Plating Media Research Articles

Sitagliptin Extends Lifespan of Caenorhabditis elegans by Inhibiting Insulin/Insulin-Like Signaling and Activating Dietary Restriction-Like Signaling Pathways

Introduction: The discovery of longevity molecules that delay aging and prolong lifespan has always been a dream of humanity. Sitagliptin phosphate (SIT), an oral dipeptidyl peptidase-4 (DPP-4) inhibitor, is an oral drug commonly used in the treatment of type 2 diabetes (T2D). In addition to being antidiabetic, previous studies have reported that SIT has shown potential to improve health. However, whether SIT plays a role in the amelioration of aging and the underlying molecular mechanism remain undetermined. Methods: Caenorhabditis elegans (C. elegans) was used as a model of aging. Lifespan assays were performed with adult-stage worms on nematode growth medium plates containing FUdR with or without the specific concentration of SIT. The period of fast body movement, body bending rates, and pharyngeal pumping rates were recorded to assess the healthspan of C. elegans. Gene expression was confirmed by GFP fluorescence signal of transgenic worms and qPCR. In addition, the intracellular reactive oxygen species levels were measured using a free radical sensor H2DCF-DA. Results: We found that SIT significantly extended lifespan and healthspan of C. elegans. Mechanistically, we found that several age-related pathways and genes were involved in SIT-induced lifespan extension. The transcription factors DAF-16/FOXO, SKN-1/NRF2, and HSF-1 played important roles in SIT-induced longevity. Moreover, our findings illustrated that SIT-induced survival benefits by inhibiting the insulin/insulin-like signaling pathway and activating the dietary restriction-related and mitochondrial function-related signaling pathways. Conclusion: Our work may provide a theoretical basis for the development of anti-T2D drugs as antiaging drugs, especially for the treatment of age-related disease in diabetic patients.

Antimicrobial and Antioxidant Activity Screening of Mushrooms Growing in the Leningrad Region

Introduction. The emergence of new strains of microorganisms that are multidrug resistant (MDR) in relation to the antimicrobial drugs used is one of the pressing problems of modern medicine. To prevent an increase in MDR-related deaths, the search for new antibiotics and their introduction into medical practice must be continuously ongoing. Infectious diseases are also accompanied by cell damage and the development of free radical oxidation processes, therefore the search for new antioxidants is also an important task. Considering the powerful biosynthetic potential of basidiomycetes, this group of fungi has every prospect of becoming a new source of biologically active substances in general, as well as antibiotics and antioxidants in particular. Cap mushrooms, represented mainly by basidiomycetes, number about 14,000 species and are an accessible source of raw materials for the search for promising antimicrobial compounds and antioxidants.Aim. Study of the antioxidant and antimicrobial activity of total extracts obtained from cap mushrooms against Escherichia coli, Staphylococcus aureus and Candida albicans and assessment of the suitability of cap mushrooms as a natural source of substances with antimicrobial and antioxidant activity.Materials and methods. The antifungal and antibacterial activity of the extracts was determined by the micromethod of two-fold serial dilutions in a liquid nutrient medium in 96-well plates in duplicate. The study of this type of biological activity was carried out against reference (type) strains Staphylococcus aureus ATCC 6538-P, Escherichia coli ATCC 25922, Candida albicans NCTC 885-653. To study antioxidant activity using DPPH, we used alcoholic extracts from the fruiting bodies of mushrooms obtained by maceration with 96 % ethanol at a ratio of raw materials to extractant of 1 to 8 for 24 hours, an aqueous solution of ascorbic acid (vitamin C) and an ethanol solution of Trolox.Result and discussion. In relation to S. aureus, a representative of gram-positive flora, the studied extracts of cap mushrooms showed low activity, on average about 2500 or 5000 μg/ml. In relation to E. coli, a representative of gram-negative flora, 8 % of the studied cap mushroom extracts showed an average activity of about 1250 μg/ml. The largest number of cap mushroom extracts – 19% of all studied species – showed activity against the yeast micromycete C. albicans. The highest activity against C. albicans was observed in extracts of the mushrooms Cantharellula umbonata with an MIC of 625 μg/ml, Cortinarius olivaceofuscus with an MIC of 625 μg/ml, and Hypomyces chrysospermus with an MIC of 312 μg/ml. During screening of antioxidant activity, the studied extracts were divided into three groups: with high (more than 50 % PPR), medium (from 15 to 50 % PPR) and low (less than 15 %) antioxidant activity. It was shown that the sum of phenolic compounds significantly correlates with the level of antioxidant activity in all three groups, but in the groups with medium and low antioxidant activity there are also other non-phenolic groups of compounds that make a significant contribution to the total antioxidant activity.Conclusion. Cap mushrooms are a promising source of biologically active substances with antifungal and antioxidant activity.

Antifungal activities of leaf extracts from the Capsicum genus

The emergence of microorganisms resistant to antimicrobials has driven the search for new substances aimed at the control and prevention of pathogens. Plant antimicrobial peptides have biotechnological applications in the development of drugs and agricultural inputs for this purpose. The present work aims to evaluate the antifungal activity of extracts containing peptides obtained from the leaves of Capsicum plants. The accessions UENF 1626, 1632 and 1381 and the cultivar Carioquinha were used. The leaves were macerated, and then extraction was performed using a 60% methanol and dichloromethane solution in a 1:1 ratio. The hydromethanolic phase was separated and used for testing after evaporation of methanol. All extracts have a similar electrophoretic profile on tricine gel stained with Coomassie Blue, showing 2 bands of approximately 6.5 and 3.5 kDa. Inhibition of fungal growth with 24 h of culture was performed in liquid culture medium in 96-well plates with reading at 620 nm using extracts with a protein concentration of 200 mg mL-1. The filamentous fungi Fusarium solani, Fusarium lateritium, and Colletotrichum lindemuthianum and the yeasts Candida albicans, Candida buinensis, Candida tropicalis, Saccharomyces cerevisiae, Pichia membranifaciens and Kluyveromyces marxianus were used. The extract with the greatest antifungal potential was the leaves of Capsicum annuum cv. Carioquinha, inhibiting the growth of F. solani by 29%, C. lindemuthianum by 15%, Candida buinensis by 22% and K. marxianus by 24%. Therefore, the extract has peptides with biotechnological potential.

Prevalence of antibiotic-resistant gram-negative bacteria among patients with urinary tract infection at a facility in Ado Ekiti, Ekiti-State, Nigeria

Urinary Tract Infections (UTIs) constitute a huge public health burden especially in developing countries. This study was carried out to determine the current incidence of antibiotic resistance among Gram-negative bacteria recovered from urine of patients with UTI. This study used a cohort study design wherein one hundred and twenty one (121) mid-stream urine samples were collected and cultured onto sterile Cysteine Lactose Electrolyte Deficient medium (CLED) and MacConkey agar plates. All bacteria recovered were tested for resistance to the selected antibiotics as follows: ceftazidime (30 μg), ceftriaxone (30 μg), gentamicin (1 0μg), ampicillin (10 μg), meropenem (10 μg), ertapenem (10 μg), tetracycline (30 μg), norfloxacin (10 μg) and pefloxacin (5 μg). Twenty (20) isolates were chosen for detection of extended spectrum beta-lactamase producers using the double disk synergy test (DDST). All the bacteria were tested for biofilm production and haemolysis. Among the one hundred and twenty-one (121) mid-stream urine samples collected from patients, forty-six (46) samples grew significant bacteria. The bacteria recovered were Klebsiella pneumonia17 (37.0%), Klebsiella oxytoca 2 (4.3%), Escherichia coli (32.6%) and Proteus mirabilis (26.0%). Overall, the bacteria showed the highest resistance to pefloxacin (97.8%) and least resistance to meropenem (34.7%). The bacteria also showed high multiple resistance. All the bacteria subjected to DDST were confirmed as extended-spectrum beta-lactamase ESBL producers. Forty (43.6%) bacteria were designated as strong biofilm producers while 17.4 % bacteria showed hemolysis. The findings obtained in this study have ramifications for clinical management of UTI and public health public health in general. There should be strict enforcement of antibiotic stewardship.

Effects of Different Types of Probiotics on Microbial Colonization in Saliva of Orthodontic Patients—In Vivo Study

Introduction Streptococcus mutans and Lactobacillus count is found to be higher in orthodontic patients and is one of the leading causes of white spot lesion formation. Probiotics are emerging as alternative methods of plaque control. Aims and Objectives To determine the effects of different types of probiotics on S. mutans and Lactobacillus levels in saliva of orthodontic patients. Materials and Methods A total of 140 subjects were divided into Control group (Group I) with 20 subjects and study group taking probiotics (Group II) with 120 subjects. Saliva samples were taken at baseline ( T0), after three weeks ( T1), and after six weeks ( T2). The samples were diluted and incubated on specific agar media plates. The colonies observed were expressed in colony forming units per milliliter (CFU/ml) of saliva. Group II was further subdivided into six subgroups based on probiotic intake with 20 subjects in each subgroup. Results Statistically significant difference was found in salivary microbial counts in all the probiotic groups ( p < 0.05). The highest reduction in count of S. mutans (653.00 ± 102.23 CFU/ml to 84.1 ± 30.48 CFU/ml) and Lactobacilli (87.35 ± 15.64 CFU/ml to 14.60 ± 8.28 CFU/ml) was seen in the lozenges group. Conclusion Probiotics decrease the number of salivary S. mutans and Lactobacilli and lozenges could be considered for orthodontic fixed appliance patients.

Exact solution for thermal vibration of multi-directional functionally graded porous plates submerged in fluid medium

An analytical method for analyzing the thermal vibration of multi-directional functionally graded porous rectangular plates in fluid media with novel porosity patterns is developed in this study. Mechanical properties of MFG porous plates change according to the length, width, and thickness directions for various materials and the porosity distribution which can be widely applied in many fields of engineering and defence technology. Especially, new porous rules that depend on spatial coordinates and grading indexes are proposed in the present work. Applying Hamilton’s principle and the refined higher-order shear deformation plate theory, the governing equation of motion of an MFG porous rectangular plate in a fluid medium (the fluid-plate system) is obtained. The fluid velocity potential is derived from the boundary conditions of the fluid-plate system and is used to compute the extra mass. The Galerkin-Vlasov solution is used to solve and give natural frequencies of MFG porous plates with various boundary conditions in a fluid medium. The validity and reliability of the suggested method are confirmed by comparing numerical results of the present work with those from available works in the literature. The effects of different parameters on the thermal vibration response of MFG porous rectangular plates are studied in detail. These findings demonstrate that the behavior of the structure within a liquid medium differs significantly from that within a vacuum medium. Thereby, they offer appropriate operational approaches for the structure when employed in various mediums.

Numerical investigation of ice plate fractures upon rigid ball impact

The fracture process and breaking capability of ice cover impacted by high-speed projectiles before underwater explosion is an urgent subject of scientific research in ice engineering applications. This paper presents the results of numerical simulations of the dynamic fracture behaviour induced by the impact of a small spherical projectile on an ice plate. The proposed 3D rate-dependent peridynamics model is validated by the dynamic ring Brazilian disc testing of ice specimens. Good agreements between the present model and the previous experiments are obtained in ice breaking processes and damage patterns of thin, medium and thick ice plates. Several cases are analysed and discussed in terms of various impact velocities, ice thicknesses, and boundary constraint conditions. The simulations identify detailed fracturing characteristics that are not observed in experiments, including the sequence and shape of crack propagation in a short time interval, the crater formation and the damage evolution along the thickness direction. The proposed peridynamics computational model provides new understanding of the potential of using penetrator–ice block impact tests to observe the evolution of complex brittle damage and reveal the failure mechanism.

The new 222-nm far ultraviolet-C lowers bacterial contamination to endoscopists during esophagogastroduodenoscopy.

This study aimed to clarify the disinfectant efficacy of the 222-nm far ultraviolet-C (UV-C) during esophagogastroduodenoscopy using bacterial cultures. The endoscopists performed esophagogastroduodenoscopy wearing a gown with a tryptic soy agar medium plate on their epigastric region and were divided into two groups: 222-nm far UV-C irradiation (UV group) and non-UV irradiation (non-UV group). As a control group, tryptic soy agar medium plates were placed about 110 cm above the floor. The incidence of bacterial contamination was determined by positive bacterial culture. The cultured bacteria were identified by 16S rRNA sequencing. Additionally, the actual UV exposure dose was measured using the UV-indicator card which changed colors upon exposure to 222 nm far UV-C. The bacterial culture positivity in the UV group (5.03%) was significantly lower than that in the non-UV group (25.76%), p < 0.0001. Most of the bacteria identified in the UV and non-UV groups were normal constituents of the oral flora, including Streptococcus salivarius and Staphylococci. Conversely, pathogenic microbes were found in the control group. The actual exposure doses of 222-nm far UV-C at the endoscopists' face, neck, and epigastric region were 2.09±0.29, 5.89±0.49, and 7.36±0.58 mJ/cm2, respectively. The 222-nm far UV-C irradiation reduced bacterial contamination for endoscopists. It can be used with conventional physical coverings to provide more effective infection control.

MHD dissipative flow past a permeable vertical plate in a porous medium in the presence of constant heat, mass flux, and diffusion‐thermo effect

AbstractAn analytical study of heat and mass transport in magnetohydrodynamic convective flow over an indefinitely long porous plate, which is vertically immersed in the porous medium, is investigated when heat sink, diffusion‐thermo, and thermal radiation are present. A magnetic field with uniform strength has been applied transversely directed into the fluid region. Analysis of diffusion‐thermo impact within the flow problem with constant heat and constant mass flux in the presence of thermal radiation and heat sink is the novelty of the current work. By using the Eckert number E as the perturbation parameter, the regular perturbation approach is used to solve nondimensional governing equations. On the flow and transport characteristics, the effects of Prandtl number, Grashof number, Eckert number, Dufour number, Schmidt number, radiation, magnetic field, and heat sink parameter have been investigated graphically and in tabular form, respectively. The innovative aspect of the current study is the inclusion of the Dufour effect, in addition to constant heat and mass flux at the plate. The Dufour effect can be utilized to improve separation processes, such as distillation, extraction, and chromatography. When the Dufour number and radiation parameter increase, the fluid velocity increases, and while the heat sink parameter increases, it decreases. As dissipation increases, the velocity of the fluid also increases. The concentration of the fluid is enhanced as mass diffusivity increases. When the heat absorption sink grows, the temperature of the fluid decreases.

Screening for Fungicide Efficacy in Controlling Blackleg Disease in Wasabi (Eutrema japonicum).

Blackleg disease is devastating for wasabi (Eutrema japonicum) production, occurring at any time and everywhere within the main production area of the Sichuan Province, China. There have been very few studies on the chemical control of this disease. In this study, we isolated and identified a local popular strain of the pathogen Plenodomus wasabiae. The isolated fungus strain caused typical disease spots on the leaves and rhizomes upon inoculation back to wasabi seedlings. The symptoms of blackleg disease developed very quickly, becaming visible on the second day after exposure to P. wasabiae and leading to death within one week. We then evaluated the efficacy of ten widely used fungicides to screen out effective fungicides. The efficacy of the tested fungicides was determined through mycelial growth inhibition on medium plates. As a result, tebuconazole and pyraclostrobin were able to inhibit the mycelial growth of P. wasabiae, and the most widely used dimethomorph in local production areas produced the lowest inhibition activity (13.8%). Nevertheless, the highest control efficacy of tebuconazole and pyraclostrobin on wasabi seedlings was only 47.48% and 39.03%, respectively. Generally, the control efficacy of spraying the fungicide before inoculation was better than that after inoculation. An increase in the application concentration of the two fungicides did not proportionately result in improved performance. We cloned the full-length sequence of sterol 14-demethylase (CYP51) and cytochrome B (CYTB) of which the mutations may contribute to the possible antifungalresistance. These two genes of the isolated fungus do not possess any reported mutations that lead to fungicide resistance. Previous studies indicate that there is a significant difference between fungicides in terms of the effectiveness of controlling blackleg disease; however, the control efficacy of fungicides is limited in blackleg control. Therefore, field management to prevent wound infection and unfavorable environmental conditions are more important than pesticide management.

First report of Aloe root and stem rot caused by Phytophthora palmivora in Yunnan Province, China.

Aloe genus plants are perennial evergreen herb belonging to Liliaceae family which is widely used in food, medicine, beauty, and health care (Kumar et al. 2019). In August 2021, symptoms of root and stem rot was observed in approximately 20% of Aloe vera plantings in Yuanjiang County, Yunnan Province, China (23° 64' 53" N, 101° 99' 84" E). The most typical symptoms were stem and root rot, browning and necrosis of vascular tissues, gradual greening, and reddish-browning of leaves from bottom to top, abscission, and eventual plant death (Fig. S1). Therefore, to isolate and identify the pathogen, the plants showing the above symptoms were collected. The plant tissues were cut from the edges of root and stem lesions, followed by disinfection with 75% ethanol for 1 min, rinsed three times with sterilized distilled water, and cut into 3 × 3 mm small squares after excision of marginal tissues. The tissues were transferred to the oomycetes selective medium (Liu et al. 2022) and incubated at 28 °C in the dark for 3~5 days, and suspected colonies were purified. The colonies were then inoculated onto potato dextrose agar (PDA), V8-juice agar (V8), and oatmeal agar (OA) medium plates for morphological characteristics. Finally, 18 isolates with the same colonial and morphological characteristics were obtained from 30 lesioned tissue and one of them was named as ARP1. On PDA, V8 and OA medium plates, the ARP1 colonies were white. On PDA plate, the mycelia were dense and the colonies were petal-like; on V8 plate, the mycelia were cashmere and the colonies were radial or star-like. Whereas, on OA plate, the mycelia were cotton-like and the colonies were fluffy and radial (Fig. S2 A~C). Mycelium did not have septum with high branching and swelling. Sporangia were abundant, semi-papillate, varying in shape from ovoid-ellipsoid to long-ellipsoid, 18-26 × 45-63 μm (average: 22 × 54 μm, n = 30), sporangia released numerous zoospores from the papillate after maturation. The chlamydospores were spherical, 20-35 μm in diameter (average: 27.5 μm, n = 30) (Fig. S2 D~F). These morphological features were like those of the pathogenic species of the oomycetes (Chen et al. 2022). For the molecular characterization, the genomic DNA of the isolate was extracted using the cetyl trimethyl ammonium bromide method, and the translation elongation factor 1α (tef-1α) (Stielow et al. 2015), β-tubulin (β-tub) (Kroon et al. 2004) and internal transcribed spacer (ITS) (White et al. 1990) of isolated strain ARP1 were amplified using primer pairs EF1-1018F/EF1-1620R, TUBUF2/TUBUR1 and ITS1/ITS4, respectively. The tef-1α, β-tub genes and ITS region of ARP1 were directly sequenced and their sequence information was deposited in GenBank under accession numbers OQ506129, OQ506127 and OQ449628. ARP1 was clustered on the same evolutionary branch with Phytophthora palmivora (Fig. S3). To confirm the pathogenicity of ARP1, the main root of A. vera was wounded to 1 cm long and 2 mm deep with a scalpel blade followed by inoculation with 50 ml suspension of ARP1 zoospores at a concentration of 1 × 106 spores / ml per potted plant, and an equal volume of water as control. All inoculated plants were placed in the greenhouse at 28°C, 12 h / 12 h light / dark. After 15 dpi, the inoculated plants showed typical symptoms of wilted and drooping leaves and stem and root rot, same as observed in the field condition (Fig. S4). After inoculation with ARP1, a strain with the same morphological and molecular characteristics as the original isolate was re-isolated, confirming Koch's postulates. To our knowledge, this is the first report of P. palmivora causing root and stem rot of A. vera in the study region. This disease could be a potential risk for aloe production and therefore appropriate management measures should be taken.

Visual screening of potassium solubilizing bacteria for increasing the solubilization of potassium minerals

Research on potassium solubilizing bacteria (KSB) has recently gained attention, most of it has been focused on the size of the halo zone; however, information regarding KSB growth duration and their potential to reduce the use of chemical fertilizers for sustainable agriculture and to be used as a biofertilizer are still lacking. The aim of this study was to offer a complete, long-term observation of KSB growth. We monitored the growth of two KSB strains that were isolated from the rhizosphere for 7 days on Modified Aleksandrov Medium (MAM) plates. We specifically designed the observation times. These methods were used to distinguish KSB’s capability to solubilize potassium minerals. The results indicated that KSB-12 isolate was capable of producing organic acid and thus solubilizing potassium faster than KSB-19 isolate at 1-4 days of incubation. Interestingly, the solubilizing zone of the KSB-12 isolate decrease after 5 days of incubation, while KSB-19 isolate continued to grow, maintain cell numbers, produce organic acids, and continuously release potassium. Different types of volatile and non-volatile organic acids have an effect on this phenomenon. In conclusion, we provided a well-designed time series study regarding the potassium solubilization efficiency of KSB, determining the fastest and longest strains of potential KSBs.

Fungal inhibition by lactic acid bacteria (LAB) is modulated in vitro by cocoa fermentation-related conditions: towards a biocontrol of fungi in processing cocoa

Fungal growth impair strongly the quality of processed cocoa, leading to low crops value for farmers and manufacturers. The antifungal activity of lactic acid bacteria (LAB) from fermenting cocoa was analyzed and the conditions of fungal inhibition were evaluated. Fungi were isolated from stored fermented and dried cocoa beans on sabouraud plate medium. The results showed that, LAB strongly inhibit fungi isolated from fermented cocoa beans notably Mucors and Aspergilli, with Lactobacillus plantarum inducing the stronger inhibition whereas Leuconostoc mesenteroides produced a weaker inhibition. Acids production assayed by HPLC was found to be not related to antifungal activity, since LAB strains producing strong antifungal activity were not necessarly the best acid producers. Maximum fungal inhibition occured at 35 °C in Lactobacillus plantarum and Leuconostoc mesenteroides but also at 30 °C in Lactobacllus casei. Likewise, a narrow acidic pH range (3.0-4.0) allowed full expression of fungal inhibtion in LAB analyzed that decreased progressively toward pH 7.0 and failed at alkalin pH. However, sucrose even at high content (8%) was found to have no effect on antifungal activity of LAB, but its bioconversion product glucose and fructose decreased this activity when concentrations were set at 6 et 2 %, respectively. All together, the results suggest that, LAB may effectively exert fungal inhibition in a narrow timing of cocoa fermentation, with pulp contained sugars as metabolic regulators of this inhibition. These results may contribute to a better management of LAB as starter culture for an efficient inhibition of fungal growth and prevention of cocoa contamination from mycotoxins.&#x0D; Key Words : Antifungal activity ; lactic acid bacteria ; cocoa fermentation ; inhibition variation; pH and temperature ; sugars content.

Evaluation of Siderophore Production by Different Promising Microbial Isolates

Indian alkaline soils are deficient in iron due to high pH, high calcium carbonate, highly permeable coarse textured soils, and soils low in organic carbon content. This study is carried out to increase the iron use efficiency of soil by using promising microbial isolates. Production of siderophore by promising microbial isolates is a greatest invention which help in increasing uptake of iron. A short-term laboratory experiment was carried “To evaluate the siderophore productions by different promising microbial isolates using chrome azurol S assay technique”. Ten microbial isolates were inoculated on plate as well as in broth medium. Plates were observed for orange halo formation around the bacterial growth. The size of orange halo around the colony of each microbial isolates was measured. The broth cultures inoculated with inoculum, incubated at 28 ± 20C for 24-72 hr with constant shaking at 120 rpm. After the incubation, the culture broths were centrifuged at 10,000 rpm for 10 min. The cell free supernatants were subjected to quantitative estimation of siderophores by CAS-shuttle, absorbance was measured at 630 nm against a reference consisting of 0.5 ml of uninoculated broth and 0.5 ml of CAS reagent. Obtained results indicated that in both plate and broth assays medium inoculated with strain Pseudomonas fluorescens show significantly higher orange halo formation around the growth and maximum siderophore content in aliquot. Inoculation of this plant growth promoting microbial isolates will be the great achievement in replacing chemical fertilizers in agriculture.

Isolation and Characterization of Plant Growth Promoting Rhizobacteria form Raphanus sativus (Radish)

Rhizobacteria are present in rhizosphere region of plant root system, which enhance the plant growth by various way like biological nitrogen fixation, siderophore production, phosphate solubilization and phytohormone production. The soil samples were collected from rhizosphere region of Raphanus sativus, after enrichment rhizobacteria was isolated by serial dilution method, diluted sample were spread on respective solid agar media plates. Isolated rhizobacteria was identify by biochemical and molecular characterization methods. The isolated PGPRs was Bacillus subtilis which was showed phosphate solubilization activity and they were enhanced 30% more green gram seed germination. The method of current research is screening, isolation and biochemical characterization of rhizobacteria form rhizosphere region of Raphanus sativus. Phosphate solubilizing bacteria, solubilized the unavailable phosphate and provide to plant. The main purpose of this research paper is to widen the understanding of the role of phosphate solubilizing bacteria in crop production as biofertilizers.

Microbiological Assessment of Indoor Air Quality in Selected Patient Wards at a Tertiary Hospital in Nigeria

Aims: Patients are at higher risk of nosocomial infections from pathogenic microbes contaminating the indoor air of hospitals. This study evaluated selected patient wards at University of Calabar Teaching Hospital (UCTH), for bacterial and fungal contaminants. The study also determined the microbial contamination level when compared to recommended sanitary standards.&#x0D; Methodology: To determine microbial load, petri-plate gravitational settling method was used. The set-ups were exposed for one hour (at morning and noon) for passive sampling onto growth media plates, in accordance with the 1/1/1 scheme as stipulated by the index of microbial air contamination standard. Petri-plates were subsequently covered with lids and placed in a cold box. Samples were then transported to the laboratory for further evaluation. Sampling was performed in duplicates within three months, and further analysis were conducted based on standard protocols. Mean counts of bacteria and fungi was recorded and expressed as colony forming units (CFU/m3).&#x0D; Results: Microbial counts in all wards surveyed ranged from ‘intermediate’ to ‘high’ when compared to the sanitary standards of the European Commission for non-industrial premises. However, the counts were within the WHO acceptable sanitary standard (&lt;1000 CFU/m3). Eleven bacterial genera and seven fungal genera were identified. Isolated bacteria were Staphylococcus aureus (18.3%), CoNS (6.7%), Bacillus sp (13.3%), Streptococcus spp (6.7%), Corynebacterium sp (1.7%), and Micrococcus sp (1.7%), Pseudomonas aeruginosa (15%), Escherichia coli (10%), Klebsiella pneumoniae (10%), Acinetobacter baumannii (8.3%), Salmonella sp (5%) and Enterobacter sp (3.3%). Conversely, isolated fungal contaminants were A. niger (25.9%), A. flavus (20.4%), Penicillium sp (16.7%), Candida sp (9.3%), Rhizopus sp (3.7%), Mucor sp (7.4%), Cladosporium sp (14.8%), and Fusarium sp (1.9%).&#x0D; Conclusion: Regular monitoring of indoor air quality in health-care settings is encouraged to effectively guide infection prevention and control strategies and to limit nosocomial infections occurrence.

A comparative study for optimization of MALDI-TOF MS identification of filamentous fungi.

To evaluate and compare the performance of three commercial culture media, two filamentous fungi libraries, and two different protein extraction procedures in MALDI-TOF MS fungal identification. A total of 21 quality control samples were cultured on Sabouraud dextrose agar (SDA), ID fungi plate medium (IDFP), and Sabouraud gentamicin chloramphenicol 2 agar (SGC2). For four consecutive days, fungal growths were inoculated on a MALDI target plate both by using a direct transfer technique (DT) and by using a formic acid-ethanol protein extraction procedure (EEP). The MALDI-TOF MS-generated spectra were identified by the MBT Bruker library and the MSI database. Selective culture media (IDFP and SGC2) significantly outperformed the non-selective SDA medium. IDFP was superior to the SGC2 medium for dermatophyte identification. The EEP only demonstrated a benefit over DT in the underperforming SDA medium. The MBT Bruker library outperformed the MSI database in Aspergillus identification while the MSI database outperformed the MBT library in dermatophyte identification. For non-Aspergillus fungi, the libraries performed comparably. The results of our study show the necessity of using selective culture media (IDFP and SGC2) for fungal identification with MALDI-TOF MS and demonstrate no significant benefit of the formic acid-ethanol protein extraction technique in these media. Given the relative strengths and weaknesses of the MBT library and the MSI database, it might currently be beneficial to consider these libraries as complementary and employ both databases to achieve optimal fungal identification.

CRIOTOLERANCIA DE EMBRIONES BOVINOS CULTIVADOS INDIVIDUALMENTE EN UN MEDIO SUPLEMENTADO CON L-CARNITINA

&lt;p&gt;&lt;strong&gt;Background:&lt;/strong&gt; L-carnitine is a lipid metabolism enhancer and a potent antioxidant that prevents oxidative damage and improves cryotolerance of bovine embryos. &lt;strong&gt;Objective:&lt;/strong&gt; To determine the effect of L-carnitine during oocyte maturation on developmental competence and cryotolerance of single bovine embryo cultured. &lt;strong&gt;Methodology:&lt;/strong&gt; Embryos were produced in vitro using abattoir-derived cumulus-oocyte complexes (COCs). In experiment 1, two individual maturation, fertilization and culture systems were used in 24-well plates with 20 µL drops of medium covered with mineral oil and 96-well plates with 30 µL drops of medium. In experiment 2, oocytes were randomly distributed into two groups and single matured in 96-well plates in medium supplemented or not with 0.6mg/mL L-carnitine. On day 7 post fertilization, blastocysts were vitrified on solid surface in Fiberplug. Non-vitrified blastocysts were used as control. Embryonic survival after devitrification was determined by blastocysts re-expansion and hatching rate at 24 and 48 hours of post-devitrification culture. Total cell number and apoptotic rate by TUNEL-DAPI staining were used as quality and cryotolerance indicator. In both cases, cleavage and blastocyst rates were evaluated at 48 hours and 7 days post fertilization, respectively. &lt;strong&gt;Results:&lt;/strong&gt; No significant differences were found for embryonic development between single culture systems. There was no effect of L-carnitine supplementation during maturation on embryo development, but embryo survival had increased (P &amp;lt; 0.05) at 24- and 48-hours post devitrification. &lt;strong&gt;Implications:&lt;/strong&gt; Treatment with L-carnitine had increased (P &amp;lt; 0.05) post-thaw re-expansion rates (86.8 ± 3.0 vs 70.0 ± 4.4) and it was similar to non-vitrified control (89.7 ± 2.6). Mean cell number and apoptotic cell index, were similar for all treatment groups. &lt;strong&gt;Conclusion:&lt;/strong&gt; L-carnitine supplementation during maturation, does not improve division rate and subsequent development of single cultured embryos, however increases cryotolerance post devitrification.&lt;/p&gt;

A Cost-Effective and Straightforward Approach for Conducting Short- and Long-Term Biomonitoring of Gold Mine Waters

Gold mining pollution has long-lasting effects on the environment, particularly through acid mine drainage (AMD) and heavy metal contamination. Monitoring and assessing the impact of this pollution is crucial, as well as evaluating the effectiveness of remediation efforts. In our study, conducted in the gold mining area of Zlatna (GMAZ), western Romania, we utilised on-site measurements of water temperature, pH, electrical conductivity, and dissolved oxygen, along with the quantification of culturable aerobic bacteria and microfungi using ready-to-use media plates. We also examined the taxonomic richness of water invertebrates (TRWI) and the environmental features of the sites. Our study found significant negative impacts on the water biota in mining areas, with microbial abundance proving to be a reliable indicator of AMD pollution. While water invertebrates can also serve as indicators of mining effects, their abundance alone may not always accurately reflect pollution levels at every site. This multiple-factor analysis highlights the influences of water type, geological characteristics, air temperature, and precipitation on the structure of the aquatic biota. We observed a natural attenuation of mining pollution in the GMAZ in the last seven years. This study demonstrates that the quantification of microbiota, along with TRWI and basic physicochemical parameters, can offer a cost-effective alternative to expensive monitoring methods for assessing mining pollution.

Treatment of clinical laboratory sewage using a decentralized treatment unit and risk reduction for its reuse in irrigation using hybrid disinfection

The disinfection efficacy of standalone chlorine, UV and their combined approach (hybrid) was investigated for the coliform removal in BioKube 1 and 2 treated effluents collected from different environmental settings of clinical and domestic wastes. Chlorine and UV disinfection were applied to BioKube treated wastewater with doses from 0 to 4 mg L−1 and 0–166 mJ cm−2 respectively. Combined disinfection strategies were designed to reduce the dose of chlorine and UV and to exploit the synergistic effect of them. The culturable coliforms were enumerated in treated wastewater sample (control), immediately after (reduction), and 24 h post disinfections (regrowth) using culture media plating and colilert-18 media. Both the BioKube systems (1 and 2) were effective in achieving the strict norms of physicochemical parameters, but not following the coliform counts of treated effluent for reuse in irrigation. A hybrid strategy of chlorine followed by UV was found to be the most effective among various standalone and combination approaches for the removal of coliforms (>4 log ER or <1000 CFU/100 mL) from both the treated effluents. However, coliform present in treated effluent of BioKube 1 were resisting (regrowth) against all kind of applied disinfectants except chlorine followed by UV dose at or more than 0.5 mg L−1 + 41 mJ cm−2. Limited reports are available on hybrid disinfection approaches with decentralized packaged sewage treatment units and this study would help to adopt as an effective tertiary treatment strategy for reuse of treated sewage for irrigation while ensuring public health safety.