Exposure of rural households to toxic cyanobacteria in container-stored water

Cyanobacteria are potent producers of cyanotoxins that may present a health risk to people. This is especially important in rural areas where people use untreated surface water, containing cyanobacteria, for household purposes including cooking and drinking. Water is collected from these sources mainly in plastic containers, transported home and stored during use. This study investigated the occurrence of cyanobacteria and their associated toxins in these containers as well as in the associated surface water sources. The results suggest that cyanobacteria are transferred from the water sources to the containers and then survive and possibly grow in biofilm forming inside the vessels. Their associated cyanotoxins were not found in any health-significant quantities in containers. However, the occurrence of cyanobacteria in the water used by the households collected in containers clearly indicates that it can be an important route of exposure especially if toxic cyanobacteria are present in the source water. In several cases a risk of cyano-intoxication might exist unless the households undertake preventative measures.Keywords: cyanobacteria, cyanotoxin, microcystin, surface water sources, drinking water containers, biofil

A rapid and low-cost DNA extraction method for isolating Escherichia coli DNA from animal stools

The price of commercial DNA extraction methods makes the routine use of polymerase chain reaction amplification (PCR) based methods rather costly for scientists in developing countries. A guanidium thiocayante-based DNA extraction method was investigated in this study for the isolation of Escherichia coli (E. coli) DNA from goat, chicken, pig, cow and human stool samples. Two versions of the lysis buffer, with and without α-casein, were tested to alleviate PCR inhibition associated with DNA isolated from stool samples. Results obtained show that, this method using the lysis buffer containing α-casein, produces PCR ready DNA at a fraction of the cost of commercial DNA extraction kits.Key words: DNA extraction, Escherichia coli, polymerase chain reaction amplification (PCR), stool samples

A rapid and low-cost DNA extraction method for isolating Escherichia coli DNA from animal stools

The price of commercial DNA extraction methods makes the routine use of polymerase chain reaction amplification (PCR) based methods rather costly for scientists in developing countries. A guanidium thiocayante-based DNA extraction method was investigated in this study for the isolation of Escherichia coli (E. coli) DNA from goat, chicken, pig, cow and human stool samples. Two versions of the lysis buffer, with and without _-casein, were tested to alleviate PCR inhibition associated with DNA isolated from stool samples. Results obtained show that, this method using the lysis buffer containing _-casein, produces PCR ready DNA at a fraction of the cost of commercial DNA extraction kits & copy; 2011 Academic Journals

Development of new host-specific Bacteroides qPCRs for the identification of fecal contamination sources in water

Bacteroides spp. have been proposed as indicators of fecal contamination in microbial source tracking (MST) methodologies. The aim of this study was to develop new qPCR assays that target host-specific Bacteroidal 16S ribosomal RNA genes, to determine the source of fecal contamination in water. Denaturing gradient gel electrophoresis (DGGE) was used to select for host-specific bands of Bacteroides associated with a fecal pollution source and later to design four qPCR host-specific assays. A set of common primers for Bacteroides spp., four different Bacteroides spp. host-associated hydrolysis probes (human, cattle, pig, and poultry), and one hydrolysis probe for the Bacteroides genus were designed. This set of qPCR assays together with other previously developed Bacteroidetes MST targets were used to analyze water samples with fecal contamination from the four sources studied. The host-specific Bacteroides qPCRs designed for human (HMprobeBac), pig (PGprobeBac), and poultry (PLprobeBac) were highly specific for its sources (1.0, 0.97, and 1.0, respectively) although its sensitivity was lower (0.45, 0.50, and 0.73, respectively). The cattle-specific qPCR was totally unspecific and was discarded for future experiments. When compared to previously designed assays, the human and pig qPCRs showed better accuracies (0.86 and 0.84) than their counterparts HF183 and Pig-2-Bac (0.38 and 0.65). Thus, the newly designed human, pig, and poultry qPCR assays outperform other methods developed until date and may be useful for source tracking purpose

Rapid concentration and sensitive detection of hookworm ova from wastewater matrices using a real-time PCR method

The risk of human hookworm infections from land application of wastewater matrices could be high in regions with high hookworm prevalence. A rapid, sensitive and specific hookworm detection method from wastewater matrices is required in order to assess human health risks. Currently available methods used to identify hookworm ova to the species level are time consuming and lack accuracy. In this study, a real-time PCR method was developed for the rapid, sensitive and specific detection of canine hookworm (Ancylostoma caninum) ova from wastewater matrices. A. caninum was chosen because of its morphological similarity to the human hookworm (Ancylostoma duodenale and Necator americanus). The newly developed PCR method has high detection sensitivity with the ability to detect less than one A. caninum ova from 1 L of secondary treated wastewater at the mean threshold cycle (CT) values ranging from 30.1 to 34.3. The method is also able to detect four A. caninum ova from 1 L of raw wastewater and from ~4 g of treated sludge with mean CT values ranging from 35.6 to 39.8 and 39.8 to 39.9, respectively. The better detection sensitivity obtained for secondary treated wastewater compared to raw wastewater and sludge samples could be attributed to sample turbidity. The proposed method appears to be rapid, sensitive and specific compared to traditional methods and has potential to aid in the public health risk assessment associated with land application of wastewater matrices. Furthermore, the method can be adapted to detect other helminth ova of interest from wastewater matrices

Comparison of concentration methods for rapid detection of hookworm ova in wastewater matrices using quantitative PCR

Hookworm infection contributes around 700 million infections worldwide especially in developing nations due to increased use of wastewater for crop production. The effective recovery of hookworm ova from wastewater matrices is difficult due to their low concentrations and heterogeneous distribution. In this study, we compared the recovery rates of (i) four rapid hookworm ova concentration methods from municipal wastewater, and (ii) two concentration methods from sludge samples. Ancylostoma caninum ova were used as surrogate for human hookworm (Ancylostoma duodenale and Necator americanus). Known concentration of A. caninum hookworm ova were seeded into wastewater (treated and raw) and sludge samples collected from two wastewater treatment plants (WWTPs) in Brisbane and Perth, Australia. The A. caninum ova were concentrated from treated and raw wastewater samples using centrifugation (Method A), hollow fiber ultrafiltration (HFUF) (Method B), filtration (Method C) and flotation (Method D) methods. For sludge samples, flotation (Method E) and direct DNA extraction (Method F) methods were used. Among the four methods tested, filtration (Method C) method was able to recover higher concentrations of A. caninum ova consistently from treated wastewater (39-50%) and raw wastewater (7.1-12%) samples collected from both WWTPs. The remaining methods (Methods A, B and D) yielded variable recovery rate ranging from 0.2 to 40% for treated and raw wastewater samples. The recovery rates for sludge samples were poor (0.02-4.7), although, Method F (direct DNA extraction) provided 1-2 orders of magnitude higher recovery rate than Method E (flotation). Based on our results it can be concluded that the recovery rates of hookworm ova from wastewater matrices, especially sludge samples, can be poor and highly variable. Therefore, choice of concentration method is vital for the sensitive detection of hookworm ova in wastewater matrices

Learning through assessment

This book aims to contribute to the discourse of learning through assessment within a self-directed learning environment. It adds to the scholarship of assessment and self-directed learning within a face-to-face and online learning environment. As part of the NWU Self-Directed Learning Book Series, this book is devoted to scholarship in the field of self-directed learning, focusing on ongoing and envisaged assessment practices for self-directed learning through which learning within the 21st century can take place. This book acknowledges and emphasises the role of assessment as a pedagogical tool to foster self-directed learning during face-to-face and online learning situations. The way in which higher education conceptualises teaching, learning and assessment has been inevitably changed due to the COVID- 19 pandemic, and now more than ever we need learners to be self-directed in their learning. Assessment plays a key role in learning and, therefore, we have to identify innovative ways in which learning can be assessed, and which are likely to become the new norm even after the pandemic has been brought under control. The goal of this book, consisting of original research, is to assist with the paradigm shift regarding the purpose of assessment, as well as providing new ideas on assessment strategies, methods and tools appropriate to foster self-directed learning in all modes of delivery