Corrigendum: Hepatogenous photosensitisation in cows grazing turnips (Brassica rapa) in South Africa.

No abstract available.

Hepatogenous photosensitisation in cows grazing turnips (Brassica rapa) in South Africa.

Holstein cows on a farm in the Humansdorp district, Eastern Cape province, South Africa, developed reddened, painful teat skin 3 days after grazing a mixed forage crop dominated by bulb turnip (Brassica rapa, Barkant cultivar). The crop was grazed 45 days after planting and 10% of the herd developed symptoms. More characteristic non-pigmented skin lesions started manifesting 1-2 days after the appearance of the teat lesions. Affected cows had elevated serum activities of gamma-glutamyl transferase, glutamate dehydrogenase and aspartate aminotransferase. These blood chemistry findings confirmed a secondary (hepatogenous) photosensitivity. As a result of the severity of the teat and skin lesions, seven cows were slaughtered and tissue samples from five of them were collected for histopathological examination. Liver lesions in cows that were culled 3 or more weeks after the onset of the outbreak showed oedematous concentric fibrosis around medium-sized bile ducts and inflammatory infiltrates in portal tracts. Characteristic lesions associated with other known hepatobiliary toxicities were not found. No new cases were reported 5 days after the cattle were removed from the turnips. The sudden introduction of the cows, without any period of transitioning or adaptation to grazing turnips, as well as the short latent period, clinical signs of photosensitisation, blood chemistry and histopathology, confirmed a diagnosis of Brassica-associated liver disease, a condition seen in New Zealand but not previously described in South Africa. Brassica forage crops are potentially toxic under certain conditions and farmers must be aware of these risks.

Organochlorine pesticide bioaccumulation in wild Nile crocodile (Crocodylus niloticus) fat tissues: Environmental influences on changing residue levels and contaminant profiles.

Biologically significant concentrations of organochlorine pesticides (OCPs) continue to be reported in wildlife populations and are of particular concern in species that occupy the highest trophic levels. Nile crocodiles (Crocodylus niloticus) are important apex predators occurring throughout much of tropical and subtropical sub-Saharan Africa, where they inhabit estuarine and freshwater habitats often impacted by contamination. In this study we examined pesticide residue accumulation in fat tissue from Nile crocodiles at Lake St Lucia, South Africa, where historically large quantities of OCPs have been used for agriculture and disease control. During 2019, we collected tail fat samples from wild (n = 21) and captive (n = 3) individuals to examine the influence of habitat, body size and sex on variations in bioaccumulation. The principal contaminant found was p,p'-DDE, a major persistent metabolite of DDT, which continues to be used in the region for combating malaria. Tissue p,p'-DDE concentrations in wild crocodiles (95-1200 ng g-1 ww) were significantly (p < 0.05) higher compared to captive individuals (23-68 ng g-1 ww) and strongly correlated (R2 > 0.70) to body length. Male (n = 14) and female (n = 7) wild crocodiles exhibited similar contaminant body burdens, however, total concentrations were substantially lower than those measured in the same population during 2016/2017. Marked differences in residue levels and profiles appear to reflect changes in food availability and dietary exposure associated with a shift in environmental conditions. These findings suggest that periods of environmental stress may be associated with enhanced toxicological risk in crocodiles. Additional work is needed to better understand contaminant accumulation and elimination mechanisms in crocodiles, and their potential effects on reproductive health.

In ovo manipulation of Nile crocodile embryos: egg windowing and potential dental research applications.

Crocodilians exhibit continuous tooth replacement (i.e., polyphyodonty) and have been identified as suitable models for tooth regeneration research due to the similarity in dental cavity and tooth anatomy between these creatures and humans. Various studies reporting in ovo bird embryo manipulation exist, but such reports for reptiles are virtually non-existent. Egg windowing enables direct access to oviparous vertebrate embryos and is therefore an important component of in ovo embryo manipulation experiments. The aim of the present study was to window Nile crocodile, Crocodylus niloticus eggs and assess the potential of direct manipulations, targeting the pharyngeal region where the maxilla and mandible originates. Crocodylus niloticus eggs were successfully windowed, and a limited number of individuals survived the entire gestation period. The 1st trimester of gestation was the most sensitive, and 96.78% of the mortalities occurred within this period. Our data indicate the suitable window for embryo manipulation targeting the mandibular arch and maxillary process, without a risk of damaging the chorioallantoic membrane (CAM) (which may be fatal), was between day six and eight after laying for embryos incubated at 31°C. This data will be of use for future embryo-based experiments related to jaw and tooth development in crocodiles as well as human tooth regeneration research.

Shuni Virus in Wildlife and Nonequine Domestic Animals, South Africa.

We screened nonequine animals with unexplained neurologic signs or death in South Africa during 2010-2018 for Shuni virus (SHUV). SHUV was detected in 3.3% of wildlife, 1.1% of domestic, and 2.0% of avian species. Seropositivity was also demonstrated in wildlife. These results suggest a range of possible SHUV hosts in Africa.

Morphological changes associated with Nile crocodile (Crocodylus niloticus) phallic glans inflation.

The crocodylian phallic glans is the distal inflatable structure that makes the most direct contact with the female cloacal and associated reproductive tract openings during copulation. Therefore, its form and function directly impact female tissue sensory interactions and insemination mechanics. Compared to mammals, less is known about glans functional anatomy among other amniotes, including crocodylians. Therefore, we paired an ex vivo inflation technique with magnetic resonance imaging 3D-reconstructions and corresponding histological analyses to better characterize the morphological glans restructuring occurring in the Nile crocodile (Crocodylus niloticus) at copulation. The expansion of contiguous inflatable spongiform glans tissues is variably constrained by adjacent regions of dense irregular collagen-rich tissues. Therefore, expansion shows regional differences with greater lateral inflation than dorsal and ventral. Furthermore, this enlargement elaborates the cup-like glans lumen, dorsally reorients the glans ridge, stiffens the blunt and bifid glans tip, and putatively works to seal the ventral sulcus spermaticus semen conduit groove. We suggest how these dynamic male structures may interact with structures of the female cloacal urodeum and how these morphological changes, in concert with the varying material properties of the structural tissue compartments visualized in this study, aid copulatory gamete transfer and resulting fecundity. RESEARCH HIGHLIGHTS: Nile crocodile glans inflation produces a reproductively relevant copulatory structure directing insemination and female tissue interactions. Pairing magnetic resonance imaging 3D reconstruction with corresponding histology effectively studies functional anatomy.

West Nile Virus in Wildlife and Nonequine Domestic Animals, South Africa, 2010-2018.

West Nile virus (WNV) lineage 2 is associated with neurologic disease in horses and humans in South Africa. Surveillance in wildlife and nonequine domestic species during 2010-2018 identified WNV in 11 (1.8%) of 608 animals with severe neurologic and fatal infections, highlighting susceptible hosts and risk for WNV epizootics in Africa.

Next generation sequencing and RNA-seq characterization of adipose tissue in the Nile crocodile (Crocodylus niloticus) in South Africa: Possible mechanism(s) of pathogenesis and pathophysiology of pansteatitis.

BACKGROUND: Concerted efforts to identify the pathogenesis and mechanism(s) involved in pansteatitis, (a generalized inflammation of the adipose tissue), that was attributed to the recent crocodile die off in the Olifants River and Loskop Dam in Kruger National Park, Mpumalanga, South Africa have been in the forefront of research in recent time. As part of the efforts, molecular characterization of healthy and pansteatitis adipose tissue was carried out by RNA sequencing (RNA-Seq) using Next Generation Sequencing (NGS) and de novo assembly of the adipose transcriptome, followed by differential gene expression analysis. METHODOLOGY: Healthy adipose tissue consisting of fifty samples was collected from the subcutaneous, visceral, intermuscular adipose tissues and the abdominal fat body of ten 4 years old juvenile crocodiles from a local crocodile farm in Pretoria, South Africa. Ten pansteatitis samples were collected from visceral and intermuscular adipose tissues of five crocodiles that were dying of pansteatitis. RESULTS: Forty-two thousand, two hundred and one (42,201) transcripts were assembled, out of which 37, 835 had previously been characterized. The de novo assembled transcriptome had an N50 (average sequence) of 436 bp, percentage GC content of 43.92, which compared well with previously assembled transcripts in the saltwater crocodile. Seventy genes were differentially expressed and upregulated in pansteatitis. These included genes coding for extracellular matrix (ECM) signaling ligands, inflammatory cytokines and tumour necrosis factor alpha (TNFα) receptors, fatty acid synthase and fatty acid binding proteins, peroxisome proliferator-activated receptor gamma (PPARγ), nuclear factor and apoptosis signaling ligands, and mitogen activated protein kinase enzymes among others. Majority (88.6%) of the upregulated genes were found to be involved in hypoxia inducible pathways for activation of NFkß and inflammation, apoptosis, Toll-like receptor pathway and PPARγ. Bicaudal homologous 2 Drosophila gene (BICD2) associated with spinal and lower extremity muscle atrophy was also upregulated in pansteatitis while Sphingosine -1-phosphate phosphatase 2 (SGPP2) involved in Sphingosine -1- phosphate metabolism was downregulated. Futhermore, Doublesex-mab-related transcription factor 1 (DMRT1) responsible for sex gonad development and germ cell differentiation was also downregulated. CONCLUSION: Thus, from the present study, based on differentially expressed genes in pansteatitis, affected Nile crocodiles might have died partly due to their inability to utilize stored triglycerides as a result of inflammation induced insulin resistance, leading to starvation in the midst of plenty. Affected animals may have also suffered muscular atrophy of the lower extremities and poor fertility.

Diagnostic Value of Energy Dispersive Hand-Held X-ray Fluorescence Spectrometry in Determining Trace Element Concentrations in Ovine Liver.

There are no data available on the use of hand-held X-ray fluorescence (XRF) spectrometry to determine trace element concentrations in veterinary diagnostics. The hand-held XRF spectrometer is easy to use and does not require extensive training for the operator. In Sub-Saharan Africa with few centralised analytical laboratories equipped with expensive apparatus or mass spectrometry capabilities, trace element analysis using the hand-held XRF spectrometer provides an alternative. The objective of this study was to compare ovine hepatic copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), selenium (Se) and zinc (Zn) concentrations as obtained with the hand-held XRF spectrometer to those of a reference laboratory using inductively coupled plasma mass spectrometry (ICP-MS). Thirty ovine livers were obtained from an abattoir; prepared as wet blended and oven-dried samples and analysed. Bayesian correlation was used to assess the correspondence between results from the XRF and ICP-MS analyses. The oven-dried preparation procedure for XRF provided the best correlation with the ICP-MS data. The correlations for Cu and Zn were strong and the XRF method may represent a suitable substitute for ICP-MS analysis. For Mn and Fe the correlations were moderately strong and the XRF method may be suitable. For Mo, the correlation was weak and XRF cannot be recommended. Selenium could not be detected in samples prepared by either method. Hand-held XRF spectrometry was a practical method to determine liver concentrations of specific trace elements under African conditions and may significantly reduce the turn-around time of analysis, but unfortunately the apparatus is expensive.

Accumulation of organochlorine pesticides in fat tissue of wild Nile crocodiles (Crocodylus niloticus) from iSimangaliso Wetland Park, South Africa.

Nile crocodiles (Crocodylus niloticus) are important apex predators in many tropical and subtropical aquatic habitats throughout much of sub-Saharan Africa. In South Africa, large crocodile populations inhabit lakes and wetlands that are impacted by organochlorine pesticides (OCPs). Despite the continued use of these compounds and their potential adverse effects on key wildlife populations in southern Africa, limited ecotoxicoloigcal data exist. In this study, we examined the accumulation of OCPs in fat tissues of live, wild Nile crocodiles from iSimangaliso Wetland Park, a region of significant biological importance. All samples (n = 15) contained multiple contaminants in highly elevated concentrations, with total residue burdens varying between 3600 and 8000 ng g-1 ww. DDT and its metabolites were the dominant compounds detected in most samples, with ∑DDT concentrations ranging between 520 and 3100 ng g-1 ww. Elevated levels of other OCPs were also detected, including lindane (67-410 ng g-1 ww), aldrin (150-620 ng g-1 ww) and heptachlor (170-860 ng g-1 ww). Our findings show that crocodiles are exposed to OCPs throughout their range within iSimangaliso Wetland Park and contain some of the highest concentrations ever recorded in crocodilian tissue. Results indicate the need for a greater understanding of the impacts of OCP exposure and toxicological responses in crocodiles from iSimangaliso, and in Nile crocodile populations in general. The novel surgical technique described in this study provides an effective method for assessing relationships between contaminant body burdens and their potential reproductive and developmental consequences in crocodilians.

Vertical transmission of microcystins to Nile crocodile (Crocodylus niloticus) eggs.

Cyanobacteria or blue green algae are known for their extensive and highly visible blooms in eutrophic, stagnant freshwater bodies. Climate change and global warming have also contributed to a rise in toxic cyanobacterial blooms. One of the most important cyanobacteria is Microcystis aeruginosa, which can synthesize various microcystins that can affect the health of terrestrial and aquatic animals. Commercial Nile crocodile (Crocodylus niloticus) farming in South Africa is based on keeping breeders (adult males and females) in big dams on farms (captive-bred approach). Unfortunately, cyanobacterial blooms in the breeder dams are a concern to farm owners, managers and veterinarians. The main objectives of this research project were to determine if microcystins were present in the contents of crocodile eggs and the liver and yolk of dead hatchlings, and to determine if the reduced hatchability on commercial farms might be caused by these toxins. Furthermore, the concentration of microcystins in the breeder dam was monitored on a monthly basis spanning the ovulation and egg laying period. During the hatching season microcystin concentrations in unfertilised eggs, egg shell membranes and in the yolk and liver of dead hatchlings were determined using liquid chromatography-high resolution mass spectrometry (LC-HRMS). Microcystins were detected in Nile crocodile egg and hatchling samples. Microcystin (MC-LR, MC-RR, MC-YR) concentrations in the crocodile egg and hatchling samples collected from clutches with a good hatching rate (≥90%) ranged between 0 and 1.76 ng g-1, with the highest concentration in the egg shell membranes. Microcystin concentrations in samples collected from clutches with a bad hatching rate (≤10%) ranged from 0 - 1.63 ng g-1 with the highest concentration detected in the hatchling yolk. However, the concentrations were probably underestimated as the percentage recovery from spiked samples was very low with the extraction method employed. Bayesian analysis suggests that the liver, yolk and unfertilised egg all have similar microcystin concentrations, while the membranes have (with moderate to high certainty) higher microcystin concentrations. There appears to be no difference in microcystin concentrations among good and bad clutches across all tissue types or within a specific tissue type, but due to the small sample size, it was not possible to determine whether microcystin affected the hatchability of Nile crocodile eggs. However, vertical transmission of microcystin variants to the Nile crocodile egg does occur and the possible implications for the survival of wild Nile crocodile populations should be ascertained.

Investigation of stillbirths, perinatal mortality and weakness in beef calves with low-selenium whole blood concentrations.

In this on-farm investigation, we report on stillbirths, weakness and perinatal mortality seen in calves on a commercial beef farm in the Roossenekal area, Mpumalanga province, South Africa. Post-mortem examination of these calves and histopathological examination of organ and tissue samples did not indicate an infectious aetiology. Affected calves had marginal to deficient whole blood selenium concentrations. Whole blood samples collected from adult cattle on this farm and five neighbouring farms were deficient in selenium. The potential contributions of other minerals to the symptoms seen are a subject of ongoing investigation, but selenium deficiency was marked in this herd and required urgent correction. Methods to correct the deficiency included the use of injectable products, and an oral selenium supplement chelated to methionine. Selenium availability to plants is primarily determined by the selenium content of the parent bedrock, the presence of other minerals and the pH of the soil. The apparent sudden onset of this problem implicates a soil factor as being responsible for reducing selenium's bioavailability in this area. Selenium deficiency can have a significant impact on human health. HIV and/or AIDS, various forms of cancer and several specific clinical syndromes are associated with selenium deficiency in humans, and the impact on human health in this area also requires further investigation.

Blood lead concentrations in free-ranging Nile crocodiles (Crocodylus niloticus) from South Africa.

Generally crocodilians have received little attention with regard to the effects of lead toxicity despite their trophic status as apex, generalist predators that utilize both aquatic and terrestrial habitats, thereby exposing them to a potentially wide range of environmental contaminants. During July-October 2010 we collected whole blood from 34 sub-adult and adult free-ranging Nile crocodiles (Crocodylus niloticus) from three separate populations in northeastern South Africa in order to analyze their blood lead concentrations (BPb). Concentrations ranged from below detectability (<3 µg/dL, n = 8) to 960 µg/dL for an adult male at the Lake St Lucia Estuary. Blood lead concentrations averaged 8.15 µg/dL (SD = 7.47) for females and 98.10 µg/dL (SD = 217.42) for males. Eighteen individuals (53 %) had elevated BPbs (≥10 µg/dL). We assessed 12 general linear models using Akaike's Information Criterion (AIC) and found no significant statistical effects among the parameters of sex, crocodile size and population sampled. On average, crocodiles had higher BPbs at Lake St Lucia than at Ndumo Game Reserve or Kosi Bay, which we attribute to lead sinker ingestion during normal gastrolith acquisition. No clinical effects of lead toxicosis were observed in these crocodiles, even though the highest concentration (960 µg/dL) we report represents the most elevated BPb recorded to date for a free-ranging vertebrate. Although we suggest adult Nile crocodiles are likely tolerant of elevated Pb body burdens, experimental studies on other crocodilian species suggest the BPb levels reported here may have harmful or fatal effects to egg development and hatchling health. In light of recent Nile crocodile nesting declines in South Africa we urge further BPb monitoring and ecotoxicology research on reproductive females and embryos.

Non-invasive assessment of adrenocortical function in captive Nile crocodiles (Crocodylus niloticus).

The occurrence of stress-inducing factors in captive crocodilians is a concern, since chronic stress can negatively affect animal health and reproduction, and hence production. Monitoring stress in wild crocodiles could also be beneficial for assessing the state of health in populations which are potentially threatened by environmental pollution. In both cases, a non-invasive approach to assess adrenocortical function as a measure of stress would be preferable, as animals are not disturbed during sample collection, and therefore sampling is feedback-free. So far, however, such a non-invasive method has not been established for any crocodilian species. As an initial step, we therefore examined the suitability of two enzyme-immunoassays, detecting faecal glucocorticoid metabolites (FGMs) with a 11ß,21-diol-20-one and 5ß-3α-ol-11-one structure, respectively, for monitoring stress-related physiological responses in captive Nile crocodiles (Crocodylus niloticus). An adrenocorticotropic hormone (ACTH) challenge was performed on 10 sub-adult crocodiles, resulting in an overall increase in serum corticosterone levels of 272% above the pre-injection levels 5h post-injection. Saline-treated control animals (n=8) showed an overall increase of 156% in serum corticosterone levels 5h post-administration. Faecal samples pre- and post-injection could be obtained from three of the six individually housed crocodiles, resulting in FGM concentrations 136-380% above pre-injection levels, always detected in the first sample collected post-treatment (7-15 days post-injection). FGM concentrations seem comparatively stable at ambient temperatures for up to 72 h post-defaecation. In conclusion, non-invasive hormone monitoring can be used for assessing adrenocortical function in captive Nile crocodiles based on FGM analysis.

Lead isotope ratios in bone ash of blesbok (Damaliscus pygargus phillipsi): a means of screening for the accumulation of contaminants from uraniferous rocks.

This study was done to determine whether blesbok (Damaliscus pygargus phillipsi) from the Krugersdorp Game Reserve (KGR) in Gauteng Province, South Africa have higher concentrations of (238)U and higher (206)Pb/(204)Pb and (207)Pb/(204)Pb ratios in their bone ash than blesbok from a nearby control reserve that is not exposed to mine water and has no outcrops of uraniferous rocks. Eight blesbok females from the KGR and seven from the control site, all killed with a brain shot, were used. A Thermo X-series 2 quadrupole ICPMS was used to measure the concentrations of (238)U and lead and a Nu Instruments NuPlasma HR MC-ICP-MS to measure the lead isotope ratios in the tibial ash from each animal. KGR blesbok had higher mean concentrations of (238)U (P = 0.02) and ratios of (206)Pb/(204)Pb and (207)Pb/(204)Pb (P < 0.00001) than the control blesbok. The probability of rejecting the false null hypothesis of no difference in the (206)Pb/(204)Pb or (207)Pb/(204)Pb ratios between KGR and control reserve animals (the power of the test) was 0.999. The blesbok from the KGR accumulated contaminants from an uraniferous environment. The (206)Pb/(204)Pb and (207)Pb/(204)Pb ratios in tibial ash proved effective in confirming accumulation of contaminants from uraniferous rocks.

The post-occipital spinal venous sinus of the Nile crocodile Crocodylus niloticus: its anatomy and use for blood sample collection and intravenous infusions.

The post-occipital sinus of the spinal vein is often used for the collection of blood samples from crocodilians. Although this sampling method has been reported for several crocodilian species, the technique and associated anatomy has not been described in detail in any crocodilian, including the Nile crocodile (Crocodylus niloticus). The anatomy of the cranial neck region was investigated macroscopically, microscopically, radiographically and by means of computed tomography. Latex was injected into the spinal vein and spinal venous sinus of crocodiles to visualise the regional vasculature. The spinal vein ran within the vertebral canal, dorsal to and closely associated with the spinal cord and changed into a venous sinus cranially in the post-occipital region. For blood collection, the spinal venous sinus was accessed through the interarcuate space between the atlas and axis (C1 and C2) by inserting a needle angled just off the perpendicular in the midline through the craniodorsal cervical skin, just cranial to the cranial borders of the first cervical osteoderms. The most convenient method of blood collection was with a syringe and hypodermic needle. In addition, the suitability of the spinal venous sinus for intravenous injections and infusions in live crocodiles was evaluated. The internal diameter of the commercial human epidural catheters used during these investigations was relatively small, resulting in very slow infusion rates. Care should be taken not to puncture the spinal cord or to lacerate the blood vessel wall using this route for blood collection or intravenous infusions.

Pansteatitis of unknown etiology associated with large-scale Nile crocodile (Crocodylus niloticus) mortality in Kruger National Park, South Africa: pathologic findings.

Annual mortality events in Nile crocodiles (Crocodylus niloticus) in the Olifants River Gorge in Kruger National Park, South Africa, were experienced between 2008 and 2012, during which at least 216 crocodiles died. Live crocodiles were lethargic. Necropsy examination of 56 affected crocodiles showed dark yellow-brown firm nodules in both somatic fat and the abdominal fat body. In all of the 11 crocodiles submitted for histology, degenerative, necrotic, and inflammatory changes supported a diagnosis of steatitis in both fat types. Crocodiles are apex predators in this anthropogenically changed aquatic ecosystem that is used by humans upstream and downstream from the park for domestic, agricultural, fishing, and recreational purposes. This pathologic review of pansteatitis in crocodiles in the Olifants River system was part of a broad multidisciplinary research program. To date, no definitive causative agent has been identified. Epidemiologic evidence suggests that this event may have been a one-time event with long-standing repercussions on the health of the crocodiles. Pathologic findings are rarely documented in wild crocodilians. This study also reports on other conditions, including the presence of coccidian oocysts, capillarid and filaroid nematodes, digenetic trematodes, and pentastomes.

Bioaccumulation of aluminium and iron in the food chain of Lake Loskop, South Africa.

Concentrations of total aluminium (Al) and iron (Fe) were determined in Lake Loskop over a period of four months in 2009 in samples of phytobenthos, phytoplankton, macroinvertebrates, amphibians and fish. The highest concentrations of Al and Fe were measured in the filamentous algae Spirogyra fluviatilis (Hillse) and Spirogyra adanata (Kütz), (Al=18,997.5mgkg(-1) dry weight and Fe=22,054.2mgkg(-1) dry weight) in the riverine zone of the lake with a near-neutral water average pH of 7.3. However, a negative correlation exists between the Al and Fe concentrations measured in the filamentous algae in comparison with the corresponding concentrations of these elements in the water column of the riverine zone. The Al concentrations in the macroinvertebrate families collected ranged from 140.6 to 385.7mgkg(-1) dry weight, with the highest values measured for Al and Fe in the family Gomphidae (385.7 and 1710.0mgkg(-1) dry weight, respectively) in comparison to other macroinvertebrate families sampled. Al and Fe concentrations (2580 and 10,697mgkg(-1) dry weight) in the stomach contents of adult Oreochromis mossambicus fishes were much higher in comparison with adult Micropterus salmoides fishes (98.5 and 439.6mgkg(-1) dry weight), respectively. In all cases of dissected fish species either white or yellow body fat was observed, thus in none of the samples both type of body fats occurred simultaneously. The concentrations of total Al and Fe in the different organs of O. mossambicus were along a mean sequence of intestine>yellow body fat>brain>gills>liver>heart>white body fat, while the mean sequence of total Al and Fe in M. salmoides was: intestine>gills>liver>heart>brain>white body fat. From the levels of Al detected in the yellow body fat of the studied fish species O. mossambicus, we suggest that this phenomenon may be related to the feeding habits of this species. Furthermore, the intake of certain species of phytobenthos by O. mossambicus could have played a role in the bioaccumulation of Al in the food chain and the possible development of pansteatitis in predators at higher trophic levels.

Technique for the collection of clear urine from the Nile crocodile (Crocodylus niloticus).

Urine samples can be a very useful diagnostic tool for the evaluation of animal health. In this article, a simple technique to collect urine from the Nile crocodile (Crocodylus niloticus) was described, based on a similar unpublished technique developed for the American alligator (Alligator mississippiensis) using a canine urinary catheter. With this technique, it was possible to collect relatively clean urine samples from Nile crocodiles of different sizes using canine urinary catheters or small diameter stomach tubes. Based on the gross anatomical features of the cloaca of the Nile crocodile, it was confirmed that urine accumulates in a chamber consisting of the urodeum and coprodeum. Faecal material is stored temporarily in the very short rectum, which is separated from the urinary chamber by the rectocoprodeal sphincter.

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.