Hatching success and survival of fish early life stages in a chronic exposure to nevirapine: a case study of the Mozambique tilapia.

The anti-retroviral nevirapine has been detected in surface waters throughout South Africa and its effects on non-target aquatic animals are still unknown. The aim was to investigate the potential effects of nevirapine on the hatching success and survival of Oreochromis mossambicus early life stages through a chronic exposure. The exposer started with newly fertilized O. mossambicus eggs and concluded 30 days after hatching. Environmental relevant concentration of nevirapine (1.48 µg/l) was used in a static renewal system and a controlled environment (27 ± 1°C; 14:10 day/night cycle). The main endpoints assessed included hatching success and survival; a morphological assessment was also done on whole individual on day 1 and 30 post-hatching to identify any physical abnormality. Nevirapine had no noticeable effects on the hatching success and survival of O. mossambicus larvae; no statistically significant differences were observed between the control and the nevirapine exposed fish (p > 0.05).

The effects of environmentally relevant concentrations of aldrin and methoxychlor on the testes and sperm of male Clarias gariepinus (Burchell, 1822) after short-term exposure.

The organochlorine pesticides aldrin (0.14 µg/L) and methoxychlor (0.23 µg/L) were both present in the Albasini Dam, Limpopo Province, South Africa, during a field survey in 2014. The use of aldrin has been banned in the USA since 1987 and restricted in South Africa since 1992. The use of methoxychlor, however, remains undefined with little information available about its registration in South Africa despite being banned in Europe (2002) and USA (2003). The aim of this study was to determine the potential effects of environmentally relevant concentrations of aldrin and methoxychlor on the reproductive system of male catfish, Clarias gariepinus. Males were exposed for 96 h to the two pesticides under controlled laboratory conditions. Following exposure, each fish was weighed and measured, and a necropsy performed to determine any macroscopic abnormalities and the general health of the fish. The fish were killed and dissected and the testes removed, weighed and measured to determine the gonadosomatic index (GSI). The right testis of each fish was sectioned for histopathological assessment and to calculate the testes index (IT). The left testis was used for computer-assisted sperm analysis (CASA). The histopathological assessment of the testes showed histopathological changes such as of melano-macrophage centres (MMCs) and vacuolation of spermatogonia and spermatocytes. However, the classification of these changes indicated that the testes tissue structure was normal with slight histological changes. No statistically significant differences (p > 0.05) were found in the CASA parameters between exposure groups. The results of this study showed that the environmentally relevant concentrations of aldrin and methoxychlor did not have a negative effect on the motility of the mature sperm, but adverse effects were noted in the early stages of spermatogenesis, indicating possible effects over longer exposure periods.

The potential effects of efavirenz on Oreochromis mossambicus after acute exposure.

Antiretroviral drugs (ARVs) are hazardous therapeutic pharmaceuticals present in South African surface water. Efavirenz is an ARV commonly used in human immunodeficiency virus (HIV) treatment in South Africa. Although little is known about the toxic effects of efavirenz on fish health, threats of toxicity to the aquatic environment have been reported. Oreochromis mossambicus were exposed under controlled conditions to environmentally-relevant efavirenz concentrations (10.3ng/l) as measured in rivers that flow into the Nandoni Dam in the Vhembe District, South Africa. Acute (96h) exposures were conducted using efavirenz concentrations of 10.3ng/l and 20.6ng/l. The overall health of exposed fish was determined using a histology-based fish health assessment index. Necropsies and haematology were conducted and somatic indices calculated after which the liver, kidney, heart, gills and gonads were microscopically quantitatively assessed. Results indicated that fish exposed to 20.6ng/l efavirenz had significantly (p<0.02) higher liver indices than the control fish, indicating increased liver damage including steatosis and frank necrosis. Fish exposed to 20.6ng/l efavirenz presented with significantly (p<0.02) higher total fish indices, representative of declined overall health compared to control fish. It was concluded that the exposure of O. mossambicus to efavirenz resulted in liver damage and overall decline in fish health. These novel findings may indicate a health risk for O. mossambicus and other biota exposed to efavirenz in aquatic ecosystems. Thus, ARV's in water sources of South Africa pose a definite threat to wildlife and ultimately human health.

Organochlorine pesticide levels in Clarias gariepinus from polluted freshwater impoundments in South Africa and associated human health risks.

There are increasing concerns regarding the safe human consumption of fish from polluted, freshwater impoundments. The aim of this study was to analyse the muscle tissue of the sharptooth catfish Clarias gariepinus for selected organo-chlorine pesticides (OCPs) and to perform a human health risk assessment using a standard protocol described by the United States Environmental Protection Agency (US EPA). Fish were collected from the polluted Roodeplaat-(RDPD), Rietvlei-(RVD) and Hartbeespoort (HBPD) Dam impoundments located in the north-eastern regions of South Africa. GC-MS analyses showed levels of various OCPs in fish muscle samples from all three impoundments. For fish collected from the RDPD, p,p'-DDE, endosulfan, lindane and ß- and δ-HCH were the most prevalent OCPs detected, while p,p'-DDE and endosulfan were the most predominant in fish from the RVD. Lindane and ß- and δ-HCH were the main OCPs detected in fish from the HBPD. Dieldrin was the only OCP detected at concentrations for which a cancer risk and a hazard index above the acceptable risk levels were estimated. This was the case for fish from both the RDPD and RVD impoundments. No toxic risk was estimated should fish from the HBPD be consumed.

Intersex in feral indigenous freshwater Oreochromis mossambicus, from various parts in the Luvuvhu River, Limpopo Province, South Africa.

This study reports on intersex in Oreochromis mossambicus, an indigenous fish species inhabiting most aquatic systems throughout South Africa (SA). Male fish were collected from three sites in the Luvuvhu River, Limpopo Province, SA: Albasini Dam (AD), Nandoni Dam (ND), and Xikundu Weir (XW). The latter two sites are situated in a currently dichloro-diphenyl-trichloroethane (DDT) sprayed area. A laboratory-bred reference group (Aq R) were included for a histological comparison. 48% of the fish at AD were intersex individuals compared with 63% at ND, and 58% at XW. The Aq R fish had no cases of intersex. o,p'- and p,p'-DDT and metabolites dichlorodiphenyldichloroethane (DDD) and -dichlorodiphenyldichloroethylene (DDE) were detected in fat samples, indicative of contamination of the aquatic environment and subsequent exposure of fish to these chemicals. Although some of the fat samples contained levels of DDTs no association could be established between intersex and chemical contaminants in fish.

DDT contamination from indoor residual spraying for malaria control.

The insecticide DDT is still used in specific areas of South Africa for indoor residual spray (IRS) to control malaria vectors. Local residents could be exposed to residues of DDT through various pathways including indoor air, dust, soil, food and water. The aims of this study were to determine the levels of DDT contamination, as a result of IRS, in representative homesteads, and to evaluate the possible routes of human exposure. Two villages, exposed (DV) and reference (TV) were selected. Sampling was done two months after the IRS process was completed. Twelve homesteads were selected in DV and nine in TV. Human serum, indoor air, floor dust, outside soil, potable water, leafy vegetables, and chicken samples (muscle, fat and liver) were collected and analyzed for both the o,p'- and p,p'-isomers of DDT, DDD and DDE. DDT was detected in all the media analyzed indicating a combination of potential dietary and non-dietary pathways of uptake. DV had the most samples with detectable levels of DDT and its metabolites, and with the exception of chicken muscle samples, DV also had higher mean levels for all the components analyzed compared to TV. Seventy-nine percent of participants from DV had serum levels of DDT (mean [summation operator]DDT 7.3microg g(-1) lipid). These residues constituted mainly of p,p'-DDD and p,p'-DDE. [summation operator]DDT levels were detected in all indoor air (mean [summation operator]DDT 3900.0 ng m(-3)) and floor dust (mean [summation operator]DDT 1200.0 microg m(-2)) samples. Levels were also detected in outside soil (mean [summation operator]DDT 25.0 microg kg(-1)) and potable water (mean [summation operator]DDT 2.0 microg L(-1)). Vegetable sample composition (mean [summation operator]DDT 43.0 microg kg(-1)) constituted mainly p,p'-DDT and p,p'-DDD. Chicken samples were highly contaminated with DDT (muscle mean [summation operator]DDT 700.0 microg kg(-1), fat mean [summation operator]DDT 240,000.0 microg kg(-1), liver mean [summation operator]DDT 1600.0 microg kg(-1)). The results of the current study raise concerns regarding the potential health effects in residents living in the immediate environment following DDT IRS.

Testicular microlithiasis and neoplastic lesions in wild eland (Tragelaphus oryx): possible effects of exposure to environmental pollutants?

The purpose of the study was to compare wildlife in the proximity and away from the sources of known industrial pollution. Macroscopic, focal, gritty areas that appeared white were observed in the testes of all 24 South African eland (Tragelaphus oryx) culled in the Rietvlei Nature Reserve (RNR; n=17) between 2001 and 2003 and Suikerbosrand Nature Reserve (SNR; n=7) in 2004. Histopathological evaluation of testes showed multiple intratubular dystrophic calcifications, focal areas of sperm stasis and interstitial chronic cell infiltrates with fibrosis. Spermatogenesis was generally impaired; a few atypical germ cells were also encountered. Sertoli cell vacuolization and sloughing of the seminiferous epithelium were evident. Adenomatous changes of the rete testis, reflective of possible chronic estrogenic exposure, were found. In testes collected from three reference eland in 2007 from the Molopo Nature Reserve (MNR) in the Kalahari/Kgalagadi Desert, except for one focal area of sperm stasis and another with microcalcification, the seminiferous epithelium as well as collecting/rete tubules were normal. Analyses of fat tissue for environmental pollutants showed that 11 out of 17 RNR eland contained a detectable estrogenic chemical p-nonylphenol (mean+/-SD: 184.8+/-24.6 microg/kg fat); no organochlorine chemicals or polychlorinated biphenyls were detected. Of the 7 SNR eland, 5 had detectable octylphenol residues (50.2+/-30.9 microg/kg fat), 3 had detectable p-nonylphenol (137.8+/-77.9 microg/kg fat), 3 had o-p'-DDT (114.9+/-31.1 microg/kg fat), 3 had p-p'-DDT (127.3+/-49.9 microg/kg(79.5+/-30.4 microg/kg fat) and 5 contained o-p'-DDE (27.7+/-9.9 microg/kg fat). One eland from the MNR contained one 70.6 microg o-p'-DDT/kg fat and another p-p'-DDE 61.3 microg/kg fat. Therefore, in eland with testicular abnormalities, significant amounts of various estrogenic chemicals were bioaccumulated in fat samples. It therefore seems likely that the lesions found in RNR and SNR were associated with the relatively high body-burden of environmental pollutants (phenols), although the possibility of systemic infections cannot be ruled out. No testicular abnormalities were found in reference eland. These findings are the first indication of mammalian wildlife being affected by environmental pollution of endocrine disrupting chemicals in South Africa.

DDT residues in water, sediment, domestic and indigenous biota from a currently DDT-sprayed area.

DDT is used for indoor residual spraying (IRS) in Limpopo Province, northern South Africa to control malaria. Through IRS, DDT may reach the outdoor environment via dust and air and from possible spillages during application. In this area the local people consume domestic chickens, wild fish or birds. Fish from the river catchment and impoundments seem to be the major source of protein intake. Water, sediment and tissue samples from two such fish species, domestic chickens and wild birds (terrestrial and aquatic) from this DDT-sprayed area were analysed for DDT and metabolite residues. The samples contained p,p'-DDT, p,p'-DDD and p,p'-DDE residues, with the latter the most ubiquitous and in the highest concentrations. These findings raise concern that both water and food may be major routes of human exposure to DDT and metabolites, thereby posing possible adverse human health implications to the local communities.

Histopathological alterations in the liver of the sharptooth catfish Clarias gariepinus from polluted aquatic systems in South Africa.

There is a need for sensitive bio-monitoring tools in toxicant impact assessment to indicate the effect of toxicants on fish health in polluted aquatic ecosystems. Histopathological assessment of fish tissue allows for early warning signs of disease and detection of long-term injury in cells, tissues, or organs. The aim of this study was to assess the degree of histopathological alterations in the liver of C. gariepinus from two dams in an urban nature reserve, (Gauteng, South Africa). Two dams (Dam 1 and Dam 2) were chosen for their suspected levels of toxicants. Water and sediments were sampled for metal and potential endocrine disrupting chemical analysis. A quantitative and qualitative histology-based health assessment protocol was employed to determine the adverse health effects in fish. The analysis of blood constituents, fish necropsy, calculation of condition factors, and hepatosomatic indices were employed to support the findings of the qualitative and quantitative histological assessment of liver tissue. Assessment of the liver tissue revealed marked histopathological alterations including: structural alterations (hepatic cord disarray) affecting 27% of field specimens; plasma alterations (granular degeneration 98% and fatty degeneration 25%) of hepatocytes; an increase in melanomacrophage centers (32%); hepatocyte nuclear alterations (90%); and necrosis of liver tissue (14%). The quantitative histological assessment indicated that livers of fish collected from Dam 1 were more affected than the fish livers collected from Dam 2.

Histological evidence of intersex in feral sharptooth catfish (Clarias gariepinus) from an estrogen-polluted water source in Gauteng, South Africa.

This is the first histological evidence of intersex in a fish species inhabiting a South African water source. One hundred catfish (Clarias gariepinus) were collected randomly from the Marais Dam (MD) and the Rietvlei Dam (RVD) in the Rietvlei Nature Reserve (RNR), South Africa. These dams drain water from a stream that receives effluent from industrial sites, agricultural activities, informal settlements, and municipal treatment plants. Each fish was evaluated macroscopically and had blood drawn, and its gonads were macroscopically and histologically examined to verify intersex potentially related to endocrine disruption. Gonadal histology of several fish showed primary oocytes scattered in testicular tissue, indicative of intersex. The results showed intersexuality in 20% of fish from both the dams. The GSI value for intersex fish was closer to male GSI values, suggesting that the sampled intersex fish were more likely to have occurred from the feminization of male catfish. Target chemical analyses showed that the water, sediment, and serum samples tested positive for p-nonylphenol (p-NP). The p-NP level in water and sediment at MD was 6360 and 4.0 microg/kg, respectively, whereas in sediment at RVD it was 113 microg/kg. Commonly found in the effluent from sewage treatment plants, p-NP in water and sediment indicates estrogenic water pollution, which might affect wildlife and humans dependent on these sources.

The use of different enzymes in feral freshwater fish as a tool for the assessment of water pollution in South Africa.

Selected enzymes in the organs/tissues of two fish species (Oreochromis mossambicus and Labeo umbratus) were evaluated as biomarkers of water pollution in warm- and cold-water systems. The fish were caught using standard techniques during two high-flow and two low-flow surveys. The results indicated that the fluctuating enzyme activities measured were not a result of pollution but could be initiated as a result of (1) no or too little pollution, (2) handling of the fish, and (3) struggling in the nets. Furthermore, most of the enzymes are nonspecific for the specific pollutants present. It is concluded that the selected enzymes could not be successfully employed as biomarkers of water pollution when the organisms are sampled from a polluted water source.