An assessment of efficacy of Iodine complex (Renessans) against SARS-CoV-2 in non-human primates (Rhesus macaque)

Renessans is an iodine complex which has proven in vitro antiviral activity including Anti-SARS-CoV-2 activity. The present study was designed to determine its efficacy against SARS-CoV-2 in monkeys (Rhesus macaque). A total of 14 monkeys were divided into four groups: A) Prophylactic group (n=03), (B) Treatment group (n=03), (C) infection control group (n=04) and (D) negative control group (n=04) and were housed in BSL-3 Animal facility while group D was housed at another animal house. Group A was administered with Renessans @ 2.85 mg/7 kg from 5 days prior to the infection to 08 days post infections (DPI). Group B was administered with Renessans from 03-08 DPI @ 2.85 mg/7 kg. Group C was administered with WIF only. The infection @ 2 x 106 TCID of SARS-CoV-2 was given to all group monkeys through intranasal and oral route under anesthesia. Nasal swab samples (at different times) and fecal matter on daily basis were collected for the detection of SARS-CoV-2 through real-time quantitative PCR. Three monkeys (one from each of group A, B and C) were euthanized at 07 DPI to determine the gross pathological lesions and SARS-CoV-2 detection from internal tissues. Nasal swabs from all the monkeys from group A, B and C were positive for SARS-CoV-2 at 02 and 07 DPI (Day 05 of treatment). At 14 DPI, all (100%) nasal swabs from group A were negative for SARS-CoV-2 while 50% and 100% were positive from group B and C, respectively. At 21 DPI, monkeys from group B were negative and all in group C were still positive for SARS-CoV-2. Similarly, fecal matter of monkeys in group A and B was returned negative in significantly lesser time as compared to monkeys from infection control group. Based on these research findings it is concluded that the Renessans has in-vivo SARS-CoV-2 activity and may result in early clearance of SARS-CoV-2. Therefore, a clinical trial of the drug in COVID-19 patients may reveal its anti-COVID-19 potential.

A longitudinal survey for genome-based identification of SARS-CoV-2 in sewage water in selected lockdown areas of Lahore city, Pakistan; a potential approach for future smart lockdown strategy

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections has affected more than 15 million people and, as of 22 July 2019, caused deaths of more than 0.6 million individuals globally. With the excretion of SARS-CoV-2 in the stool of symptomatic and asymptomatic COVID-19 patients, its genome detection in the sewage water can be used as a powerful epidemiological tool to predict the number of positive cases in a population. This study was conducted to detect SARS-CoV-2 genome in sewage water during the lockdown. Sewage samples, from 28 pre-selected sites, were collected on alternate days from 13-25 July, 2020 from two selected areas [Johar Town (n = 05) and Township (n = 23)], where smart lockdown were implemented by the government authorities on 9th July, 2020. Genomic RNA was extracted and the SARS-CoV-2 was detected and quantified using commercially available kit through Real-Time PCR. Out of 28, sixteen samples were positive on day one while 19, 17, 23, 17, 05 and 09 samples were positive on day 3, 5, 7, 9, 11, and 13, respectively. Results revealed a decreased positivity rate and SARS CoV-2 genome copies in sewage towards the end of lockdown however few sampling sites did not follow a clear pattern indicating the complexities in sewage water based surveillance i.e time of sampling etc. Hourly sampling from two sites for 24 hours also revealed the impact of sampling time on detection of SARS-CoV-2 genome in sewage. Results of current study insinuate a possible role of sewage-based COVID-19 surveillance in monitoring and execution of smart lockdowns.

An in vitro assessment of anti-SARS-CoV-2 activity of oral preparations of iodine complexes (RENESSANS)

Since the emergence of CoVID-19 pandemic in China in late 2019, scientists are striving hard to explore non-toxic, viable anti-SARS-CoV-2 compounds or medicines. We determined In Vitro anti-SARS-CoV-2 activity of oral formulations (syrup and capsule) of an Iodine-complex (Renessans). A monolayer of vero cells were exposed to SARS-CoV-2 in the presence and absence of different concentrations (equivalent to 50, 05 and 0.5 g/ml of I2) of Renessans. Anti-SARS-CoV-2 activity of each of the formulation was assessed in the form of cell survival, SARS-CoV-2-specific cytopathic effect (CPE) and genome quantization. With varying concentrations of syrup and capsule, a varying rate of inhibition of CPE, cells survival and virus replication was observed. Compared to 0.5 g/ml concentration of Renessans syrup, 5 and 50 g/ml showed comparable results where there was a 100% cell survival, no CPEs and a negligible viral replication ({Delta}CT= 0.11 and 0.13, respectively). This study indicates that Renessans, containing iodine, may have potential activity against SARS-CoV-2 which needs to be further investigated in human clinical trials.

Outer membrane proteins and lipopolysaccharides mediated antibody response against bovine Pasteurella multocida type B-2

Aims: The activation of cellular and humoral immunity depends upon nature of antigens. Complex proteins like bacterial outer membrane proteins (OMP) usually successfully activate both humoral and cellular immunity. Whereas antigens like bacterial lipopolysaccharides (LPS) usually elicit T-independent immunity i.e. humoral immunity without the activation of cellular immune wing. The present study was under taken to evaluate the comparative immunologic behavior of both the important molecules (bacterial lipopolysaccharide and outer membrane proteins) of Pasteurella multocida alone and in combination in bovine calves in field conditions. Methodology and results: Pasteurella multocida was isolated, purified and identified from an outbreak by mean of culture and biochemical methods. The pathogenicity of the confirmed isolates was done in rabbits (Oryctolagus cuniculus) on the principles of Koch’s postulates. Alum based vaccine against P. multocida was prepared and antibody titer against Outer membrane protein (OMP) and lipopolysaccharides (LPS) were determined by complement fixation test (CFT). The results showed that the antibody titer against OMP and LPS in whole culture vaccine is significantly higher than the respective tested vaccines. These results concluded that OMP no doubt is an active T-dependent immunogenic molecule but its immunogenicity increases many times when combined with LPS in whole culture vaccine. Conclusion, significance and impact of study: Lipopolysaccharides (LPS) in combination with outer membrane proteins (OMP) synergistically boost the humoral immune response in vaccinated animal.

Optimization of PCR for rapid detection of CTX-M gene in ESBL producing Klebsiella pneumoniae clinical isolates from Punjab, Pakistan

Aims: The study was carried out firstly, to evaluate the prevalence of extended spectrum beta lactamase (ESBL), multidrug resistant Klebsiella pneumoniae isolates from Punjab, Pakistan and secondly, to characterize the genotypes of their beta lactamase producing enzymes and optimization of PCR based method for rapid and authentic detection of antibiotic resistant gene. Methodology and results: Two hundred of K. pneumonia strains were isolated from different clinical samples. Blood and MacConkey Agar were used to isolate and identify bacterial microorganisms while Muller Hinton Agar was used to evaluate the antimicrobial susceptibility against different antibiotics as per CLSI 2012 guidelines. ESBL producing bacteria were screened by double disk synergy /combination disk test. PCR was optimized and performed for resistant gene (CTX-M). The results showed that most of the isolates were resistant to multiple antibiotic including cephalosporin, aztreonam, sulphamethoxazole/trimethoprim, ciprofloxacin, doxycyclin and were sensitive to imipenam and amikacin. Frequency of CTX-M gene in ESBL producing K. pneumoniae was 94%. Conclusion, significance and impact of study: Based on the finding of this study it is suggested that prevalence of CTX-M gene (95%) is very high among ESBL producing isolates. Therefore, PCR based method may help clinicians for rapid detection and treatment of patients by choosing right medication against the resistant bacteria as early as possible.

Evaluation of antifungal and antioxidant potential of two medicinal plants:Aconitum heterophyllum and Polygonum bistorta

Objective:To focus on the evaluation of antimicrobial and antioxidant activity of two endangered medicinal plants Aconitum heterophyllum (A. heterophyllum) and Polygonum bistorta (P. bistorta). Materials: Plant extracts were obtained by using microwave assisted extraction method. The in vitro antifungal activity of A. heterophyllum and P. bistorta extracts were determined by measuring diameters of inhibitory zones of these extracts against Aspergillus niger and Alternalia solani. Results:Methanolic extract of A. heterophyllum showed significant (P≤0.05) antifungal activity against both the tested organisms. It was also observed that ethanolic extracts of P. bistorta also had good antifungal activity against the tested fungal strains as compared to the methanolic extracts. It showed significant antifungal activity (P≤0.05) against both the tested strains. Antioxidant activity of methanolic and ethanolic extracts of A. heterophyllum and P. bistorta were also measured using a radical scavenging method. Ascorbic acid was used as a standard. Conclusions:It was observed that A. heterophyllum and P. bistorta have significant antioxidant activity. Higher antioxidant activity was recorded in methanolic extract of A. heterophyllum as compared to its ethanolic extract. However, in case of P. bistorta ethanolic extract of the plant exhibited higher antioxidant potential than methanolic extracts. Hence both of these plants have significant antimicrobial as well as antioxidant potential.