Analysis of a new phage, KZag1, infecting biofilm of Klebsiella pneumoniae: genome sequence and characterization.

BACKGROUND: This study investigates the effectiveness of the bacteriophage KZag1 against drug-resistant Klebsiella pneumoniae, aiming to assess its potential as a therapeutic agent. The novelty lies in the characterization of KZag1, a Myovirus with specific efficacy against multidrug-resistant K. pneumoniae strains. This highlights the significance of exploring alternative strategies, particularly phage therapy, in addressing biofilm-associated infections. METHODS: KZag1, characterized by a typical Myovirus structure with a 75 ± 5 nm diameter icosahedral head and a 15 ± 5 nm short tail, was evaluated in experimental trials against 15 strains of K. pneumoniae. The infection cycle duration was determined to be 50 min, resulting in an estimated burst size of approximately 83 plaque-forming units per colony-forming unit (PFU/CFU). Stability assessments were conducted within a pH range of 4 to 12 and temperatures ranging from 45°C to 60°C. Biofilm biomass reduction was observed, particularly at a multiplicity of infection (MOI) of 10. RESULTS: KZag1 demonstrated infection efficacy against 12 out of 15 tested K. pneumoniae strains. The phage exhibited stability across a broad pH range and at elevated temperatures. Notably, treatment with KZag1 significantly reduced K. pneumoniae biofilm biomass, emphasizing its potential in combating biofilm formation. Genomic analysis revealed a complete genome of 157,089 base pairs with a GC content of 46.38%, encompassing 203 open reading frames (ORFs) and a cysteine-specific tRNA sequence. Comparison with phage GP4 highlighted similarities, with KZag1 having a longer genome by approximately 4829 base pairs and a higher GC content by approximately 0.93%. Phylogenetic analysis classified KZag1 within the Myoviridae family. CONCLUSION: The efficacy of KZag1 against K. pneumoniae biofilm suggests its potential as a therapeutic candidate, especially for drug-resistant infections. Further clinical research is warranted to explore its synergy with other treatments, elucidate genomic traits, compare with Myoviridae phages, and understand its host interactions. These findings underscore the promising role of KZag1 in addressing drug-resistant bacterial infections.

Performance and nutrigenomics modulations in response to the inclusion of biologically treated date-palm mulch and enzyme mixture in the diets of growing rabbits.

This study was conducted to evaluate the effects of Allzyme addition on biologically-treated date-palm mulch (DPM) based diets for growing rabbits. DPM was treated by Trichoderma viride, Trichoderma reesi 230, Plorotus oysterous, and Phanaerochyte chrysosporium. Eighty rabbits were assigned to four groups: a control group, tDPM (10% tDPM inclusion of total diet), Allzyme (Allzyme supplementation), and tDPM + Allzyme (tDPM and Allzyme supplementation). The biological treatment resulted in a significant increase in crude protein and reductions in crude fiber. There was an interaction between tDPM and Allzyme at 9- and 10-week BW. The negative effects of tDPM on BW started at 8-week of age. The tDPM had unfavorable effects on slaughter and meat quality traits. The tDPM-by-Allzyme interaction affected total protein and globulin concentrations. However, blood glucose concentration was influenced by both tDPM and Allzyme. A significant tDPM effect was detected on the expression of INSR, GHSR, and IGF1 genes. However, the Allzyme effect was significant for PPARg and FASN genes. In conclusion, feeding tDPM negatively impacted rabbit's performance, however, Allzyme supplementation alleviated some of those effects. Accordingly, tDPM is recommended to be included in the diets of growing rabbits along with Allzyme supplementation.

Biosynthesis and Characterization of Silver Nanoparticles Produced by Phormidium ambiguum and Desertifilum tharense Cyanobacteria.

The world faces a challenge with the pervasion of multidrug-resistant bacteria that encourages scientists to develop and discover alternative, ecofriendly, and easy-to-produce new antibacterial agents. Our work is part of the greater effort of scientists around the world to achieve this goal by the biological synthesis of silver nanoparticles using cyanobacterial extracellular and intracellular components as nonchemical reducing agents. Two Egyptian cyanobacteria were isolated and identified according to 16S rRNA gene sequencing as Phormidium ambiguum and a novel species Desertifilum tharense. The sequences were deposited with accession numbers MW762709 and MW762710 for Desertifilum tharense and Phormidium ambiguum, respectively, in the GenBank. The results of UV-Vis analysis showed promising extracellular Ag-NPs synthesis by Desertifilum tharense and Phormidium ambiguum under light conditions. Therefore, these Ag-NPs were characterized and evaluated for antibacterial and antioxidant activity. TEM and SEM analyses revealed the spherical crystals with face-centered cubic structures and size range of 6.24-11.4 nm and 6.46-12.2 nm for Ag-NPs of Desertifilum tharense and Phormidium ambiguum, respectively. XRD and EDX results confirmed the successful synthesis of Ag-NPs in their oxide form or chloride form. The FTIR spectrum data confirmed the presence of hydroxyl and amide groups. Desertifilum tharense Ag-NPs displayed the largest inhibition zone that ranged from 9 mm against Micrococcus luteus ATCC 10240 to 25 mm against methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300. For Phormidium ambiguum Ag-NPs, the inhibition zone diameter was in the range of 9 mm to 18 mm. The biosynthesized Ag-NPs significantly inhibited the growth of medically important resistance-pathogenic Gram-positive and Gram-negative bacteria. The Ag-NPs of Phormidium ambiguum exhibited the highest scavenging activity of 48.7% when compared with that of Desertifilum tharense, which displayed 43.753%.

Bioethanol production from defatted biomass of Nannochloropsis oculata microalgae grown under mixotrophic conditions.

In order to improve the economic feasibility and environmental sustainability of microalgal bioethanol production, a nontoxic, copious agricultural waste, sugarcane bagasse aqueous extract (SBAE) was used for cultivating Nannochloropsis oculata microalga (NNO-1 UTEX Culture LB 2164) as potential sources of substitutes for traditional nutrition to reduce the costs in cultivation through acid digestion and enzymatic treatment before being fermented by Saccharomyces cerevisiae (NRRLY-2034). The primary target of this research was to find out the ethanol from hydrolysate of the defatted biomass of N. oculata grown mixotrophically on SBAE and CO2 as carbon sources. For acid hydrolysis (AH), the highest carbohydrate yield 252.84 mg/g DW has been obtained with 5.0% (v/v) H2SO4 at 121 °C for 15 min for defatted biomass cultivated mixotrophically on sugarcane bagasse aqueous extract (SBAE) regarding 207.41 mg/g DW for defatted biomass cultivated autotrophically (control treatment). Whereas, the highest levels of reducing sugars has been obtained with 4.0% (v/v) H2SO4 157.47±1.60 mg/g DW for defatted biomass cultivated mixotrophically compared with 135.30 mg/g DW for the defatted control treatment. The combination of acid hydrolysis 2.0% (v/v) H2SO4 followed by enzymatic treatment (AEH) increased the carbohydrate yields to 268.53 mg/g DW for defatted biomass cultivated mixotrophically on SBAE regarding 177.73 mg/g DW for the defatted control treatment. However, the highest levels of reducing sugars have been obtained with 3.0% (v/v) H2SO4 followed by enzyme treatment that gave 232.39±1.77 for defatted biomass cultivated mixotrophically on SBAE and 150.75 mg/g DW for the defatted control treatment. The sugar composition of the polysaccharides showed that glucose was the principal polysaccharide sugar (60.7-62.49%) of N. oculata defatted biomass. Fermentation of the hydrolysates by Saccharomyces cerevisiae for the acid pretreated defatted biomass samples gave ethanol yield of 0.86 g/L (0.062 g/g sugar consumed) for control and 1.17 g/L (0.069 g/g sugar consumed) for SBAE mixotrophic. Whereas, the maximum ethanol yield of 6.17±0.47 g/L (0.26±0.11 g/g sugar consumed) has been obtained with samples from defatted biomass grown mixotrophically (SBAE mixotrophic) pretreated with acid coupled enzyme hydrolysis.

Carbon nanotubes catalyzed UV-trigger production of hyaluronic acid from Streptococcus equi.

Hyaluronic acid (HA) has great importance in biomedical applications. In this work, a novel nanoparticle-based method that stimulates the hyaluronic acid (HA) production by the bacteria Streptococcus equi subsp. Zooepidemicus has been reported. CNTs with diameters of 40-50 nm and lengths of about 20 mm were used at four different concentrations (0, 10, 25, 50, and 100 µg) to the bacteria and determined the mass of the produced HA in dependence on the exposure time under UV-irradiation. The results clearly showed that the exposure for one minute with low power UV light (254 nm) and 100 µg (CNTs) treatments steadily increased HA production from the control (0.062 g/L) to the highest value (0.992) g/L of HA. The incubation of the streptococci with CNTs led to an increase of the HA production by a factor of 4.23 after 300S exposure time under UV light, whereas the HA production was no significant enhancement under visible light. It is explained that the CNTs nanoparticle-stimulated increase of the HA production with the internalization of the nanoparticles by the bacteria since they "serve as co-enzymes" under induced mutation by UV-irradiation. Transformation process was carried out and showed that the major protein band of Streptococcus equi was observed in the Streptococcus DH5α. RAPD analysis indicates that the amplified DNA fragments and the percentage of polymorphism was similar between Streptococcus equi and Streptococcus DH50α. The chemical structure and molecular weight of the photoproduced HA from Streptococcus equi was similar to the chemical structure of the standard sample.