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.

Erythrocyte morphology and haemoglobin types of neonatal roan antelopes (Hippotragus equinus) with hypochromic poikilocytic anaemia.

Neonatal, poikilocytic anaemia in some members of the Hippotragini has previously been documented but not fully investigated. This study was undertaken to describe the erythrocyte morphology of roan antelopes (Hippotragus equinus) during the first 4 weeks after birth and to identify aspects of haemoglobin (Hb) production that might be implicated in this syndrome. Twenty-nine roan antelope calves were sampled on, or close to, 1, 7, 14 and 28 days after birth. Erythrocyte morphology was characterized, and microhaematocrit values and Hb parameters determined, for each sampling occasion. Findings indicated a significant change in erythrocyte morphology during the neonatal period and two haemoglobin types, fetal and adult, were identified. The perinatal onset of adult Hb synthesis was delayed relative to the termination of fetal Hb production, resulting in the observed anaemia. Haemoglobin concentration and erythrocyte morphology were significantly correlated. These findings suggest an intimate relationship between Hb synthesis and the observed poikilocytosis. An imbalance in the synthesis of the alpha- and beta-globin chains of Hb (a thalassaemia) may prove to be the underlying pathophysiology of this syndrome.

A canine model of normovolaemic acute anaemia.

The objective was to develop a non-terminal, acute normovolaemic anaemia model in dogs that has minimal effects on patient well-being. Eleven normal Beagle dogs were used. About 20% of the circulating blood volume was removed from the jugular vein 1-3 times per day over a 3-4 day period until a haematocrit (Ht) of 13-17% was obtained. Normovolaemia was maintained by replacing the volume deficit of the red blood cells with Ringer's lactate and re-infusing the plasma. Full blood count and Ht were monitored twice daily. The 13-17% Ht was reached within 3-4 days with the number of phlebotomies ranging from four to seven. The model was primarily developed to determine echocardiographic values as well as Doppler abdominal splanchnic blood flow parameters in anaemic dogs as part of a study that will compare these results to similar studies in babesiosis-induced anaemia. The model may also be useful in the evaluation of the pathophysiology of anaemia in dogs or as a model for anaemia in humans.

Are urea and creatinine values reliable indicators of azotaemia in canine babesiosis?

Serum urea and creatinine are extensively used as parameters to screen for azotaemia. Their reciprocal plots roughly correlate with glomerular filtration rate (GFR). They are, however, subject to influence by non-renal factors and to increase their specificity they are often tested concurrently. In renal disease they are expected to behave similarly, with both parameters increasing as GFR decreases. Haemolysis, as it occurs in canine babesiosis, may cause non-renal elevations in serum urea, possibly due to ammonia loading. Furthermore, haemolysis with its related elevations in serum bilirubin and serum haemoglobin, may negatively bias the measurement of serum creatinine due to interference of these substances with the chemical analysis of serum creatinine. This negative bias occurs when the alkaline picrate method, or when direct enzymatic methods based on the measurement of hydrogen peroxide, are used. In order to investigate the significance of these perturbations in canine babesiosis, paired values of serum urea and serum creatinine from Babesia canis-negative, non-haemolysis dogs (Group 1), were used to establish a relationship between urea and creatinine over a range of azotaemia by linear regression analysis. This relationship was then used to predict serum creatinine values from actual serum urea values in B. canis-positive dogs (Group 2). The mean of the predicted serum creatinine values for Group 2 (237.03 micromol/l) was then compared with the mean of the actual serum creatinine values for Group 2 (131.31 micromol/l). For Group 2, the mean actual serum creatinine demonstrated a significant negative bias relative to the mean predicted creatinine value. There was also a higher correlation between serum urea and serum creatinine in Group 1 than in Group 2. These findings may have been caused by either nonrenal elevations of serum urea values or by interference with the measurement of serum creatinine. Therefore, although it is possible that some Group 2 dogs with B. canis with high serum urea and normal, low, or zero values for serum creatinine were not azotaemic, it is also possible that other Group 2 dogs with these biochemical findings did in fact have azotaemia. This study concluded that urea and creatinine do not behave in a similar and predictable manner over a range of azotaemia in canine babesiosis and are therefore not ideally suited for the detection of renal disease in this clinical setting.

Thrombocytopaenia in canine babesiosis and its clinical usefulness.

Canine babesiosis is a common cause of thrombocytopaenia but there are few formal studies that have investigated this haematological finding in dogs. Thrombocyte counts from full blood counts were retrospectively analysed for the years 1996-2002. Thrombocyte counts and mean platelet volumes of dogs with babesiosis were compared with those of dogs, seen over the same period of time, that did not have babesiosis. There were 1162 cases in the Babesiosis group and 10 808 in the Non-babesiosis group. A frequency distribution of the thrombocyte counts showed a trimodal distribution in the Non-babesiosis group compared to a bimodal distribution in the Babesiosis group, with a strong positive skewness. The modes for the frequency distributions were 10, 40, 300 and 10, 35 x 10(9)/l thrombocytes, respectively. The median thrombocyte count in the Babesiosis group was 14 x 10(9)/l and 282 x 10(9)/l in the Non-babesiosis group. There was a statistically significant difference in the median thrombocyte count between the Babesiosis group and the Non-babesiosis group. In the Babesiosis group, 99% of the thrombocyte counts were below the lower reference range value (250 x 10(9)/l) and 62% of thrombocyte counts were below 25 x 10(9)/l. The mean platelet volume (11.1 fl) for the Babesiosis group was greater than the reference range (6-10 fl) and significantly larger than in the Non-babesiosis group (median 9.7 fl). Thrombocyte counts greater than 110 and 250 x 10(9)/l had a predictive value that the dog was not suffering from babesiosis of 99.3% and 99.8%, respectively. There was a statistically significant difference between the thrombocyte counts of dogs with babesiosis when grouped by parasitaemia scores. The mechanisms of the thrombocytopaenia are not fully understood, and multiple mechanisms, including concomitant thrombocytopaenia-inducing diseases such as ehrlichiosis, probably result in this haematological finding. Babesiosis in the South African canine population is associated with thrombocytopaenia in nearly all patients and is severe in the majority of them. In the absence of thrombocytopaenia, babesiosis is an unlikely diagnosis.

The mixed acid-base disturbances of severe canine babesiosis.

Thirty-four dogs suffering from severe babesiosis caused by Babesia canis rossi were included in this study to evaluate acid-base imbalances with the quantitative clinical approach proposed by Stewart. All but 3 dogs were severely anemic (hematocrit <12%). Arterial pH varied from severe acidemia to alkalemia. Most animals (31 of 34; 91%) had inappropriate hypocapnia with the partial pressure of CO2 < 10 mm Hg in 12 of 34 dogs (35%). All dogs had a negative base excess (BE; mean of - 16.5 mEq/L) and it was below the lower normal limit in 25. Hypoxemia was present in 3 dogs. Most dogs (28 of 34; 82%) were hyperlactatemic. Seventy percent of dogs (23 of 33) were hypoalbuminemic. Anion gap (AG) was widely distributed, being high in 15, low in 12, and normal in 6 of the 33 dogs. The strong ion difference (SID; difference between the sodium and chloride concentrations) was low in 20 of 33 dogs, chiefly because of hyperchloremia. Dilutional acidosis was present in 23 of 34 dogs. Hypoalbuminemic alkalosis was present in all dogs. Increase in unmeasured strong anions resulted in a negative BE in all dogs. Concurrent metabolic acidosis and respiratory alkalosis was identified in 31 of 34 dogs. A high AG metabolic acidosis was present in 15 of 33 dogs. The lack of an AG increase in the remaining dogs was attributed to concurrent hypoalbuminemia, which is common in this disease. Significant contributors to BE were the SID, free water abnormalities, and AG (all with P < .01). Mixed metabolic and respiratory acid-base imbalances are common in severe canine babesiosis, and resemble imbalances described in canine endotoxemia and human malaria.

Experimental studies with Stronglyloides papillosus in goats.

Unusual clinical and pathological observations in the field in goats and sheep suffering from Strongyloides papillosus infection prompted experimental work on this parasite. Goats were infected percutaneously with either single or multiple, low or high levels of S. papillosus. Young goats up to 12 months of age were found to be the most susceptible. Some animals, however, showed substantial resistance to infective doses. Clinical signs included transient diarrhoea, misshapen, elongated faecal pellets terminally, dehydration, anorexia, cachexia, gnashing of teeth, foaming at the mouth, anaemia and nervous signs such as ataxia, a wide-based stance, stupor and nystagmus. A 'pushing syndrome' was seen in 22% of the animals. The pathological changes are described and included enteritis, status spongiosus in the brain, hepatosis leading to rupture of the liver, nephrosis, pulmonary oedema, interstitial pneumonia and pneumonia. About 6% of the goats died acutely from fatal hepatic rupture. The development of an acquired immunity was determined. The immunity elicited an allergic skin reaction at the application site of larvae or injection sites of larval metabolites. This immunity, however, could be breached by large doses of larvae. The most profound clinicopathological changes induced by the parasites were an anaemia (most pronounced in the young goats) and hypophosphataemia. Trace element analyses provided evidence of Cu, Mn and possibly Se deficiencies in some goats.

Canine babesiosis in South Africa: more than one disease. Does this serve as a model for falciparum malaria?

South African canine babesiosis is caused by the virulent Babesia canis rossi. In recent years, this common disease has been detected in 12% of dogs presented at the outpatients' division of the University of Pretoria's (Onderstepoort) Veterinary Academic Hospital, and 31% of the affected dogs have been hospitalized as seriously ill. Of these hospitalized cases, 50% had severe anaemia at presentation, 32% had moderate anaemia and 18% were non-anaemic (often polycythaemic), frequently with central-nervous-system signs or multiple organ failure. A retrospective survey of 662 hospitalized cases revealed that the haematology, clinical biochemistry and patient profile (signalment) of the severely anaemic dogs were distinct from those of the non-anaemic, indicating that the babesiosis in these two groups of dogs should be viewed as two different disease in terms of the postulated, underlying, 'pathomechanisms'. The severely anaemic dogs exhibited hypoxic hepatic disease and an increase in serum urea (without a concomitant increase in creatinine), seldom had profound electrolyte imbalances and tended to have a much more profound leucocytosis, consisting of a left-shifted inflammatory leucogram, with higher numbers of circulating metamyelocytes, lymphocytosis and monocytosis. In contrast, the non-anaemic dogs exhibited severe azotaemia (which could be of renal or pre-renal origin) and often showed a marked electrolyte disturbance (reflecting acid-base abnormalities) and a very mild leucocyte response; such dogs often presented as leucopenic, many being lymphocytopenic. These results indicate that the severely anaemic dogs had developed haemolytic disease (possibly immune-mediated), whereas the non-anaemic dogs had developed an acute and overwhelming inflammatory response. The mean age of the non-anaemic dogs (2.66 years) was less than the dogs in the 'severe anaemia group' (0.83 years). Dogs belonging to the traditional fighting breeds (bull terriers, pit bull terriers and Staffordshire bull terriers) were noticeably over-represented in the non-survivors of the acute inflammatory response, possibly indicating an underlying genetic basis for the different presentations. It is evident that the inflammatory-response disease presentation, which is similar to complicated falciparum malaria in humans, amy serve as an animal model for the disease.

Treatment of heartwater: potential adverse effects of furosemide administration on certain homeostatic parameters in normal sheep.

Diuretics, in particular furosemide, are generally recommended as a supportive treatment in the advanced stages of heartwater in ruminants. However, after what appeared to be possible adverse effects accompanying its use in field cases of heartwater, the effects of this drug on certain blood and urine parameters were investigated in normal sheep at the same dose rates. Diuresis with concomitant natriuresis was significant after furosemide administration, as was the expected plasma volume decrease. Other significant changes included metabolic alkalosis, hypokalaemia and reduced blood ionised calcium. The difference in duration of the diuretic effect and the duration of the changes in blood parameters from c. 3 h and c. 6 h respectively make it difficult to determine a time interval between successive treatments with furosemide. It appears that the probable cause of death of sheep with heartwater is a drastic reduction in blood volume and decreased cardiac output that leads to general circulatory failure. A therapeutic approach that involves further loss of plasma volume due to diuresis appears contradictory. The added effects of potentiating respiratory alkalosis and the terminal drop in blood ionised calcium seen in heartwater-affected animals indicate that the use of furosemide in supportive treatment of this disease is not warranted.

Morphological evidence for infection of impala, Aepyceros melampus, platelets by a rickettsia-like organism.

Ultrastructural studies revealed the presence of a parasite, believed to be Ehrlichia platys, in the blood platelets of impala. At the time of blood sampling all the animals appeared healthy. This is the first report on the presence of this rickettsia in these animals, previously described in canine platelets.

The effect of diminazene aceturate on cholinesterase activity in dogs with canine babesiosis.

A clinical trial was designed to evaluate the effects of diminazene aceturate and its stabiliser antipyrine on serum pseudocholinesterase (PChE) and red blood cell acetylcholinesterase (RBC AChE) in dogs with babesiosis. The trial was conducted on naturally occurring, uncomplicated cases of babesiosis (n = 20) that were randomly allocated to groups receiving a standard therapeutic dose of diminazene aceturate with antipyrine stabiliser (n = 10) or antipyrine alone (n = 10). Blood was drawn immediately before and every 15 minutes for 1 hour after treatment. Plasma PChE showed a 4% decrease between 0 and 60 min within the treatment group (p < 0.05). No statistically significant differences were found between the treatment and control groups at any of the time intervals for PChE. There was an increase in RBC AChE activity at 15 min in the treatment group (p < 0.05). No significant differences were found between the treatment and control groups at any time interval for RBC AChE. In view of the difference in PChE, samples from additional, new cases (n = 10) of canine babesiosis were collected to identify the affect of the drug over 12 hours. No significant depression was identified over this time interval. The results suggests that the underlying mechanism in producing side-effects, when they do occur, is unlikely to be through cholinesterase depression.

The comparative role of haemoglobinaemia and hypoxia in the development of canine babesial nephropathy.

Renal pathology associated with haemoglobinaemia resulting from Babesia canis infection is ascribed to haemoglobinuria, with or without a contribution from anaemic hypoxia. This study was undertaken to investigate the relative roles of haemoglobinaemia and hypoxia in renal function and pathology in the dog. Three groups of 6 dogs each were used over a 4-day period. The dogs in the 1st group were infused with homologous canine haemoglobin, anaemic hypoxia was induced in the 2nd group, and both treatments were applied in the 3rd group. Full urinalyses, serum urea and creatinine concentrations, fractional clearance of sodium and the activity of urine enzymes, were assessed daily. At the end of the trial period, the glomerular filtration rate (GFR) was determined and kidney specimens collected for light and electron microscopy. In the group with hypoxia only, the urine sediment contained more casts and a greater number of renal tubular epithelial (RTE) cells than in either of the other groups. Hypoxia resulted in greater enzymuria, suggestive of RTE cell pathology, whereas haemoglobinuria did not appear to have any effect on urine enzyme activity. Hypoxia resulted in a decreased GFR. Histological examination revealed a mild, single-cell tubular necrosis in the majority of the animals (all 3 groups), with granular casts in the hypoxic groups. There appeared to be a large individual variation in the ability of the kidney to handle infused haemoglobin. It was concluded that severe haemoglobinaemia did not induce a significant nephropathy, anaemic hypoxia appeared to cause a very mild nephropathy, and the combination of haemoglobinaemia and anaemic hypoxia did not exacerbate this change. These lesions were very different from those described in canine babesiosis.

Changes in haematocrit after treatment of uncomplicated canine babesiosis: a comparison between diminazene and trypan blue, and an evaluation of the influence of parasitaemia.

It has been suggested that the antibabesial drug diminazene causes a rapid decline in haematocrit after treatment of dogs with high Babesia canis parasitaemias, compared with trypan blue. To test this, 19 dogs with clinically mild to moderate, uncomplicated babesiosis were placed in low, moderate or high parasitaemia groups, based on venous parasitaemias, and were allotted randomly to diminazene or trypan blue treatment groups. Haematocrit and parasitaemia were determined before treatment, and at 2, 6, 12, 18 and 24 hours. The drugs were compared for effects on haematocrit and parasite clearance. Changes in haematocrit after treatment were analysed. There were no significant differences between diminazene and trypan blue for haematocrit or parasite clearance. There was no correlation between initial parasitaemia and initial or post-treatment haematocrit. In all dogs, haematocrit fell following treatment. The maximum mean reduction from the baseline (0 h) was 0.046 l/l (range 0.02-0.07 l/l); this most often occurred at 6 or 12 h. The 24 h haematocrit ranged from 70.5-113.6% of baseline (mean absolute haematocrit 0.019 l/l below baseline). All dogs improved clinically during the study period. It was concluded that either diminazene or trypan blue can be safely used to treat dogs with clinically mild or moderate, uncomplicated babesiosis. Parasitaemia need not be taken into account when deciding which antibabesial drug to administer and does not appear to be related to the degree of anaemia.

Met-haemoglobinuria in naturally occurring Babesia canis infection.

Higher levels of urinary met-haemoglobin were found in dogs with naturally occurring Babesia canis infection (n = 6) than in control subjects (n = 5). The urinary haemoglobin in the affected cases ranged from 1 to 4 g/l of which 28-95% was met-haemoglobin. This was a significant finding in that met-haemoglobin has been shown to be nephrotoxic in the dog, especially in the presence of aciduria. Acidic urine was also demonstrated in those dogs with B. canis infection, which may aggravate the nephrotoxic effect of the met-haemoglobinuria. Thus naturally infected B. canis cases showing haemoglobinuria may in fact have significant met-haemoglobinaemia, which may predispose them to renal disease.

Serum- and bone-mineral status of ostriches with tibiotarsal rotation.

Tibiotarsal rotation in ostrich chicks is a serious problem that accounts for considerable financial loss to ostrich farmers. Serum- and bone-mineral analyses of 20 ostrich chicks with tibiotarsal rotation were compared with serum- and bone-mineral analyses of eight normal ostrich chicks of comparable age, sex and body mass, and raised under identical conditions. The serum-zinc values were significantly higher and the bone-calcium and phosphorus values significantly lower in the affected group than in the group of normal ostrich chicks. The results indicated poor mineralization of bone with subsequent reactive osteoid formation.

The clinical pathology of heartwater. III. Changes in blood clotting, blood calcium, blood protein, haematocrit and white-cell counts in sheep with experimentally induced heartwater.

Studies to evaluate changes in blood clotting, blood calcium and protein, the haematocrit and white-cell counts were undertaken in seven sheep with experimentally induced heartwater. A marked decline in thrombocyte count was recorded during the acute stage of the disease. This was associated with increases in both prothrombin time (PT) and activated partial thromboplastin time (APTT); fibrinogen increased while there was no detectable increase in fibrinogen degradation products (FDP). At the same time total serum protein (TSP), albumin and globulin dropped very sharply; total calcium showed a progressive drop but ionized calcium rose initially and was followed by a terminal decline. The total leucocyte count showed a terminal increase while the haematocrit dropped progressively.

Changes in the blood-gas status of sheep with experimentally induced heartwater.

The blood-gas status of seven sheep with experimentally induced heartwater during the acute and terminal stages was investigated. Changes in blood gas included a decline in arterial oxygen tension (pO2) combined with a respiratory alkalosis. Although the sheep became hypoxaemic, blood-gas changes associated with respiratory failure were not observed.

A comparison between the disease status of hospitalized dogs from developed and those from developing communities.

The health status of canine populations within developed and those within developing communities was studied in a retrospective survey and compared. There were significant differences in the prevalence of disease amongst the hospitalised dogs from the 2 communities. Dogs from developing communities were mainly young cross-bred dogs which suffered from infectious diseases (44%), trauma (22%) and parasitic diseases (11%). There was a high mortality rate (30%) and 82% of these patients suffered from diseases that could have been prevented. The dogs from developed communities were mainly adult or old pure-bred dogs that suffered mainly from organ diseases (57%). There was a low mortality rate (10%) while only 31% suffered from diseases that could have been prevented. Based on the epidemiological findings, it was evident that owners of dogs in the developing communities required education in primary and secondary prevention of disease.

Susceptibility of dogs to West Nile virus: a survey and pathogenicity trial.

A serological survey of dogs from the highveld region of South Africa showed that 37 per cent (138 of 377) had neutralizing antibodies to West Nile (WN) virus and only 2.7 per cent (10 of 377) had antibodies to Sindbis virus. WN virus was isolated from one of the WN-antibody negative sera. Because these results suggested that dogs may play an important part in the epidemiology of WN virus, a pathogenicity trial was carried out. Two of three dogs infected with WN virus had a mild recurrent myopathy, but no other abnormalities were detected in the biochemical or haematological tests performed on any of the dogs. All three dogs developed antibodies but a low titre-viraemia was detected in only one dog. It was concluded that dogs do not play an important part in the epidemiology of WN virus but they may play a small part in the maintenance of the virus.