Screening and evaluation of cytotoxicity and antiviral effects of secondary metabolites from water extracts of Bersama abyssinica against SARS-CoV-2 Delta.

BACKGROUND: Bersama abyssinica is a common herb in Africa, with diverse medical uses in different areas. The plant is well-known in Tanzania for treating respiratory disorders such as TB, tonsillitis, bronchitis, and asthma, and it has lately been utilized to treat COVID-19 symptoms. Water extract of leaf and stem bark has been registered as an herbal medication known as 'Coviba Dawa' in Tanzania for the relief of bacterial respiratory infections. The extracts, however, have not been scientifically tested for their anti-viral activities. The aim of this work was to test for the cytotoxicity and antiviral effects of bioactive ingredients from B. abyssinica extracts against the Delta variant of the SARS-CoV-2 coronavirus. METHODS: B. abyssinica leaves and stem bark were dried under shade in room temperature and then pulverized to obtain small pieces before soaking into different solvents. One hundred grams of each, leaves and stem bark, were extracted in petroleum ether, dichloromethane, ethyl acetate and methanol. Water extract was obtained by decoction of stem bark and leaves into water. Phenols, flavonoids, tannins, and antioxidants were confirmed as components of the extracts. Analysis of polar extracts of bark stem bark and leaves was done. Antiviral screening and cytotoxicity experiments were conducted in a Biosafety Level 3 (BSL-3) Laboratory facility according to International Standard Operating Procedures (SOPs). RESULTS: By the use of LC-MS/MS analysis, this study confirmed the existence of four phenolic compounds in B. abyssinica water extract; 2,4-di-tert-butylphenol, 4-formyl-2-methoxyphenyl propionate, 7,8-Dihydroxy-4-methylcoumarin, and 2,3, 6-trimethoxyflavone with antioxidant activity. This study showed that, while the water extracts of B. abyssinica had significant antiviral activity against SARS Cov2 virus, it showed no cytotoxicity effect on Vero E6 cells. In particular, the water extract (Coviba dawa) showed 75% while ethylacetate fraction of B. abyssinica leaves showed a 50% in vitro viral inhibition, indicating that these substances may be useful for the development of future anti-viral agents. CONCLUSION: We therefore recommend isolation of compounds for further profiling and development with a broader concentration range. We further recommend studies that determine the antiviral activity of extracts of B.abyssinica on other viral pathogens of clinical concern.

Assessment of raw milk quality and stakeholders' awareness on milk-borne health risks in Arusha City and Meru District, Tanzania.

A cross-sectional study was conducted to assess the quality of raw milk and stakeholders' awareness on milk-borne health risks and factors for poor milk hygiene in Arusha City and Meru District, Tanzania between October and December 2012. A total of 105 smallholder dairy farmers, milk vendors and milk retailers were interviewed, and milk samples were collected for physical, microbial and antibiotic residue analysis using standard procedures. Questionnaire results indicated high level of awareness (94 %) that drinking raw milk can predispose consumers to milk-borne diseases; nevertheless, 65 % of respondents consumed raw milk. Physicochemical analyses showed some of the milk had sediments (20 %), bad smell (21 %) and had clotted on alcohol test (27 %). About 36 % of milk samples had pH below 6.6, and 25 % had specific gravity below 1.028 g/ml. The mean total viable count (TVC) of milk from vendors is significantly (P < 0.05) higher than that from retailers and smallholder dairy farmers. Generally, 65 % of milk samples assessed had a higher TVC than the level recommended (2.0 × 10(5) cfu/ml) by the East African Community (EAC) standards. Up to 91 % of the milk samples had bacterial growth that included Eschericia coli (66 %), Staphylococcus aureus (33 %), Corynebacterium (11 %) and Pseudomonas (10 %). All smallholder dairy farmers were aware of drug residues, but majority (57 %) were unaware of human health effects caused by veterinary drug residues in milk. Up to 97 % of respondents reported to comply with drug withdrawal periods. This possibly led to all milk samples analysed to be negative from detectable levels of antibiotic residues. It is concluded that the level of awareness on milk quality is high, although practices associated with milking and post-harvest handling predispose milk to bacterial contamination which is a public health risk to milk consumers.

Effect of resinous extract from Commiphora swynnertonii (Burrt) on experimental coccidial infection in chickens.

A crude resinous extract from Commiphora swynnertonii was tested against an experimental coccidial infection in local chickens. A total of 80 growing chickens were randomly assigned into five groups, which received different treatments. Chickens in G1 were not infected with coccidian oocysts and therefore served as a negative control. All chickens in G2, G3, G4 and G5 were infected through oral administration of coccidian oocysts suspension at a dosed rate of 1.5 × 10(4) Eimeria spp. oocysts per bird. Starting from day 3 post-infection (p.i), chickens in different groups were treated for 7 consecutive days as follows: G1 and G2 (positive control) received 5 ml of normal saline as placebo, G3 and G4 were given the extract at 400 and 800 mg/kg bodyweight whereas G5 received anticoccidial drug. Clinical signs, bodyweights, oocysts counts and mortality rates were observed regularly. Results showed that oral administration of the resinous extract to chickens with coccidiosis significantly reduced mortality rate from 94 to 25 % and oocysts counts from 1.03 × 10(5) to 6.55 × 10(3) oocysts/g faeces (p < 0.05). Also a body condition score chart indicated less severe clinical signs of the disease in the groups which received the extract. Mean daily body weights were slightly reduced by the administration of the extract but this effect disappeared by day 7 p.i. These findings clearly indicate that resinous extract from C. swynnertonii has significant anticoccidial effect against experimental Eimeria spp. infection in chickens. A larger field trial to validate the use of the extract in chickens naturally infected with Eimeria spp. is required.

Antiviral activity of crude extracts from Commiphora swynnertonii against Newcastle disease virus in ovo.

Studies were carried out to investigate the effect of crude extracts from resin, leaves, stem barks and root barks of Commiphora swynnertonii against Newcastle disease virus (NDV) using an in ovo assay. Nine-day-old embryonated chicken eggs were divided into seven groups (n = 6) and received various treatments. Six groups were inoculated with velogenic NDV strain; five groups out of these were treated with different concentrations of the four extracts or a diluent, dimethylsulphoxide. The uninoculated and inoculated groups were left as negative and positive controls, respectively. Embryo survival was observed daily and embryo weights were measured day 5 post-inoculation; a few eggs from selected groups were left to hatch. Allantoic fluid from treated eggs and serum from hatched chicks were collected for hemagglutination and hemagglutination inhibition (HI) tests to detect NDV in the eggs and antibodies against NDV in the hatched chicks respectively. Results showed that embryo survival and mean embryo weight were significantly higher (p < 0.001) in those groups which were treated with the crude extracts from C. swynnertonii than the positive control group. Also the extracts significantly (p < 0.001) reduced virus titres, whereas no viruses were detected in the allantoic fluids of the resin-treated group at the highest concentration of 500 µg/mL. Furthermore, the HI test results showed very low levels of antibodies against NDV in chicks hatched from resin and root bark extract-treated eggs suggesting that these plant materials were capable of destroying the NDV before stimulating the developing chick's immunity. The current findings have clearly demonstrated that crude extracts especially that of resin from C. swynnertonii have strong antiviral activity against NDV in ovo. In vivo trials are needed to validate the use of resin from the tree in controlling Newcastle disease in chickens.