Characterization and optimization of exopolysaccharide extracted from a newly isolated halotolerant cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1 with antiviral activity.

Several antiviral agents lost their efficacy due to their severe side effects and virus mutations. This study aimed to identify and optimize the conditions for exopolysaccharide (EPS) production from a newly isolated cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1, besides exploring its antiviral activity. The cyanobacterial EPS was purified through DEAE-52 cellulose column with a final yield of 83.75%. Different analysis instruments were applied for EPS identification, including Fourier-transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), and gas chromatographic-mass spectrometry (GC-MS). Plackett-Burman's design demonstrated that working volume (X1), EDTA (X2), inoculum size (X3), CaCl2 (X4), and NaCl (X5) are the most important variables influencing EPS production. Central composite design (CCD) exhibited maximum EPS yield (9.27 mg/mL) at a working volume of 300 mL in a 1 L volumetric flask, EDTA 0.002 g/L, inoculum size 7%, CaCl2 0.046 g/L, and NaCl 20 g/L were applied. EPS showed potent antiviral activities at different stages of herpes simplex virus type-1 and 2 (HSV-1, HSV-2), adenovirus (ADV) and coxsackievirus (A16) infections. The highest half-maximal inhibitory concentration (IC50) (6.477 µg/mL) was recorded during HSV-1 internalization mechanism, while the lowest IC50 (0.005669 µg/mL) was recorded during coxsackievirus neutralization mechanism.

Production and characterization of polyhydroxybutyrate bioplastic precursor from Parageobacillus toebii using low-cost substrates and its potential antiviral activity.

There is an increasing desire for bioplastics produced from renewable resources as an alternative to their petrochemical counterparts. These biopolymers have long-unnoticed antiviral properties. This study aimed to produce and characterize bioplastics by Parageobacillus toebii using low-cost substrates and determine their antiviral activity against coxsackievirus B4. Seven low-cost substrates (bagasse, water hyacinth, rice straw, rice water, sesame husks, molasses, and corn syrup) were compared with glucose for bioplastic precursor production. The highest bioplastic produced was from water hyacinth and glucose, followed by molasses, rice straw, rice water, sesame husks, and bagasse. Water hyacinth and glucose media were further optimized to increase the bioplastic precursor yield. The optimization of the media leads to increases in bioplastic precursor yields of 1.8-fold (3.456 g/L) and 1.496-fold (2.768 g/L), respectively. These bioplastics were further characterized by thermogravimetric analysis (TGA), Fourier-transformed infrared (FTIR) spectroscopy, proton nuclear magnetic resonance (1H NMR), and gas chromatography-mass spectrometry (GC-MS). They are thermostable, and their characterizations confirm the presence of polyhydroxybutyrate. The antiviral assay showed reasonable antiviral effects for bioplastics from water hyacinth (80.33 %) and glucose (55.47 %) media at 250 µg/mL maximum non-toxic concentrations (MNTC). The present investigation demonstrates a low-cost model for producing polyhydroxybutyrate bioplastic precursor for antiviral applications.

Identification and statistical optimization of a novel alginate polymer extracted from newly isolated Synechocystis algini MNE ON864447 with antibacterial activity.

Cyanobacteria are a potential source of promising secondary metabolites with different biological activities, including antibacterial, antiviral, antifungal, antiprotozoal, and anticancer activities. To combat the emergence of antibiotic resistance, there is an urgent requirement for new drugs, and cyanobacteria metabolites can constitute alternative new antibacterial medication. The chemical complexity of their exopolysaccharides indicates that they have the potential to be bioactive molecules with many biological activities. The present study aimed to produce and optimise a novel alginate polymer from a newly isolated cyanobacterium, S. algini MNE ON864447, in addition to its promising antibacterial activity. We successfully isolated a new cyanobacterium strain, S. algini MNE ON864447 from the Nile River, which produces alginate as an extracellular polymeric substance. The isolated cyanobacterial alginate was identified using a set of tests, including FTIR, TLC, HPLC, GC-MS, and 1H NMR. Plackett-Burman statistical design showed that working volume (X1), the incubation period (X2), and inoculum size (X3) are the most significant variables affecting the production of alginate. The highest alginate production (3.57 g/L) was obtained using 4% inoculum size in 400 mL medium/L conical flask after 20 days of the incubation period. The extracted alginate showed potent antibacterial activity against both Gram-negative and Gram-positive bacteria and Streptococcus mutants (NCTC10449) are the most sensitive tested pathogen for purified cyanobacterial alginate with inhibition zone diameters of 34 ± 0.1 mm at 10 mg/mL of purified alginate while Vibro cholera (NCTC 8021) the lowest sensitive one and showed inhibition zone diameters of 22.5 ± 0.05 mm at the same cyanobacterial alginate concentration. This antibacterial activity is a critical step in the development of antibacterial drugs and presents a new challenge to fight against multi-resistant bacteria.

Emergence of microbial infections in some hospitals of Cairo, Egypt: studying their corresponding antimicrobial resistance profiles.

BACKGROUND: Antimicrobial resistance is one of the ten major public health threats facing humanity, especially in developing countries. Identification of the pathogens responsible for different microbial infections and antimicrobial resistance patterns are important to help clinicians to choose the correct empirical drugs and provide optimal patient care. METHODS: During the period from November 2020 to January 2021, one hundred microbial isolates were collected randomly from different specimens from some hospitals in Cairo, Egypt. Sputum and chest specimens were from COVID-19 patients. Antimicrobial susceptibility testing was performed according to CLSI guidelines. RESULTS: Most microbial infections were more common in males and in elderly people over 45 years of age. They were caused by Gram-negative, Gram-positive bacteria, and yeast isolates that represented 69%, 15%, and 16%, respectively. Uropathogenic Escherichia coli (35%) were the most prevalent microbial isolates and showed high resistance rates towards penicillin, ampicillin, and cefixime, followed by Klebsiella spp. (13%) and Candida spp. (16%). Of all microbial isolates, Acinetobacter spp., Serratia spp., Hafnia alvei, and Klebsiella ozaenae were extremely multidrug-resistant (MDR) and have resisted all antibiotic classes used, except for glycylcycline, in varying degrees. Acinetobacter spp., Serratia spp., and Candida spp. were secondary microbial infections in COVID-19 patients, while H. alvei was a bloodstream infection isolate and K. ozaenae was recorded in most infections. Moreover, about half of Staphylococcus aureus strains were MRSA isolates and reported low rates of resistance to glycylcycline and linezolid. In comparison, Candida spp. showed high resistance rates between 77 and 100% to azole drugs and terbinafine, while no resistance rate towards nystatin was reported. Indeed, glycylcycline, linezolid, and nystatin were considered the drugs of choice for the treatment of MDR infections. CONCLUSION: The prevalence of antimicrobial resistance in some Egyptian hospitals was high among Gram-negative, Gram-positive bacteria, and candida spp. The high resistance pattern -especially in secondary microbial infections in COVID-19 patients- to most antibiotics used is a matter of great concern, portends an inevitable catastrophe, and requires continuous monitoring to avoid the evolution of new generations.

Nostoc muscorum is a novel source of microalgal lectins with potent antiviral activity against herpes simplex type-1.

In our survey for a new antiviral agent, two types of lectin were purified from Nostoc muscorum using both ion-exchange and affinity columns chromatography. Nostoc muscorum lectins (NMLs) are categorized based on their carbohydrate preference. Nostoc muscorum lectin-1(NML-1) exhibited a strict binding specificity for complex glycoproteins without linked carbohydrates, and the other displayed specificity for α- glycosides mannose polymers (NML-2) and was classified as a glycoprotein with 16.8% linked carbohydrates. NML-1 displayed a single band of 166 kDa on native-PAGE and two bands of 81 kDa and 85 kDa on SDS-PAGE, which confirmed the heterodimeric nature of this lectin. While NML-2 is a 50 kDa glycoprotein composed of 25 kDa subunits. Physical characterization of NML-1 displayed its stability at a higher temperature of 90 °C for 5 min and over a wide pH range (4-9), while MNL-2 displayed stability up to a temperature of 80 °C for 25 min and a pH range of 5-8. NML-1 didn't require metal ions for agglutination activity, while the activity of NML-2 was doubled by manganese ions. The antiviral activity of two lectins was assessed against herpes simplex type-1 (HSV-1) using a plaque assay which revealed that NML-1 inhibited HSV-1 infection at an early stage in contrast to NML-2 which exerted its antiviral effect at the late stage of infection. These results suggest that Nostoc muscorum is a unique lead for antiviral drug discovery as it is a novel source for antiviral lectins with different modes of action.

In vitro assessment of dual (antiviral and antitumor) activity of a novel lectin produced by the newly cyanobacterium isolate, Oscillatoria acuminate MHM-632 MK014210.1.

A novel lectin was purified from newly cyanobacterium isolate, Oscillatoria acuminate MHM-632 MK014210.1 using affinity chromatography with a molecular weight of 120 kDa under native-PAGE and 30 kDa on reducing-PAGE, represented tetramer nature of this lectin. Oscillatorial lectin showed stability at 60 °C for 30 min, pH-dependent, with the highest activities over the pH range of 6-8, and required zinc ions to express its full activity. Oscillatorial lectin is a glycan-binding protein with a neutral carbohydrate content of 7.0% as evaluated by the phenol-sulfuric acid method. Polyols and α- glycosides polymer of mannose sugar or sugars alcohol were completely inhibited oscillatorial lectin with MIC of 0.195 mM, while ß-glycosides sugars did not show any inhibition effect. The oscillatorial lectin has anti-proliferative activity against Huh-7 and MCF-7 cancer cells and inhibited their proliferation with EC50 values of 106.75 µg/ml and 254.14 µg/ml, respectively. Besides the anticancer effect, oscillatorial lectin also has potent antiviral activity against HSV-1 in a dose-dependent manner via virions neutralization and inhibition of viral replication with IC50 values of 90.95 ng/ml and 131.3 ng/ml, respectively. The unique carbohydrate affinity of oscillatorial lectin provides insight into its use as a promising candidate in many biotechnological applications, like fighting viral infection and combating cancer disease.Communicated by Ramaswamy H. Sarma.

The use of cyanobacterial metabolites as natural medical and biotechnological tools: review article.

Cyanobacteria are photosynthetic, Gram-negative bacteria that are considered one of the most morphologically diverse groups of prokaryotes with a chief role in the global nutrient cycle as they fixed gaseous carbon dioxide and nitrogen to organic materials. Cyanobacteria have significant adaptability to survive in harsh conditions due to they have different metabolic pathways with unique compounds, effective defensive mechanisms, and wide distribution in different habitats. Besides, they are successfully used to face different challenges in several fields, including industry, aquaculture, agriculture, food, dairy products, pollution control, bioenergy, and pharmaceutics. Analysis of 680 publications revealed that nearly 1630 cyanobacterial molecules belong to different families have a wide range of applications in several fields, including cosmetology, agriculture, pharmacology (immunosuppressant, anticancer, antibacterial, antiprotozoal, antifungal, anti-inflammatory, antimalarial, anticoagulant, anti-tuberculosis, antitumor, and antiviral activities) and food industry. In this review, we nearly mentioned 92 examples of cyanobacterial molecules that are considered the most relevant effects related to anti-inflammatory, antioxidant, antimicrobial, antiviral, and anticancer activities as well as their roles that can be used in various biotechnological fields. These cyanobacterial products might be promising candidates for fighting various diseases and can be used in managing viral and microbial infections.Communicated by Ramaswamy H. Sarma.

Assessment of the Antibacterial Potential of Biosynthesized Silver Nanoparticles Combined with Vancomycin Against Methicillin-Resistant Staphylococcus aureus-Induced Infection in Rats.

Methicillin-resistant Staphylococcus aureus (MRSA) is considered one of the most serious multidrug-resistant bacteria worldwide. MRSA resistance to methicillin antibiotics made vancomycin, the acceptable treatment option. Silver nanoparticles (Ag-NPs) are among the well-known antibacterial substances showing multimode antibacterial action. Therefore, Ag-NPs are appropriate applicants for use in combination with vancomycin in order to augment its antibacterial action. This study aimed to biosynthesize silver nanoparticles and to evaluate its antibacterial activity against MRSA alone and when combined with vancomycin both in vitro and in vivo. Agaricus bisporus is used to reduce the silver nitrate salts in solution to yield silver nanoparticles which was characterized by UV-visible spectrophotometric analysis that shows maximum absorption at 420 nm as a preliminary confirmation for nanoparticles synthesis, Energy-Dispersive Analysis of X-ray (EDX) which confirms the crystalline nature of silver nanoparticles and transmission electron microscopy (TEM) image shows the particles in spherical form with mean size 27.45 nm. The synthesized silver nanoparticles were tested for antibacterial activity against MRSA, and the synergetic effects of the combination of silver nanoparticles and vancomycin were evaluated. The results showed a strong synergistic antibacterial effect between Ag-NPs and vancomycin in vitro with fractional inhibitory concentration 0.37 and in vivo against MRSA strain. The result revealed that mycosynthesized silver nanoparticles (NPs) enhance the in vitro and in vivo antibacterial activity of vancomycin against MRSA. These results suggested that sliver nanoparticles have an effective antibacterial activity against MRSA count, histopathology, and liver enzymes as well as protective immune response specially when combined with vancomycin in the lungs of infected rats with MRSA.

Identification of Novel Bioactive Compound Derived from Rheum officinalis against Campylobacter jejuni NCTC11168.

Gastric diseases are increasing with the infection of Campylobacter jejuni. Late stages of infection lead to peptic ulcer and gastric carcinoma. C. jejuni infects people within different stages of their life, especially childhood, causing severe diarrhea; it infects around two-thirds of the world population. Due to bacterial resistance against standard antibiotic, a new strategy is needed to impede Campylobacter infections. Plants provide highly varied structures with antimicrobial use which are unlikely to be synthesized in laboratories. A special feature of higher plants is their ability to produce a great number of organic chemicals of high structural diversity, the so-called secondary metabolites. Twenty plants were screened to detect their antibacterial activities. Screening results showed that Rheum officinalis was the most efficient against C. jejuni. Fractionation pattern was obtained by column chromatography, while the purity test was done by thin-layer chromatography (TLC). The chemical composition of bioactive compound was characterized using GC-MS, nuclear magnetic resonance, and infrared analysis. Minimal inhibitory concentration (MIC) of the purified compound was 31.25 µg/ml. Cytotoxicity assay on Vero cells was evaluated to be 497 µg/ml. Furthermore, the purified bioactive compound activated human lymphocytes in vitro. The data presented here show that Rheum officinalis could potentially be used in modern applications aimed at the treatment or prevention of foodborne diseases.

Biochemical characterization and application of a novel lectin from the cyanobacterium Lyngabya confervoides MK012409 as an antiviral and anticancer agent.

In the present study, a novel lectin was purified from the newly isolated cyanobacterium, Lyngabya confervoides MK012409 and tested for its antiviral and anticancer activity. Out of 30 isolates, Mabroka-s isolate which identified as Lyngabya confervoides MK012409 showed the highest agglutination titer. Lyngabyal lectin showed the greatest haemagglutination activity with pigeon/rabbit erythrocytes with a minimum concentration of 2.4 µg/ml. Physical characterization of Lyngabyal lectin showed ability to keep the activity at a higher temperature up to 80 °C with stability over a wide pH range (4-8) as well as its stability toward chemical denaturants. Carbohydrate specificity test revealed that the sugar alcohols completely inhibited the lectin haemagglutination activity. The electrophoretic analysis revealed that the lyngabyal lectin is a 140 kDa composed of two 70 kDa subunits. Lyngabyal lectin was able to inhibit the proliferation of MCF-7 and Caco-2 cancer cell lines with IC50 values of 246 ± 0.17 and 376.4 ± 0.34 µg/ml, respectively. Lyngabyal lectin also showed virucidal activity against HSV-1 with EC50 of 167 ± 0.52 ng/ml and inhibited plaque formation in the HSV-1 infected Vero cells with EC50 of 84.94 ± 0.34 ng/ml. These findings emphasize the ability of the lyngabyal lectin to fight breast and colon cancer besides it represents a promising antiviral agent.

Clinical and Environmental Surveillance of Rotavirus Common Genotypes Showed High Prevalence of Common P Genotypes in Egypt.

The objective of this study was to compare the prevalence of human rotavirus group A common G and P genotypes in human Egyptian stool specimens and raw sewage samples to determine the most common genotypes for future vaccine development. From 1026 stool specimens of children with acute diarrhea and using nested RT-PCR, 250 samples (24.37%) were positive for human rotavirus group A. Using multiplex RT-PCR, rotavirus common P and G genotypes were detected as 89.20% and 46.40% of the positive clinical specimens respectively. This low percentage of common G genotypes frequency may affect the efficiency of the available live attenuated oral rotavirus vaccines [Rotarix® (human rotavirus G1P[8]) and RotaTeq® (reassortant bovine-human rotavirus G1-4P[5] and G6P[8])], however the percentage of clinical specimens which were negative for common G genotypes but positive for P[8] genotype was 12.00%. From 24 positive raw sewage samples for rotavirus group A VP6 collected from Zenin and El-Gabal El-Asfar wastewater treatment plants (WWTPs), 21 samples (87.50%) were typeable for common P genotypes while 13 samples (54.17%) were typeable for common G genotypes. Phylogenetic analysis of a VP8 partial gene of 45 P-typeable clinical isolates and 20 P-typeable raw sewage samples showed high similarity to reference strains and the majority of mutations were silent and showed lower to non-significant similarity with the two vaccine strains. This finding is useful for determining the most common antigens required for future vaccine development.