A preliminary disease survey in the wild Nile crocodile (Crocodylus niloticus) population in the Okavango Delta, Botswana.

The objective of this study was to conduct a preliminary survey of diseases that might be present in the wild Nile crocodile population in the Okavango Delta, Botswana. Blood samples were collected from crocodiles ranging in size from 34.0 cm to 463.0 cm total length. Samples were examined for blood parasites and underwent a haematological analysis. Before release the crocodiles were examined for various clinical abnormalities. Of the 144 crocodiles examined, none were visibly sick or displayed any signs of disease. No antibodies to Mycoplasma crocodyli were detected. Hepatozoon pettiti was present in 55.3% of blood smears examined, but there was no significant difference in any of the haematological values between the infected and uninfected crocodiles, and a high prevalence of Hepatozoon infection is not uncommon in other species. Only 7.6% of the examined crocodiles were infested with leeches. Further research is required for several of the crocodilian diseases, in particular to elucidate the role of wild crocodilians as reservoirs of infection.

Normal intestinal flora of wild Nile crocodiles (Crocodylus niloticus) in the Okavango Delta, Botswana.

Bacterial and fungal cultures were performed from cloacal swabs collected from 29 wild Nile crocodiles, captured in the Okavango Delta, Botswana. Sixteen species of bacteria and 6 fungal species were cultured. Individual crocodiles yielded 1-4 bacterial species, and 0-2 fungal species. The most commonly isolated bacteria were Microbacterium, Enterococcus faecalis, Aeromonas hydrophila, and Escherichia coli. No salmonellae were cultured. The most commonly occurring fungus was Cladosporium. Several of the bacterial and fungal species isolated have been implicated in cases of septicaemia in crocodilians. Knowledge of the normal intestinal flora will contribute towards the development of a crocodile-specific probiotic for use in farmed crocodiles.

Effective T-cell responses select human immunodeficiency virus mutants and slow disease progression.

The possession of some HLA class I molecules is associated with delayed progression to AIDS. The mechanism behind this beneficial effect is unclear. We tested the idea that cytotoxic T-cell responses restricted by advantageous HLA class I molecules impose stronger selection pressures than those restricted by other HLA class I alleles. As a measure of the selection pressure imposed by HLA class I alleles, we determined the extent of HLA class I-associated epitope variation in a cohort of European human immunodeficiency virus (HIV)-positive individuals (n=84). We validated our findings in a second, distinct cohort of African patients (n=516). We found that key HIV epitopes restricted by advantageous HLA molecules (B27, B57, and B51 in European patients and B5703, B5801, and B8101 in African patients) were more frequently mutated in individuals bearing the restricting HLA than in those who lacked the restricting HLA class I molecule. HLA alleles associated with clinical benefit restricted certain epitopes for which the consensus peptides were frequently recognized by the immune response despite the circulating virus's being highly polymorphic. We found a significant inverse correlation between the HLA-associated hazard of disease progression and the mean HLA-associated prevalence of mutations within epitopes (P=0.028; R2=0.34). We conclude that beneficial HLA class I alleles impose strong selection at key epitopes. This is revealed by the frequent association between effective T-cell responses and circulating viral escape mutants and the rarity of these variants in patients who lack these favorable HLA class I molecules, suggesting a significant pressure to revert.

Normal haematology and blood biochemistry of wild Nile crocodiles (Crocodylus niloticus) in the Okavango Delta, Botswana.

Wild Nile crocodiles (Crocodylus niloticus) of various size classes were captured in the Okavango Delta, Botswana. Blood was collected from the post occipital sinus and used for the determination of a wide range of haematological and biochemical parameters. These values were compared between the sexes and between 3 size classes. The values were also compared with the limited data available from farmed Nile crocodiles, as well as from other wild Nile crocodiles. The Okavango crocodiles were comparatively anaemic, and had comparatively low total protein and blood glucose levels. There was a high prevalence of Hepatozoon pettiti infection, however, there was no significant difference in haematological values between the infected and uninfected crocodiles. The values reported here will be useful in diagnostic investigations in both zoo and farmed Nile crocodiles.

Gastric nematodes of nile crocodiles, Crocodylus niloticus Laurenti, 1768, in the Okavango River, Botswana.

The ascaridoid nematodes Dujardinascaris madagascariensis Chabaud & Caballero, 1966, Dujardinascaris dujardini (Travassos, 1920), Gedoelstascaris vandenbrandeni (Baylis, 1929) Sprent, 1978 and Multicaecum agile (Wedl, 1861) Baylis, 1923 were recovered from the stomach contents of Crocodylus niloticus Laurenti, 1768 from the Okavango River, Botswana, together with Eustrongylides sp., a dioctophymatoid nematode usually parasitizing piscivorous birds. Dujardinascaris madagascariensis was present in most of the infected hosts, while the remaining species were mostly represented in single collections in one to three hosts. All four ascaridoid nematodes represent new geographic records.

Immune escape precedes breakthrough human immunodeficiency virus type 1 viremia and broadening of the cytotoxic T-lymphocyte response in an HLA-B27-positive long-term-nonprogressing child.

The emergence of cytotoxic T-lymphocyte (CTL) escape mutations in human immunodeficiency virus type 1 (HIV-1) proteins has been anecdotally associated with progression to AIDS, but it has been difficult to determine whether viral mutation is the cause or the result of increased viral replication. Here we describe a perinatally HIV-infected child who maintained a plasma viral load of <400 copies/ml for almost a decade until a nonbinding escape mutation emerged within the immunodominant CTL epitope. The child subsequently experienced a reemergence of HIV-1 viremia accompanied by a marked increase in the number of CTL epitopes targeted. This temporal pattern suggests that CD8 escape can play a causal role in the loss of immune control.

HIV evolution: CTL escape mutation and reversion after transmission.

Within-patient HIV evolution reflects the strong selection pressure driving viral escape from cytotoxic T-lymphocyte (CTL) recognition. Whether this intrapatient accumulation of escape mutations translates into HIV evolution at the population level has not been evaluated. We studied over 300 patients drawn from the B- and C-clade epidemics, focusing on human leukocyte antigen (HLA) alleles HLA-B57 and HLA-B5801, which are associated with long-term HIV control and are therefore likely to exert strong selection pressure on the virus. The CTL response dominating acute infection in HLA-B57/5801-positive subjects drove positive selection of an escape mutation that reverted to wild-type after transmission to HLA-B57/5801-negative individuals. A second escape mutation within the epitope, by contrast, was maintained after transmission. These data show that the process of accumulation of escape mutations within HIV is not inevitable. Complex epitope- and residue-specific selection forces, including CTL-mediated positive selection pressure and virus-mediated purifying selection, operate in tandem to shape HIV evolution at the population level.

The effects of anti-androgenic and estrogenic disrupting contaminants on breeding gland (nuptial pad) morphology, plasma testosterone levels, and plasma vitellogenin levels in male Xenopus laevis (African clawed frog).

Nuptial pads are areas, digits, and forearm characterized by epidermal keratin hooks and dermal breeding glands opening on the epidermis surface in male frogs. The presence of the epidermal hooks and secretory activity of the breeding glands are androgen-dependent and considered functioning during mating. Chemical pollutants released into the aquatic environment by humans are suspected of disrupting the normal hormonal pathways and functioning. Among the best known of these endocrine-disrupting contaminants (EDCs) are compounds that mimic the steroid 17beta-estradiol. However, recently specific anti-androgenic activity by certain EDCs, including DDE and several fungicides, have been shown in mammalian studies. The African clawed frog, Xenopus laevis, has been proposed as an ideal species to be used as a model system for screening of endocrine disruptors. In this study we employed a newly developed ELISA for the measurement of plasma vitellogenin (VTG), normally produced in the liver of females under the influence of estrogens but also in males when exposed to exogenous estrogens. Furthermore, we investigated the potential of breeding (nuptial) gland activity to be employed as a biomarker system for screening for anti-androgenic activity by certain EDCs. Our results indicate that the pharmaceutic anti-androgen flutamide did significantly (p < 0.05) effect the androgen-dependent breeding (nuptial) glands and plasma testosterone concentrations in male X. laevis. Our results further confirm that the dicarboximide fungicide vinclozolin mimics the anti-androgenic action of flutamide. Vinclozolin, however, did not significantly effect the plasma testosterone concentration. Results also confirm the ability of 17beta-estradiol to stimulate production and release of VTG in the liver of male X. laevis, although we could not confirm the in vivo induction of VTG by estrogenic mimics octylphenol and nonylphenol. Although more dose-response research is needed, this preliminary study confirms the potential of VTG production and breeding gland activity as biomarkers to screen chemicals and environmental samples for estrogenic and anti-androgenic activity using X. laevis males.

Inflammatory activity as an indicator of water quality: the use of human whole blood cultures.

A rapid whole blood culture (WBC) assay system was developed to monitor the inflammatory potential of water samples collected in the Western Cape, South Africa. Water contaminated with inflammatory substances induced the pro-inflammatory hormone interleukin 6 (IL-6). All water samples collected from the Eerste River, Stellenbosch, induced IL-6 secretion, and the quantity of IL-6 secreted is dependent on the concentration and origin of the sample. The lowest IL-6 inducing activity for river water was obtained for samples collected near the origin of the river. Samples at subsequent points downstream showed an increase in IL-6 inducing activity. Drinking water samples collected from selected towns in the Western Cape showed that there were major differences between the inflammatory potential of the water. Of the 15 samples assayed, 7 had low inflammatory activity, 4 had an intermediate inflammatory activity and 4 had high inflammatory activity. The water sources that have a high inflammatory activity may pose a health risk to consumers.

Osmoregulation of the Nile crocodile, Crocodylus niloticus, in Lake St. Lucia, Kwazulu/Natal, South Africa.

Nile crocodiles of three age classes, hatched in captivity and reared in fresh water, when exposed acutely to water of 17 and 35 ppt NaCl, suffered marked dehydration, were lethargic, ceased to feed and lost mass. When exposed to gradually increasing salinities (3-35 ppt), with a short acclimation period at each salinity, crocodiles survived, continued to feed and increased in mass and size. All age classes had a relatively constant plasma osmolality across the salinity spectrum. Cloacal urine osmolality varied throughout the acclimation experiment, but did not increase with increasing salinity. No significant increase was found in plasma concentrations of any of the osmolytes. There was a trend of decreasing cloacal urine [Na(+)] and [Cl(-)] and increasing cloacal urine [K(+)] with increased salinity, indicating that urine was not an important route for Na(+) and Cl(-) excretion. Crocodiles exposed to saline conditions maintained relatively constant plasma uric acid concentrations, but urinary uric acid concentrations increased markedly with increasing salinities. This suggests that uric acid is the main constituent of nitrogenous waste excretion in saline exposed Nile crocodiles. As in Crocodylus porosus, C.niloticus has the physiological ability to survive and thrive in periodically hyper-osmotic environments. However, its euryhalinity is restricted, in that acute exposure to sea water leads to dehydration, but with an acclimation period at lower salinities, it survives and thrives in sea water.

Audit of neonatal intensive care transport--closing the loop.

To audit the effectiveness of changes in transport arrangements, data on babies ventilated during transfer into a neonatal unit were compared between two periods. During the first period, August 1991-February 1993, an ad hoc transport team operated. Transport practice was changed in 1993 by forming a nine-person nursing transport team, improving training and upgrading monitoring. The second audit period was January 1994-July 1995. The groups were not significantly different for birthweight, gestation or levels of ventilation. Physiological variables were assessed with a "transport score". Improved scores for temperature and pH were achieved on completion of transfer in 1994-95 compared to 1991-93. Stabilizing prior to transfer took longer in the 1994-95 period. No serious deteriorations occurred in transit in the 1994-95 period, three in 1991-93. Audit facilitates identification of problems in transport. Staff, education and equipment changes were associated with improved audited outcomes.

Arginine-lysine relationship in rapeseed meal.

A series of experiments was conducted using White Leghorn male chicks to study the arginine-lysine relationship in rapeseed meal (RSM). Semi-purified diets containing 15% protein either from RSM or soybean meal (SBM) were fed. Arginine supplementation completely reversed the growth depression caused by the addition of lysine to RSM diets. Potassium acetate was only partially effective in alleviating the growth depression caused by supplemental lysine, while polyvinyl pyrrolidone significantly reduced chick performance. Tannic acid addition (1.5%) to SBM diets resulted in a severe growth depression which was partially corrected by the supplementation of arginine in the presence of methionine. Supplementation of these amino acids to RSM had a less drastic effect. It is suggested that a portion of the arginine in rapeseed meal may be required for metabolic processes involved in tannic acid excretion.