Mental distress, food insecurity and university student dropout during the COVID-19 pandemic in 2020: evidence from South Africa.

Background: Student dropout has been a key issue facing universities for many years. The COVID-19 pandemic was expected to exacerbate these trends; however, international literature has produced conflicting findings. Limited literature from Africa has investigated the impact of COVID-19 on student dropout trends, despite the documented devastation, including increased risk of food insecurity and mental distress, caused by the pandemic. Objective: This work seeks to understand the impact of food insecurity and mental distress on student dropout during the COVID-19 pandemic. Methods: Using a cross-sectional research design, first-year undergraduate students from a large South African university were recruited via email to participate in a survey between September and October 2020. The Household Food Insecurity Access Scale (HFIAS) was used to measure food insecurity and the Patient Health Questionnaire Anxiety and Depression Scale (PHQ-ADS) was used to measure mental distress. Multivariate regression was used to investigate factors associated with student dropout. Results: The student dropout rate was 10.5% (95% CI: 8.2-13.2). The prevalence of severe food insecurity was 25.7% (95% CI: 22.3-29.4) and the prevalence of severe mental distress symptoms was 26.7% (95% CI: 23.3-30.4). Dropout rates and levels of food insecurity were highest among students residing in remote areas during the lockdown at 19.2% and 43.6%, respectively. The multivariate logistic regression revealed that being male increased the probability of dropout almost three-fold (odds ratio (OR) = 2.70; 95% CI: 1.48-4.89, p =0.001)). Being moderately food insecure increased the odds of dropout more than two-fold (OR=2.50; 95% CI:1.12-5.55, p=0.025), and experiencing severe mental distress symptoms increased the odds of dropout seven-fold (OR=7.08; 95% CI:2.67-18.81, p<0.001). Conclusion: While acknowledging that various factors and complexities contribute to student dropout, the increased vulnerability to food insecurity and mental distress, stemming from issues such as widespread job losses and isolation experienced during the pandemic, may have also had an impact on dropout. This work reiterates the importance of directing additional support to students who are food insecure and those who are experiencing mental distress in order to mitigate university student dropout.

Failure of diplodiatoxin to induce diplodiosis in juvenile goats.

Diplodiosis is an important neuromycotoxicosis of ruminants in South Africa when grazing on harvested maize fields in winter. It is believed to be caused by mycotoxin(s) synthesised by Stenocarpella (Diplodia) maydis. Although several metabolites have been isolated from S. maydis culture material, none of these have been administered to ruminants to reproduce the disease. The objectives of this study were to isolate diplodiatoxin and to administer it to juvenile goats. Diplodiatoxin, considered as a major metabolite, was purified from S. maydis-infected maize cultures (Coligny 2007 isolate). Following intravenous administration of 2 mg and 4 mg diplodiatoxin/kg body weight for five consecutive days to two juvenile goats, no clinical signs reminiscent of diplodiosis were observed. Based on previous experimental results and if diplodiatoxin was the causative compound, the dosage regimen employed was seemingly appropriate to induce diplodiosis. In addition, intraruminal administration of 2 mg/kg diplodiatoxin to one goat for three consecutive days also did not induce clinical signs. It appears as if diplodiatoxin alone is not the causative compound. Other metabolites and/or mixtures of diplodiatoxin and other mycotoxins, when available in sufficient quantities, should also be evaluated.

Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12).

Geigeria poisoning in sheep, locally known as 'vermeersiekte', is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated muscle lesions in small stock. Because of ethical issues surrounding the use of live animals in toxicity studies, there is currently a dire need to establish an in vitro model that can be used to replace traditional animal experimentation. The objective of this study was to determine the cytotoxicity of geigerin in a murine myoblast cell line (C2C12) using methyl-thiazol-tetrazolium (MTT) and lactate dehydrogenase (LDH) assays, annexin V and propidium iodide (PI) flow cytometry and transmission electron microscopy (TEM). Mouse myoblasts were exposed to 2.0 mM, 2.5 mM and 5.0 mM geigerin for 24, 48 and 72 h. A concentration-dependent cytotoxic response was observed. Apoptosis was detected by means of annexin V flow cytometry during the first 24 h and apoptotic bodies were also visible on TEM. According to the LDH and PI flow cytometry results, myoblast cell membranes were not injured. We concluded that the murine myoblast cell line (C2C12) is a suitable model for future studies planned to evaluate the cytotoxicity of other and combinations of sesquiterpene lactones, with and without metabolic activation, implicated in 'vermeersiekte' and to elucidate the subcellular effects of these myotoxins on cultured myoblasts.

Characterization of cell death caused by diplodiatoxin and dipmatol, toxic metabolites of Stenocarpella maydis.

Diplodiosis, a neuromycotoxicosis of cattle and sheep grazing on mouldy cobs infected by Stenocarpella maydis, is considered the last major veterinary mycotoxicosis for which the causative mycotoxin is still unknown. The current study was aimed at characterizing the cell death observed in mouse neuroblastoma (Neuro-2a), Chinese hamster ovary (CHO-K1) and Madin-Darby bovine kidney (MDBK) cell lines exposed to the S. maydis metabolites (i.e. diplodiatoxin and dipmatol) by investigating the roles of necrosis and apoptosis. Necrosis was investigated using the lactate dehydrogenase (LDH) leakage and propidium iodide (PI) flow cytometry assays and apoptosis was evaluated using the caspase-3/7 and Annexin V flow cytometry assays. In addition, transmission electron microscopy (TEM) was used to correlate the cell death pathways observed in this study with their typical morphologies. Both diplodiatoxin and dipmatol (750 µM) induced necrosis and caspase-dependent apoptosis in Neuro-2a, CHO-K1 and MDBK cells. Ultrastructurally, the two mycotoxins induced mitochondrial damage, cytoplasmic vacuolation and nuclear fragmentation in the three cell lines. These findings have laid a foundation for future studies aimed at elucidating in detail the mechanism of action of the S. maydis metabolites.

Cytotoxicity of diplodiatoxin, dipmatol and diplonine, metabolites synthesized by Stenocarpella maydis.

The cytotoxicity of three Stenocarpella maydis metabolites (diplodiatoxin, dipmatol and diplonine) was investigated on Neuro-2a, CHO-K1 and MDBK cell lines. Diplodiatoxin was the most cytotoxic followed by dipmatol. Conversely, diplonine was not cytotoxic. Diplodiatoxin and dipmatol affected mitochondrial succinate dehydrogenase (MTT assay) and the overall viability of cells as assessed in real-time (xCELLigence assay). The results obtained so far indicate that diplodiatoxin and dipmatol exert their toxicity possibly via the necrotic cell death pathway.

Assessment of microcystis bloom toxicity associated with wildlife mortality in the Kruger National Park, South Africa.

Based on previous necropsy results, Microcystis blooms in constructed water impoundments in the Kruger National Park (KNP) have been identified as a cause of wildlife mortality. In response to wildlife mortality during 2007, water samples, containing algal bloom material, were collected during February 2007 and July 2007 from four dams (Nhlanganzwani, Mpanamana, Makhohlola, and Sunset) in the southeastern part of the KNP as part of the follow-up investigation. The toxicity of the Microcystis blooms was determined using the enzyme-linked immunosorbent assay (ELISA), protein phosphatase inhibition (PPI) assay, mouse bioassay, and African sharptooth catfish (Clarias gariepinus) primary hepatocytes. Both the ELISA and PPI assays indicated that the water sample collected during February 2007 from the Nhlanganzwani Dam, and samples collected from the Nhlanganzwani and Sunset dams in June 2007, were toxic. These dams, exhibiting the toxic Microcystis blooms, were also associated with the wildlife mortality. Mice injected intraperitoneally with water samples from Nhlanganzwani Dam (February 2007) induced hepatotoxicity and mortality within 1 hr. Primary hepatocytes from the sharptooth catfish exposed to samples from these dams gave similar results. This laboratory investigation and results strongly incriminate the toxic Microcystis blooms as the cause of the wildlife mortality. Eutrophication and bloom formation appear to have been the consequence of the high numbers of hippopotami (Hippopotamus amphibius) in specific dams.

A comparison of in vivo and in vitro assays to assess the toxicity of algal blooms.

The toxicity of purified microcystin-LR (MC-LR) and algal material collected during the winter and summer seasons (2005/2006) from the Hartebeespoort dam, South Africa, was investigated using the enzyme-linked immunosorbent assay (ELISA), mouse bioassay, catfish primary hepatocytes (in vitro assay) and protein phosphatase inhibition (PPi) assays. Microcystis aeruginosa, known producer of microcystins, was the dominant cyanobacteria present in the water samples. Exceptionally high cell numbers per millilitre were observed, especially with the summer samples (approximately 1.442 x 10(8)cells/ml), indicating a severe algal bloom in the dam. The toxin concentration as detected by ELISA and PPi assay in the winter and summer extracts was at least 1000 times more than the provisional guideline value (1 microg/l) set by the World Health Organization (WHO) for MC-LR in drinking water. Hepatotoxic effects and death of mice were observed after dosing with the summer extracts, while no hepatotoxic effects were observed with winter extracts. The EC(50) values obtained after exposure of the catfish primary hepatocytes for 72h to MC-LR, winter and summer extracts was about 0.091, 0.053 and 0.014 mg/l, respectively. Similar toxicity results were obtained when the mouse bioassay and primary hepatocytes were used.