Some antibiotics have lost their efficacy over common infections and this has led to the search for new antibiotics and chemically altering existing ones for a better control of infectious diseases. In the present study, Pyrenacantha grandiflora tubers extracts were conjugated with ampicillin, penicillin, vancomycin and silver nanoparticles and their antimicrobial activity was evaluated against Escherichia coli, Staphylococcus aureus and Klebsiella Pneumoniae. The reactions were confirmed by formation of new functional groups that were identified by Fourier transmission infrared spectroscopy (FTIR). Minimum inhibitory concentrations were determined using the microdilution assay. Minimum bactericidal concentrations and the fractional inhibition concentration index were also determined. FTIR analysis indicated different functional group associated with conjugation. The activity of ampicillin was improved when conjugated with silver nanoparticles against K. pneumonia and E. coli. Vancomycin showed improvement of activity when conjugated to silver nanoparticles against K. pneumonia. Penicillin was improved by acetone extracts and vancomycin showed to be more effective when conjugated with silver nanoparticles and water extracts. The conjugation of P. grandiflora with penicillin, ampicillin and vancomycin in the presence of silver nanoparticles improved their biological activities. Therefore, the conjugates are medicinally important and can be used to improve the activity of existing antibiotics.

The implementation of water and sanitation safety plans (WSSP) has the potential of greatly improving the challenge of resource-limited drinking-water supplies. However, the most effective tool to make WSSP successful is understanding of the factors that contribute to hindering the implementation of these plans, specifically in rural communities. This study therefore aimed at assessing the status of basic services and determining the factors that contribute to hampering the process of WSSP in rural communities. A survey was conducted between March 2020 and March 2021 in rural communities of the Vhembe District, Limpopo Province, South Africa. The overall results indicate that poverty, unemployment, lack of access to purified water, and inadequate sanitation facilities have resulted in waterborne diseases reported within the communities and have a major impact in hindering WSSP. Other barriers observed are inequality regarding financial power, absent and degrading water and sanitation infrastructures, and lack of protection and maintenance of natural water sources. Therefore, there is a need for community members to be educated on proper behavior and perceptions towards sanitation, including working hand-in-hand with different stakeholders, men and women from communities, and different cultures and religions to overcome these barriers, so that human disease associated with water supply, wastewater reuse, and sanitation in rural communities can be alleviated.

The aim of the current study was to evaluate, validate and select microbial water quality monitoring tools to establish their suitability and feasibility for use in rural communities. The monitoring of water quality was performed at three different levels: i) basic level focusing on sanitary inspection and hydrogen sulphide (H₂S) test; ii) intermediate level via enumeration of faecal indicator bacteria (faecal coliforms, Escherichia coli, Enterococcus spp. and Clostridium perfringens); and iii) advanced level based on qPCR detecting host-associated genetic markers (BacHum, BacCow, Cytb, Pig-2-Bac, and BacCan) and pathogens (Vibrio cholerae, Escherichia coli O157:H7, and Shiga toxin-producing Escherichia coli). A positive correlation was recorded between sanitary risk and faecal coliforms (r = 0.613 and p < 0.002), E. coli (r = 0.589 and p < 0.003), and Enterococcus spp. (r = 0.625 and p < 0.003). The H₂S test showed positive correlations with sanitary risk score (r = 0.623; p < 0.003), faecal coliforms (r = 0.809; p < 0.001), E. coli (r = 0.779; p < 0.001) and Enterococcus spp. (r = 0.799; p < 0.001). Similar correlation patterns were also found with advanced techniques used for detecting host-associated genetic markers, excepted between Clostridium perfringens, and Pig-2-Bac (pig), BacCan (dog), and V. cholerae. The H₂S test and sanitary inspections are therefore suitable and cost-effective tools to capacitate rural areas at household level for the monitoring of faecal contamination and management of water sources.

Numerous human activities and poor sanitation management cause public health concern, particularly in rural communities without reliable water supply systems and resources for the monitoring of the quality of their water sources. This study assessed the relationship between observed sanitary risks and hydrogen sulphide (H₂S) strip test results in the identification of faecal contamination of various water sources used at household level in rural areas of the Vhembe District Municipality. The highest percentage sanitary risk scores ranging from 50 to 100% were recorded for both river and dam water commonly used by the households for multiple purposes, including drinking. All the surface water samples (100%) also tested positive for H₂S production, which is linked to the contamination of water sources by bacteria of faecal origin. The overall results showed a significant and positive correlation (r = 0.623, p = 0.003 in the wet season and r = 0.504, p = 0.017 in the dry season) between sanitary risk scores and H₂S strip test results. In low resource settings, the use of sanitary inspections combined with the inexpensive and easy-to-use H₂S strip tests can be effective as drinking water quality management tools to raise an awareness among community members of the faecal contamination of their water sources.

In the present study, silver nanoparticles (AgNPs) were synthesized using both the chemical and biological methods and conjugated with Pyrenacantha grandiflora extracts. These were then characterized and evaluated for antimicrobial activities against multi-drug resistant pathogens, such as methicillin-resistant Staphylococcus aureus (MRSA), Klebsiella pneumonia, and Escherichia coli. Nanoparticles were analyzed with UV-visible spectrophotometer, transmission electron microscopy (TEM), and energy dispersive X-ray analysis (EDX). Silver nanoparticles, P. grandiflora extracts, and the conjugates were also analyzed with Fourier transform infrared spectroscopy (FTIR). As a result, quasi-sphere-shaped AgNPs with sizes ranging from 5 to 33 nm and spherically shaped AgNPs with sizes ranging from 3 to 25 nm were formed from chemical and biological synthesis, respectively. A well diffusion assay showed that the activity of silver nanoparticles was most improved with acetone extract against all tested bacteria with diameters in the range of 19–24 mm. The lowest MIC value of 0.0063 mg/mL against MRSA was observed when biologically synthesized AgNPs were conjugated with acetone and water extracts. Chemically synthesized silver nanoparticles showed the lowest MIC value of 0.0063 mg/mL against E. coli when conjugated with acetone and methanol extracts. This study indicates that silver nanoparticles conjugated with P. gandiflora tubers extracts exhibit strong antibacterial activities against multi-drug resistant bacterial pathogens. Therefore, biosynthesized conjugates could be utilized as antimicrobial agents for effective disease management due to the synergistic antibacterial activity that was observed.

Consumption of contaminated drinking water is known to cause waterborne diseases such as diarrhoea, dysentery, typhoid, and hepatitis. This study applied whole-genome sequencing (WGS) to detect, identify, compare, and profile diarrhoeagenic pathogens (Vibrio cholerae, Shiga toxin-producing Escherichia coli, and Escherichia coli O157:H7) from 3168 water samples and 135 faecal samples (human and animal). Culture-based methods, MALDI-TOF mass spectrometry, and PCR were employed prior to WGS for identification of pathogens. Culture-based results revealed high presumptive prevalence of STEC (40.2%), V. cholerae (37.1%), and E. coli O157:H7 (22.7%). The MALDI-TOF confirmed 555 isolates with V. cholerae identified as Vibrio albensis. Shiga toxin-producing Escherichia coli (STEC) was more prevalent in wastewater (60%), treated water (54.1%), and groundwater (36.8%). PCR detected 46.4% of virulence genes from the water isolates and 66% of virulence genes from the STEC stool isolates. WGS also revealed STEC (92.9%) as the most prevalent species and found common virulence (e.g., hcp1/tssD1 and hlyE) and resistance (e.g., acrA and baeR) genes in all three types of samples. Five resistance and thirteen virulence genes overlapped among treated water and stool isolates. These findings highlight the diarrhoeagenic pathogens’ public health risk in water sources and underscore the need for better water quality monitoring and treatment standards.