Scavenger activity in a peri-urban agricultural setting in the Highveld of South Africa.

Scavenging animals often scatter skeletal remains of forensic interest and cause scavenging damage. This study aimed to identify scavenging animals in the peri-urban agricultural Highveld of South Africa, describe their scattering patterns, and the damage they cause to bone. Ten pig carcasses (Sus scrofa domesticus) (40-80 kg) were placed at the University of Pretoria's Mierjie Le Roux Experimental Farm (Highveld) in summer and winter. Motion-activated cameras recorded the scavenging. Scavenger species were identified and their behaviors, scattering pattern, and the damage they cause to bone were described. Scavenging was primarily by black-backed jackals; however, mongooses (slender, yellow, and water mongoose), Cape porcupine, and honey badger were also active. Remains were commonly scattered in two directions by jackals. The distance of scattering was heavily influenced by fencing. The remains were scattered within a maximum radius of 73.7 m. The remains were scavenged and skeletonized faster in summer. Jackals caused minimal damage to bone, isolated to superficial, nonspecific scores, furrows, and punctures. A few mongoose bone alterations were present as jagged gnaw marks on the angle of the mandible and gnawing of the vertebral spinous process. Cape porcupine bone damage included gnaw marks on the condyle of a femur and head of humerus, and destruction of the proximal and distal ends of a tibia. The described scattering pattern and bone modification patterns will assist in the recovery and analysis of scavenged remains found in peri-urban agricultural areas in South Africa.

Taphonomic bone trauma caused by Southern African scavengers.

Forensic anthropologists in Southern Africa are often confronted with post-mortem modifications of human skeletal remains caused by animal scavenging. This is troublesome as the post-mortem pseudo-trauma could be misinterpreted. This study aimed to describe the skeletal trauma caused by Southern African scavengers which are of forensic interest. The scavenging animals selected for this study included wild dog, spotted hyena, lion, leopard, black-backed jackal, caracal, and porcupine housed at the National Zoological Gardens of South Africa. Sparsely fleshed and articulated bovine front and hind limbs as well as ribs were placed in each animal enclosure and collected after 2 days for cleaning and analysis. Felids (lion, leopard, and caracal) gnawed away the greater trochanter of the humerus leaving deep, parallel furrows. Hyena caused massive trauma to bone with one-third of the tibia shaft surviving with jagged fracture edges. Porcupines left distinctive fan-like parallel scores and large, oval depressions with an eroded, polished appearance. Wild dogs and jackals did not leave any distinctive patterns. Light scavenging trauma is distinct from other bone taphonomy but cannot be used to determine the species. Heavy scavenging trauma patterns can be used to determine the general type of perpetrating animal; however, the exact taxa or species cannot be determined.

A comparison between decomposition rates of buried and surface remains in a temperate region of South Africa.

Several studies have been conducted on decomposition patterns and rates of surface remains; however, much less are known about this process for buried remains. Understanding the process of decomposition in buried remains is extremely important and aids in criminal investigations, especially when attempting to estimate the post mortem interval (PMI). The aim of this study was to compare the rates of decomposition between buried and surface remains. For this purpose, 25 pigs (Sus scrofa; 45-80 kg) were buried and excavated at different post mortem intervals (7, 14, 33, 92, and 183 days). The observed total body scores were then compared to those of surface remains decomposing at the same location. Stages of decomposition were scored according to separate categories for different anatomical regions based on standardised methods. Variation in the degree of decomposition was considerable especially with the buried 7-day interval pigs that displayed different degrees of discolouration in the lower abdomen and trunk. At 14 and 33 days, buried pigs displayed features commonly associated with the early stages of decomposition, but with less variation. A state of advanced decomposition was reached where little change was observed in the next ±90-183 days after interment. Although the patterns of decomposition for buried and surface remains were very similar, the rates differed considerably. Based on the observations made in this study, guidelines for the estimation of PMI are proposed. This pertains to buried remains found at a depth of approximately 0.75 m in the Central Highveld of South Africa.

Histomorphology, ultrastructure and fatty acid composition of the adipose tissue in pansteatitis, the potentials in understanding the underlying mechanism and diagnosis of pansteatitis in the Nile crocodile.

BACKGROUND: In an effort to characterize the fat body and other adipose tissue in the Nile crocodile and the effects of pansteatitis on the structure and composition of the adipose tissue, we evaluated the regional variation in structure and fatty acid composition of healthy farmed crocodiles and those affected by pansteatitis. METHODS: Adipose tissue samples were collected from the subcutaneous, visceral and intramuscular fat and the abdominal fat body of ten 4-year old juvenile crocodiles from Izinthaba Crocodile Farm, Pretoria, South Africa while pansteatitis samples were collected from visceral and intramuscular fat of crocodiles that had died of pansteatitis at the Olifant River, Mpumalanga, also in South Africa. Histomorphology, ultrastrustucture and fatty acid composition by fatty acid methyl ester (FAME) analysis were conducted. RESULTS: Histological examination showed regional variations in the adipose tissue especially in the collagen content of the ECM, tissue perfusion and division into lobes and lobules by fibrous capsule. Considerable fibrosis, mononuclear cell infiltration especially by macrophages and lymphocytes and toxic changes in the nucleus were observed in the pansteatitis samples. Regional variation in lipid composition especially in Myristoleic (C14:1), Erucic acid (C22:1n9), and Docosadienoic acid (C22:2n6) was observed. Most of the saturated and trans fatty acids were found in significant quantities in the pansteatitis samples, but had very low levels of the cis fatty acid and the essential fatty acids with C18 backbone. CONCLUSION: This study demonstrates that there exists some regional variation in histomorphology and fatty acid composition in the healthy adipose tissue of the Nile crocodile. It also showed that pansteatitis in the Nile crocodile might have been triggered by sudden change in energy balance from consumption of dead fish; and probable exposure to toxic environmental conditions with the evidence of up scaled monounsaturated long chain fatty acids composition and toxic changes in the leucocytes observed in pansteatitis in the present study.

Pansteatitis in polluted Olifants River impoundments: nutritional perspectives on fish in a eutrophic lake, Lake Loskop, South Africa.

This study compares the aetiology of pansteatitis in Lake Loskop, relative to two other impoundments along the Olifants River. Macroscopic and microscopic pathology, age determination and analysis of stomach content, fatty acids and stable isotopes explain the high prevalence of pansteatitis in Oreochromis mossambicus (Peters) and several other species in Lake Loskop. All the dietary indicator comparisons between pansteatitis-affected and healthy fish fail to support a systemic cause. Pansteatitis in Lake Loskop was linked to size and weight of O. mossambicus, but not to ontogenic age. Fish in Lake Loskop showed abnormally high omega-3 to omega-6 fatty acid ratios normally only found in marine fish with no significant difference in degree of assimilation of these fatty acids between pansteatitis-affected and healthy fish. This explains the vulnerability to, but not the occurrence of, pansteatitis. As a cause for the pansteatitis, these results point towards sporadic vitamin E-depleting trigger events, known sporadic fish die-off occurrences that provide surviving fish with a rich source of rancid fats on which to scavenge. The mechanism ties pansteatitis to eutrophication and trophic cascade effects, the intrinsic drivers of the disease and suggests an adaptive management strategy that might be applied by relevant conservation authorities.

Age estimation from the acetabulum in South African black males.

Anthropologists are constantly seeking to improve methods for age estimation in the human skeleton. A new method was introduced about a decade ago that assesses the morphological changes that take place in the acetabulum as an individual ages. The pelvis is usually well preserved in forensic cases, which makes this method potentially valuable as an adult age indicator. This method employs seven variables, each with its own set of phases. To test the accuracy and reliability of this method, 100 black South African male acetabula from the Pretoria Bone Collection were assessed based on the criteria described in the original study. Box plots and transition curves were constructed to establish whether progression with age was visible and how it could possibly be modelled. Inter-observer reliability was also assessed by making use of Fleiss's Kappa statistic. Five specimens were used as out-of-sample examples for which maximum likelihood (point) estimates were calculated. The results demonstrated that middle and older individuals' age estimates were vastly underestimated. Inter-observer repeatability was poor, which suggested that the classification system most likely needs to be modified. A discussion and recommendation is given for improvement of reliability and repeatability of this method.

Fluid resuscitation in acute medicine: what is the current situation?

The administration of intravenous fluids for resuscitation is the most common intervention in acute medicine. There is increasing evidence that the type of fluid may directly affect patient-centred outcomes. There is a lack of evidence that colloids confer clinical benefit over crystalloids and they may be associated with harm. Hydroxyethyl starch preparations are associated with increased mortality and use of renal replacement therapy in critically ill patients, particularly those with sepsis; albumin is associated with increased mortality in patients with severe traumatic brain injury. Crystalloids, such as saline or balanced salt solutions, are increasingly recommended as first-line resuscitation fluids for the majority of patients with hypovolaemia. There is emerging evidence that saline may be associated with adverse outcomes due to the development of hyperchloraemic metabolic acidosis, although the safety of balanced salt solutions has not been established. Fluid requirements vary over the course of critical illness. The excessive use of fluids during the resuscitative period is associated with increased cumulative fluid balance and adverse outcomes in critically ill patients. The selection of fluid depends on the clinical context in which it is administered and requires careful consideration of the dose and potential for toxicity. There is an urgent need to conduct further high-quality randomized controlled trials of currently available fluid therapy in patients with critical illness.

Choice of fluid in acute illness: what should be given? An international consensus.

Fluid management during critical illness is a dynamic process that may be conceptualized as occurring in four phases: rescue, optimization, stabilization, and de-escalation (mobilization). The selection and administration of resuscitation fluids is one component of this complex physiological sequence directed at restoring depleted intravascular volume. Presently, the selection of i.v. fluid is usually dictated more by local practice patterns than by evidence. The debate on fluid choice has primarily focused on evaluating outcome differences between 'crystalloids vs colloids'. More recently, however, there is interest in examining outcome differences based on the chloride content of crystalloid solutions. New insights into the conventional Starling model of microvascular fluid exchange may explain that the efficacy of colloids in restoring and maintaining depleted intravascular volume is only moderately better than crystalloids. A number of investigator-initiated, high-quality, randomized controlled trials have demonstrated that modest improvements in short-term physiological endpoints with colloids have not translated into better patient-centred outcomes. In addition, there is substantial evidence that certain types of fluids may independently worsen patient-centred outcomes. These include hydroxyethyl starch and albumin solutions in selected patient populations. There is no evidence to support the use of other colloids. The use of balanced salt solutions in preference to 0.9% saline is supported by the absence of harm in large observational studies. However, there is no compelling randomized trial-based evidence demonstrating improved clinical outcomes with the use of balanced salt solutions compared with 0.9% saline at this time.

Liver moisture content in animals and possible causes of variations in hepatic dry matter content.

The concentration of trace elements in the liver is used as an indicator of the mineral nutritional status of an animal, as a benchmark of environmental mineral exposure, to follow the metabolism of an element in the body and for various other purposes. Concentrations are expressed on a wet (fresh) liver basis or on a dry liver basis. From a literature search and evidence from an analytical laboratory, large variations (varying from < 20% to > 40%) have been recorded on the percentage of moisture in the livers of ruminants. Such variations potentially compromise the interpretation of results on mineral concentrations in livers, and preclude robust comparisons between studies. Among the factors that can affect the moisture content of livers are: inconsistencies in sampling and preparation of liver samples; exposure to toxic substances; ill-health of the animal; fat content of the liver; and age of the animal. It was estimated that the mean dry matter (DM) content of the livers of healthy ungulates containing less than 1% liver fat is between 27.5% and 28.5%, and on a fat-free basis 25-26% DM. For routine analyses of liver samples it is suggested that to limit variations owing to differences in liver moisture content, liver mineral concentrations should be expressed on a DM basis, and for in-depth scientific studies on mineral metabolism on a dry, fat-free basis. However, if mineral concentrations are expressed on a wet basis, it is advisable to supply the liver DM content as well.

Electroencephalogram (EEG) assessment of brain activity before and after electrical stunning in the Nile crocodile (Crocodylus niloticus).

Electrical stunning is used to capture crocodiles to perform routine management procedures. It is essential from a welfare point that electrical stunning must cause unconsciousness in animals. However, there is no information of whether or not electrical stunning causes unconsciousness in the Nile crocodile (Crocodylus niloticus). The purpose of the study was to assess brain activity before and after electrical stunning in crocodiles using a 5-channel referential electroencephalogram analysis to determine consciousness. Behavioural indicators and electroencephalogram recordings of 15 captive-bred crocodiles were captured and analysed using power spectral density analysis immediately before and after stunning and then at 60 s intervals until 5 min post-stunning. A standardised stun of 170 Volts was applied for 5-7 s on the wetted neck. Unconsciousness was defined as a decrease in alpha wave power and increase in delta wave power. Three of the electroencephalograms could not be assessed. Unconsciousness was identified in 6 out of 12 crocodiles and lasted for an average for 120 s. An increase in electroencephalogram waveform amplitude and tonic-clonic seizure-like waveform activity and behaviour indicators were not reliable indicators of unconsciousness. Further research should be focused on improving the efficiency and reliability of electrical stunning.

Geochemistry of mineral licks at Loskop Dam Nature Reserve, Mpumalanga, South Africa.

Although numerous hypotheses have been proposed to explain geophagy, the primary driver of this behaviour remains elusive. Supplementation of scarce nutrients is one commonly cited explanation. We examined the element concentration of three licks relative to adjacent topsoils to infer the possible reasons for geophagy at Loskop Dam Nature Reserve. Lick samples had greater concentrations of B, Co, Zn, Se, Mo and Mn (Loskop Main Lick); Cu (Klopperskloof Lick); and Na (Klopperskloof Lick and Rhenosterhoek Lick) than those of adjacent topsoil. We suggest that supplementation with all or some of these nutrients is a likely driver of geophagy in this fenced reserve, with different licks providing herbivores with different suites of nutrients.

Treatment rationale for dogs poisoned with aldicarb (carbamate pesticide).

The treatment rationale for dogs poisoned by aldicarb is reviewed from a pharmacological perspective. The illegal use of aldicarb to maliciously poison dogs is a major problem in some parts of the world. In South Africa, it is probably the most common canine poisoning treated by companion animal veterinarians. Aldicarb poisoning is an emergency and veterinarians need to be able to diagnose it and start with effective treatment immediately to ensure a reasonable prognosis. Successful treatment depends on the timely use of an anti-muscarinic drug (e.g. atropine). Additional supportive treatment options, including fluid therapy, diphenhydramine, benzodiazepines and the prevention of further absorption (activated charcoal) should also be considered. Possible complications after treatment are also briefly discussed.

Animal scavenging on pig cadavers in the Lowveld of South Africa.

Scavenging animals often scatter skeletal remains of forensic interest and leave bite marks. This study aimed to identify scavenging animals in the rural Lowveld of South Africa and to describe their scattering pattern and bite marks on bone. Ten pig cadavers (Sus scrofa domesticus) (40-80 kg) were placed at the Wits Rural Facility, Limpopo, South Africa during the summer and winter seasons. Motion activated cameras recorded the scavenging. Scavenger species were identified and their behaviors, scattering pattern, and bite marks were described. Scavenging was primarily by vultures (hooded, white-backed, and lappet-faced). Marabou stork, slender and banded mongoose, genet, civet, warthog and honey badger also actively scavenged. Vultures began to scavenge the pig cadavers after 18hrs in summer and between 26 and 28 h in winter and skeletonized pig cadavers rapidly between 5 and 98 min. Skeletonization occurred more rapidly and diffusely in summer while winter cases were densely scattered. Overall the scattered remains were within an area of 157.9 m2/1705.5 ft2 with a radius of 7.09 m/23.3 ft. Vultures cleaned bones thoroughly with very minimal markings - primarily nonspecific scores. The described scattering pattern and bite marks will assist in the recovery and analysis of scavenged remains.

Causative pathogens and antibiotic resistance in community-acquired urinary tract infections in central South Africa.

BACKGROUND: Urinary tract infections (UTIs) are very common in community practice. Both the South African (SA) antibiotic stewardship programme (2015) and the Essential Medicines List for SA (2018) recommend ciprofloxacin as first-line treatment for community-acquired urinary tract infections (CAUTIs). The pathogens responsible for CAUTIs and their susceptibility profiles need to be documented, which is important for developing and updating treatment protocols. OBJECTIVES: To determine the causative pathogens of CAUTIs in the greater Bloemfontein area, central SA, and to review their susceptibilities to commonly prescribed antibiotics. METHODS: Urine samples sent for microscopy and culture between 2011 and 2015 by the three largest primary healthcare facilities in Bloemfontein were analysed retrospectively. Specimens with a significant count (>105 CFU/mL) of a single uropathogen were included. These results were obtained from the National Health Laboratory Service central data warehouse after the required consent. Data regarding age, gender, pathogen cultured and antimicrobial susceptibilities were captured. All calculations were carried out with statistical analysis software SPSS 17.0 (SPSS Inc., USA). RESULTS: A total of 712 samples met the inclusion criteria. Women accounted for 481 (67.6%) of the infections. The prevalence of UTIs per age group was as follows: 1 month - 25 years (n=146; 20.51%); 26 - 50 years (n=324; 45.5%); and 51 - 75 years (n=199; 27.9%). The distribution of pathogens did not differ between age groups. Escherichia coli was the most prevalent uropathogen cultured from 410 (57.6%) specimens, followed by Klebsiella spp. from 97 (13.6%) and Enterococcus spp. from 71 (10.0%) specimens. E. coli showed resistance rates of 77.1% to amoxicillin, 15.6% to amoxicillin-clavulanate, 18.5% to ciprofloxacin, 4% to nitrofurantoin and 11% to trimethoprim-sulfamethoxazole (TMP-SMX). The distribution of uropathogens was different for men and women, with a lower prevalence of E. coli in men (p=0.045). CONCLUSIONS: As expected, E. coli comprised most of the isolates, with a higher than expected number of Klebsiella isolates cultured. The susceptibility of E. coli to commonly prescribed oral antibiotics has decreased in the research setting, which mirrors a global trend. This study provides data showing that TMP-SMX and nitrofurantoin can be used safely as alternatives to first-line ciprofloxacin in CAUTIs in central SA.

Responses of phytoplankton upon exposure to a mixture of acid mine drainage and high levels of nutrient pollution in Lake Loskop, South Africa.

The relationships between water quality and the phytoplankton community within Lake Loskop were studied during the late summer and autumn of 2008 to evaluate the impacts of acid mine drainage and high nutrient concentrations. The higher concentrations of metal ions and sulphate had adverse effects on certain phytoplankton species in the inflowing riverine zone of Lake Loskop, in comparison to the reference site in the lacustrine zone of the lake, which was dominated by the larger and slower growing late summer species of Coelastrum reticulum Nägeli, Straurastrum anatinum Meyen ex Ralfs and Ceratium hirundinella Müller. The high nutrient concentrations (nitrogen: 17 mg l(-1) and orthophosphate: 0.7 mg l(-1)) during the mid-summer peak of the rainy season were associated with the development of a bloom of the cyanobacterium Microcystis. Water quality data associated with the development of the Microcystis bloom suggest that the aquatic system of Lake Loskop has now entered an alternate, hypertrophic regime. This change overshadowed the adverse effects of high concentrations of heavy metal ions and low pH. Throughout this study, the reference site in the lacustrine zone of Lake Loskop had lower concentrations of metal ions and sulphate, and higher pH values. The response of phytoplankton bioassays on integrated water samples from the different sampling sites did provide potential answers to the reasons for the absence of the algal group Chlorophyceae in the phytoplankton community structure in the riverine zone of the lake.

Molasses as a possible cause of an "endocrine disruptive syndrome" in calves.

During the mid 1990s a potentially serious, chronic syndrome was reported in well-managed beef and dairy herds from unrelated parts of South Africa. Farmers reported that it manifested as various combinations of decreased production, decreased weaning masses, apparent immune breakdown in previously immunocompetent animals, increased reproductive disorders, various mineral imbalances in non-deficient areas and goitre, noticeable as enlarged thyroid glands. The farmers associated this syndrome with certain batches of sugar cane molasses and molasses-based products. The syndrome was reminiscent of an "endocrine disruptive syndrome". The objective of this study was to evaluate the suspected endocrine disruptive effect of molasses included in cattle feed. Using existing in vitro assays, four batches of molasses syrup were screened for possible inclusion in a calf feeding trial. Two batches were selected for the trial. Thirty-two, 4- to 6-week-old, weaned Holstein bull calves were included in the single phase, three treatment, parallel design experiment. In two of the groups of calves, two different batches of molasses were included in their rations respectively. The control group was fed a ration to which no molasses was added, but which was balanced for energy and mineral content. The mass gain of the calves was recorded over the 6-month study period. The calves were clinically examined every week and clinical pathology parameters, immune responses and endocrine effects were regularly evaluated. Even though endocrine disrupting effects were detected with the in vitro screening assays, these could not be reproduced in the calves in the experiment. The two batches of molasses utilized in the calf feeding trial did not induce major differences in any of the parameters measured, with the exception of a lower mass gain in one of the molasses-fed groups (Group 1), which tended towards significance. The results of the study indicate that the two batches of molasses had no endocrine disruptive or immunosuppressive effects in calves.

Provisional clinical chemistry parameters in the African sharptooth catfish (Clarias gariepinus).

Pollution affects aquatic systems worldwide and there is an urgent need for efficient monitoring. Fish are generally sensitive to their environment and are thus considered to be valuable bioindicator species. The African Sharptooth catfish (Clarias gariepinus) is particularly important in this respect because of its very wide distribution. In order to use C. gariepinus as a bioindicator species its baseline clinical chemistry must be defined. Existing data are scarce, and the objective of this work was therefore to establish clinical chemistry parameters for C. gariepinus. Blood was collected from male and female catfish and a number of clinical chemistry parameters were determined. Plasma protein values, but particularly those of plasma albumin, were found to be very low, approximately half the value for dogs, but similar to the values in Channel catfish (Ictalurus punctatus). Plasma urea values in Sharptooth catfish were found to be much lower than in dogs, but only marginally lower than in Channel catfish. Plasma creatinine in Sharptooth catfish, however, was only a quarter of that of dogs and one third of that found in Channel catfish. These findings may have implications for using urea and/or creatinine as an index of renal glomerular filtration, as is done in mammals. Plasma enzyme activity ranges were much lower in Sharptooth catfish than in dogs, particularly for alkaline phosphatase (ALP) and alanine aminotransferase (ALT). By comparison, Channel catfish have an even lower ALT activity range but an ALP range that is very similar to dogs. The implications for using these enzymes as markers for liver disease are not clear from these data, as factors such as plasma half-life and tissue distribution remain to be determined. The very low plasma thyroxine (T4) levels have important implications for laboratory personnel, who will have to set up calibration and standardisation adaptations for the methods that are generally designed for human samples. Although the sample size was too small for reliable comparisons, it appeared that there was little difference in the parameters measured between male and female fish. The values obtained are a useful starting point for using C. gariepinus as a bioindicator species.

Vitamin D status and supplementation in adult patients receiving extracorporeal membrane oxygenation.

The prevalence of vitamin D deficiency in critical illness is known to be high and associated with adverse clinical outcomes. Patients receiving extracorporeal membrane oxygenation (ECMO) may be at increased risk of vitamin D deficiency due to high severity of acute illness. Challenges with drug dosing in ECMO patients are recognised due to increased volume of distribution and drug absorption to circuit components. To describe the prevalence of vitamin D deficiency in ECMO patients and the effect of intramuscular dosing of cholecalciferol on levels of vitamin D metabolites, and to compare these data with intensive care unit (ICU) patients not receiving ECMO, two prospective studies were performed sequentially: an observational study of 100 consecutive ICU patients and an interventional study assessing effects of intramuscular cholecalciferol in 50 ICU patients. The subgroup of patients who required ECMO support in each of these studies was analysed and compared to patients who did not receive ECMO. Twenty-four ECMO patients, 12 from the observational study and 12 from the interventional study (who received intramuscular cholecalciferol) were studied-21/24 (88%) ECMO patients were vitamin D deficient at baseline compared to 65/126 (52%) of non-ECMO patients (P=0.006). Of the 12 ECMO patients who received cholecalciferol, six patients (50%) achieved correction of deficiency compared to 36/38 (95%) non-ECMO patients (P=0.001). The prevalence of vitamin D deficiency is higher in ECMO patients compared to other critically ill adults. Correction of deficiency with single dose cholecalciferol is not reliable; higher or repeated doses should be considered to correct deficiency.

Establishment and validation of primary hepatocytes of the African sharptooth catfish (Clarias gariepinus).

In vitro systems such as primary cells and continuous cell lines are gaining momentum in ecotoxicological studies. Cytotoxicity tests with fish cells as well as tests using specific endpoints such as CYP1A induction are valuable in the toxicity assessment of environmental samples. The main objective of this study was to establish and validate the use of primary hepatocytes from the African sharptooth catfish (Clarias gariepinus) as an in vitro toxicity monitoring system. The successful isolation of primary hepatocytes from the sharptooth catfish was achieved using an in situ perfusion method. The primary hepatocytes responded to CYP1A induction, while a continuous Chinese hamster ovary (CHO-K1) cell line showed no activity when exposed to various concentrations of benzo[a]pyrene (B[a]P) (p<0.0001). Cytotoxicity, as measured by the methyl thiazol tetrazolium (MTT) assay, was not observed following a 72 h exposure of the primary hepatocytes and the CHO-K1 cell line to different B[a]P concentrations. However, the hepatocytes were damaged at higher B[a]P concentrations (>10(-6)M) as shown by transmission electron microscopy. This cytotoxicity effect was also confirmed by the trypan blue exclusion assay (TD(50) of 10(-6)M). Differences in the results between the MTT and trypan blue exclusion assays are probably due to mitochondria that are still metabolically active, causing the tetrazolium salt to be dehydrogenated. The internal architecture of normal primary hepatocytes included large quantities of rough endoplasmic reticulum (often in close proximity to the nucleus), mitochondria, aggregates and scattered glycogen, a few lipid droplets and spherical nuclei with distinct nucleoli. The primary catfish hepatocyte cell culture system, expressing CYP1A when exposed to B[a]P, could be used as a biomarker for aromatic hydrocarbon pollutants in aquatic ecosystems of southern and East Africa.