SNPs had a significant association with both protein and oil. Conclusions: This research indicated that GWAS not only identified most of the previously reported QTL controlling seed protein and oil, but also resulted in narrower genomic regions than the regions reported as containing these QTL. The narrower GWAS-defined genome regions will allow more precise marker-assisted allele selection and will expedite positional cloning of the causal gene(s)

The variable region of the pneumococcal pathogenicity island 1 is responsible for the unusually high virulence of a serotype 1 isolate

Streptococcus pneumoniae is the leading infectious cause of death in children in the world. However, the mechanisms that drive the progression from asymptomatic colonization to disease are poorly understood. Two virulence-associated genomic accessory regions (ARs) were deleted in a highly virulent serotype 1 clinical isolate (strain 4496) and examined for their contribution to pathogenesis. Deletion of a prophage encoding a platelet-binding protein (PblB) resulted in reduced adherence, biofilm formation, reduced initial infection within the lungs, and a reduction in the number of circulating platelets in infected mice. However, the region’s overall contribution to the survival of mice was not significant. In contrast, deletion of the variable region of pneumococcal pathogenicity island 1 (vPPI1) was also responsible for a reduction in adherence and biofilm formation but also reduced survival and invasion of the pleural cavity, blood, and lungs. While the 4496�PPI1 strain induced higher expression of the genes encoding interleukin-10 (IL-10) and CD11b in the lungs of challenged mice than the wild-type strain, very few other genes exhibited altered expression. Moreover, while the level of IL-10 protein was increased in the lungs of 4496�PPI1 mutant-infected mice compared to strain 4496-infected mice, the levels of gamma interferon (IFN-�), CXCL10, CCL2, and CCL4 were not different in the two groups. However, the 4496�PPI1 mutant was found to be more susceptible than the wild type to phagocytic killing by a macrophage-like cell line. Therefore, our data suggest that vPPI1 may be a major contributing factor to the heightened virulence of certain serotype 1 strains, possibly by influencing resistance to phagocytic killing

The pneumococcal alpha-glycerophosphate oxidase enhances nasopharyngeal colonization through binding to host glycoconjugates

Streptococcus pneumoniae (the pneumococcus) is a major human pathogen, causing a broad spectrum of diseases including otitis media, pneumonia, bacteraemia and meningitis. Here we examined the role of a potential pneumococcal meningitis vaccine antigen, alpha-glycerophosphate oxidase (SpGlpO), in nasopharyngeal colonization. We found that serotype 4 and serotype 6A strains deficient in SpGlpO have significantly reduced capacity to colonize the nasopharynx of mice, and were significantly defective in adherence to human nasopharyngeal carcinoma cells in vitro. We also demonstrate that intranasal immunization with recombinant SpGlpO significantly protects mice against subsequent nasal colonization by wild type serotype 4 and serotype 6A strains. Furthermore, we show that SpGlpO binds strongly to lacto/neolacto/ganglio host glycan structures containing the GlcNAcβ1-3Galβ disaccharide, suggesting that SpGlpO enhances colonization of the nasopharynx through its binding to host glycoconjugates. We propose that SpGlpO is a promising vaccine candidate against pneumococcal carriage, and warrants inclusion in a multi-component protein vaccine formulation that can provide robust, serotype-independent protection against all forms of pneumococcal disease

Morfološke značajke krvnih stanica u klinički zdravih odraslih ljama (Lama glama).

Morphological characteristics, including numeration, differentiation and features of blood cells were determined in ten male and ten female clinically normal adult llamas. Haematological parameters included: erythrocyte count, haemoglobin concentration, packed cell volume, erythrocyte indices, reticulocyte count, platelet count, total leukocyte count and differential leukocyte count. It appeared that llama haemogram was characterized by the presence of numerous but small erythrocytes, high total leukocyte count and a high number of mainly immature eosinophils. Unique cellular morphological characteristics commonly observed in May-Grunwald-Giemsa stained blood smears were: folded erythrocytes, Cabot’s rings, hypersegmented neutrophil nuclei, granular lymphocytes and immature eosinophils.U 10 muških i 10 ženskih klinički zdravih odraslih ljama određivane su morfološke značajke, broj, razlike i osebujnost krvnih stanica. Određivan je broj eritrocita, koncentracija hemoglobina, hematokrit, obilježja eritrocita, broj retikulocita, broj trombocita, ukupni broj leukocita i diferenciranost leukocita. Čini se da je za hemogram ljama karakterističan velik broj eritrocita male veličine, zatim velik ukupni broj leukocita i velik broj uglavnom nezrelih eozinofila. Jedinstvene morfološke značajke stanica, promatrane u krvnim razmascima obojenima po May-Grünwald-Giemsi, bile su naborani eritrociti, Cabotovi prsteni, hipersegmentirane jezgre neutrofila, granulirani limfociti i nezreli eozinofili

Correction: Alhamami et al. First Emergence of Resistance to Macrolides and Tetracycline Identified in Mannheimia haemolytica and Pasteurella multocida Isolates from Beef Feedlots in Australia. Microorganisms 2021, 9, 1322

The authors wish to make the following corrections to this paper [1]: There are metadata errors in Supplementary Figure S2 and Table 8, which state that isolate 17BRD-035 was isolated from a feedlot in NSW when in fact it came from Queensland. Figure S2 in Supplementary Materials was changed accordingly and was included with a separate document: https://www.mdpi.com/article/10.3390/microorganisms9061322/s1 In Table 8, the ST column for strain P. m 17BRD-035 was changed from NSW to QLD, the correct Table 8 is as follows: Table 8. Resistance profile, RAPD pattern and presence of antimicrobial resistance genes among isolates of Pasteurella multocida (P. m) (n = 28) and Mannheimia haemolytica (M. h) (n = 1) +, present, −, absent. Table (see supplement) The authors apologize for any inconvenience caused and state that the scientific conclusions are unaffected. The original article has been updated