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.

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.

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.

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.

The effect of acid drinking water on rumen protozoa in the blesbok (Damaliscus dorcas phillipsi).

Rumen contents were collected from ten adult female blesbok, five from a mine area with only acid drinking water available and five from a control group consuming normal, non-polluted drinking water. The mean concentration of total protozoa in the normal water group was almost double that in the acid drinking water group, 24.9 x 10(3) versus 14.7 x 10(3). Percent of Entodinium was higher and Diplodinium lower in those animals drinking the acid water. The number of different protozoa species present in animals from both locations was fairly similar. Diplodinium bubalidis, Ostracodinium gracile and Diplodinium consors were present in the highest percentage in the normal water group, 18.8, 18.4 and 17.7 %, respectively. The same three species, plus Entodinium dubardi, were also highest in the acid water group, O. gracile, 21.3 %; D. consors, 12.6 %; E. dubardi, 11.4 % and D. bubalidis, 10.3 %. Seventeen species of protozoa found in this study were a new host record for the blesbok, bringing the total number of species reported from the blesbok to 29.

Capture of farmed Nile crocodiles (Crocodylus niloticus): comparison of physiological parameters after manual capture and after capture with electrical stunning.

The electric stunner (e-stunner) is commonly used to handle Nile crocodiles (Crocodylus niloticus) on commercial farms in South Africa, but while it seems to improve handling and safety for the keepers, no information regarding physiological reactions to e-stunning is currently available. The aim of this study was therefore to compare various physiological parameters in farmed C niloticus captured either manually (noosing) or by using an e-stunner. A total of 45 crocodiles were captured at a South African farm by either e-stunning or noosing, and blood samples were taken immediately as well as four hours after capture. Parameters monitored were serum corticosterone, lactate, glucose, as well as alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase and creatine kinase. Lactate concentrations were significantly higher in noosed compared with e-stunned animals (P<0.001). No other blood parameter differed significantly between the two methods of capture. In addition, recorded capture time confirmed that noosing takes significantly longer time compared with e-stunning (P<0.001), overall indicating that e-stunning seems to be the better option for restraint of especially large numbers of crocodiles in a commercial setup because it is quicker, safer and did not cause a significant increase in any of the parameters measured.

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.

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.

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.

Three case studies involving Leptospira interrogans serovar pomona infection in mixed farming units.

Three case studies involving Leptospira interrogans serovar pomona outbreaks within mixed farming systems in South Africa are described. On 2 farms, pigs constituted the main enterprise with cattle and sheep of secondary importance. On each of these 2 farms, abortion due to L. pomona in sows was confirmed by culture, and antibody titres to pomona were detected in cattle, sheep, horses and dogs. On the 3rd farm, a piggery was of secondary importance to cattle farming. Abortion and death in cows occurred on this farm and serology showed titres to various serovars, including pomona. L. pomona was also isolated from bovine urine, an aborted bovine foetus and kidneys from slaughtered pigs. This particular case study was regarded as clinically atypical in that adult Jersey cattle died of acute leptospirosis in a semiarid region of South Africa. In all 3 case studies, the poor management of pig effluent and of the drinking water and its sources played a pivotal role in the transmission of the disease. Inadequate vaccination of animals against Leptospira and poor record-keeping within the secondary farming enterprises were also contributing factors to the spread of leptospirosis.

Serological survey to determine the prevalence of bovine leukaemia virus antibodies in dairy cattle on selected farms in the Gauteng and Mpumalanga provinces.

Cattle from a farm where enzootic bovine leukosis had been diagnosed were tested to determine the prevalence of bovine leukaemia virus antibodies. Farmers who had bought cattle from this farm were identified and their herds also tested. Of 381 adult dairy cattle tested, 14 animals reacted positively (3.67%). Cattle (n = 81) from 3 selected herds, not associated with the affected farm were also bled and 7 animals reacted positively (8.64%).

A nervous disorder in cattle, caused by the plants Ficus ingens var. ingens and Ficus cordata subsp. salicifolia.

Two outbreaks of neurotoxicoses are reported in cattle browsing on the leaves of Ficus spp. In the first outbreak, three animals died and one became ill. A sheep developed severe nervous signs, including tetanic spasms, when dosed with the leaves of Ficus ingens var. ingens from the toxic camp where the cattle had died. The second outbreak resulted in the death of 12 heifers within 48 h of ingestion of the leaves of F. cordata subsp. salicifolia. Clinical signs included hyperaesthesia, ataxia, muscle tremors and padding motions while in lateral recumbency. Similar signs were reproduced by drenching the incriminated leaves to a steer. The sheep dosed with F. ingens var. ingens and two cattle, one of which had died during the second outbreak and the steer drenched with F. cordata subsp. salicifolia, were necropsied. Light microscopical examination consistently revealed oedema of the central nervous system. In the steer, focal demyelination was evident in localized areas of the brain and spinal cord. Liver lesions ranged from mild degeneration to focal disseminated necrosis of hepatocytes.

Serological survey for canine leptospirosis in the Pretoria area.

Canine serum samples (n = 400) from the Pretoria area were tested for Leptospira antibodies, using the microscopic agglutination test. The prevalence of antibodies (inconclusive and positive titres) was 1.5%. Reactions were only against Leptospira interrogans serovars tarassovi and pyrogens. Leptospirosis does not appear to be an important canine disease in the Pretoria area.

A reduction in the duration of infection with Tritrichomonas foetus following vaccination in heifers and the failure to demonstrate a curative effect in infected bulls.

Seven batches of 25% water-phase, oil-in-water vaccine were prepared from whole cultures of Tritrichomonas foetus. Two inoculations were given, spaced 6 weeks apart, to virgin heifers and infected bulls. A significant reduction (P less than 0.01) in the duration of infection in vaccinated heifers was seen when they were challenged by being bred to a bull infected with the same isolate as that contained in the vaccine. Only 1/12 vaccinated heifers were pregnant 4.5 months after the end of the breeding season compared to 2/12 in the control group. The vaccine, therefore, has no practical advantage. Vaccine was supplied to 2,724 bulls on properties where the infection was present. From these bulls, 110 reliable results were obtained, where bulls had been infected, been inoculated and tested 1 month later. No curative effect was demonstrable with 69/110 (62.7%) bulls, remaining infected after the course of inoculations. There was also no difference between vaccine batches or between bulls of different ages. Further work on improving the vaccine is indicated. Three media suitable for the culture of T. foetus are described in detail.

Serological reactions to Leptospira species in buffalo (Syncerus caffer) from the Kruger National Park.

Four hundred and six serum samples from buffalo (Syncerus caffer) were tested for leptospirosis, using the microscopic agglutination test. Seven buffaloes (1.7%) reacted positive and 27 (6.6%) inconclusive. Reactions against L. tarassovi and L. hardjo were the most prevalent.

Serological survey for bovine leptospirosis in the Volksrust district.

Serum samples (n = 860) from cattle in the Volksrust district were tested for Leptospira antibody titres. Seventeen (2%) of the animals were positive for leptospirosis, while 9(1%) animals showed suspect reactions. Titres against L. hardjo, L. pomona, and L. tarassovi were the most prevalent.

A serological survey for brucellosis in buffalo (Syncerus caffer) in the Kruger National Park.

A serological investigation was undertaken to determine the prevalence of brucellosis titres in buffalo in the Kruger National park. A total of 406 samples were collected over a period of one year. The rose bengal and the complement fixation tests were used in the investigation as these tests are routinely used for cattle sera and have proved to be reliable. In the females, 12.6% adult, 10.7% sub-adult and 3% juvenile animals reacted positively to the tests. In the males, 15.1% adults, 10.6% sub-adults and 5.3% juveniles were recorded as positive.