Upregulation of sialyltransferases ST3Gal1 and ST6Gal1 promotes stabilization of erythrocyte mass and recovery of anemia in Trypanosoma brucei brucei-infected pigs.

The role of Sialyltransferases (STs) specifically subfamilies ST3Gal1 and ST6Gal1 tissue expression was investigated in the liver and kidney of Trypanosoma brucei brucei-infected and uninfected control pigs. The study was aimed to provide emerging target for treatment. Pigs were experimentally infected with 2 × 106 T. b. brucei (Federe strain); parasitemia was monitored by microscopy and tissue expression levels of ST3Gall and ST6Gall in the liver and kidney were assessed using quantitative real-time polymerase chain reaction (qRT-PCR). Parasitemia were undulating and anemia occurred significantly (P < 0.01) on day 13 in the infected pigs with an attempt to recover toward the termination of the study on day 21. The gene expressions for hepatic and renal ST3Gal1 and ST6Gal1 were significantly (P < 0.0001) upregulated 5-42 folds in the infected pig compared to the non-infected control group. It was concluded from the findings in this study that increased tissue expression of ST3Gal1 and ST6Gal1 in T b. brucei-infected pigs may play a pivotal role in the resialylation of desialylated red blood cells, thereby promoting recovery of the red blood cells and stabilization of erythrocyte mass in trypanosome-infected pigs. It is recommended that the expression of serum ST3Gal1 and ST6Gal1 be investigated further, in trypano-susceptible against trypano-tolerant breeds of animals to determine the role of these genes in trypano-tolerance.

Anemia amelioration by lactose infusion during trypanosomosis could be associated with erythrocytes membrane de-galactosylation.

African trypanosomosis is a potentially fatal disease that is caused by extracellular parasitic protists known as African trypanosomes. These parasites inhabit the blood stream of their mammalian hosts and produce a number of pathological features, amongst which is anemia. Etiology of the anemia has been partly attributed to an autoimmunity-like mediated erythrophagocytosis of de-sialylated red blood cells (dsRBCs) by macrophages. Lactose infusion to infected animals has proven effective at delaying progression of the anemia. However, the mechanism of this anemia prevention is yet to be well characterized. Here, the hypothesis of a likely induced further modification of the dsRBCs was investigated. RBC membrane galactose (RBC m-GAL) and packed cell volume (PCV) were measured during the course of experimental trypanosomosis in mice infected with Trypanosoma congolense (stb 212). Intriguingly, while the membrane galactose on the RBCs of infected and lactose-treated mice (group D) decreased as a function of parasitemia, that of the lactose-untreated infected group (group C) remained relatively constant, as was recorded for the uninfected lactose-treated control (group B) animals. At the peak of infection, the respective cumulative percent decrease in PCV and membrane galactose were 30 and 185 for group D, and 84 and 13 for group C. From this observed inverse relationship between RBCs membrane galactose and PCV, it is logical to rationalize that the delay of anemia progression during trypanosomosis produced by lactose might have resulted from an induction of galactose depletion from dsRBCs, thereby preventing their recognition by the macrophages.

Survey of bovine trypanosomosis in the Kachia Grazing Reserve, Kaduna State, Nigeria.

This study assessed the prevalence of trypanosomes in cattle at the Kachia Grazing Reserve (KGR) in March and June 2004 and in February 2005. A total of 1293 cattle blood samples were collected at random. The samples were analysed using the buffy coat technique and Giemsa thin blood films for parasite detection and identification. The effects of herd pen location to watering and grazing point's distances (using the global positioning system (GPS)) were determined and mean Packed cell volume (PCV) assessed. Overall, the detected prevalence of trypanosomosis was 8.4%, much higher than the previous prevalence of 5.3% before the present study was conducted. The prevalences in the months of March, June (2004) and February (2005) were 2.3%, 11.6% and 15.4%, respectively. Increased prevalence was associated with proximity of herd pens to watering point's distances (chi(2) for linear trend=4.447, P<0.05), but no association of herd pens to grazing point distances (chi(2)=2.186, P>0.05); suggesting that hydrological network played an important part in trypanosomosis transmission. The mean PCV of parasitaemic and apparasitaemic cattle were respectively 25.99+/-1.82% and 29.31+/-1.70%. The drop in mean PCV was most in 0-1-year age group, 23.47+/-3.10% and was statistically significant (P<0.05), suggesting that anaemia was most pronounced in this age group. Factors that may have contributed to the increased prevalence obtained were collapse of control measures and breed susceptibility. Since, Zebu cattle were the predominant breeds in the reserve, the study advocates effective use of insecticide impregnated screens (traps and targets) with community participation in mind for sustainability. If government intervenes through PATTEC ground spraying of insecticides in the reserve is recommended. In addition, chemotherapeutic and chemoprophylaxis should be systematically used to fight the problem of trypanosomosis in the KGR towards improved livestock production.

Studies on effects of lactose on experimental Trypanosoma vivax infection in Zebu cattle. 2. Packed cell volume.

The ability of intravenously administered lactose in normal saline to prevent a decline in packed cell volume (PCV) during experimental trypanosomosis was studied in Zebu cattle. During the lactose infusion period, the PCV was stable up to Day 5 post-infection (p.i.) in a lactose-infused group, compared to that in an uninfused group in which the PCV dropped significantly (P < 0.05) as shown by the values of cumulative percentage change. Furthermore the mean rate of change in PCV was significantly (P < 0.05) higher in the uninfused group relative to the lactose-infused group during the same period. While the PCV fell markedly in the lactose-infused group a day after lactose infusion was stopped (Day 13 p.i.), subsequent PCV values were significantly (P < 0.05) higher compared to those in the uninfused group, up to the end of experiment on Day 17 p.i. However the mean rates of change in PCV did not vary significantly (P > 0.05) between the groups during the period in which lactose infusion was stopped. The mean levels of parasitaemic waves and parasitaemia were higher, more prolonged and more frequent in the lactose-infused group. It was inferred that the lactose was able to prevent an early onset of anaemia in the Trypanosoma vivax-infected Zebu cattle.

Studies on effects of lactose on experimental Trypanosoma vivax infection in Zebu cattle. 1. Plasma kinetics of intravenously administered lactose at onset of infection and pathology.

Lactose in normal saline was administered intravenously to a group of Zebu cattle infected with Trypanosoma vivax to determine the blood plasma kinetics at onset of an experimental infection and its ability to protect tissues against damage as part of preliminary studies to determine its suitability for use in the treatment of trypanosomosis. Significantly (P < 0.01) higher lactose concentrations were observed in the T. vivax-infected bulls at 30 min and 1 h (P < 0.05) post-infection (p.i.) and by 4 h p.i. the plasma lactose remained above the level prior to infusion, after which it fell slightly below the pre-infusion level in the uninfected group. Calculated pharmacokinetic parameters revealed delayed excretion of lactose in the T. vivax-infected group soon after infection. The total body clearance (Cl(B)) was significantly (P < 0.05) reduced. The biological half-life (t1/2), elimination rate constant (k(el)) and apparent volume of distribution (V(d)) were relatively decreased (P > 0.05) as a result of the T. vivax infection. Retention of lactose in the plasma was attributed to decreased plasma clearance. It is suggested that the presence of trypanosomes in circulation rather than organic lesions could have been responsible for the delay observed in the excretion of lactose. At 12 weeks p.i., when the experiment was terminated, the group infected and given lactose infusion (despite higher parasitaemia) had no gross or histopathological lesions in the brain, spleen, lymph nodes, heart, kidneys, liver and testes. However, the group infected but not infused with lactose were emaciated, had pale mucosae, watery blood, general muscular atrophy, serous atrophy of coronary fat and other adipose tissue, hepatomegaly, splenomegaly, swollen and oedematous lymph nodes, all of which are suggestive of trypanosomosis. Histopathological lesions included narrowing of Bowman's space and hypercellularity of glomerular tufts in the kidneys with the mean glomerular tuft nuclear indices (GTNs) in the group significantly higher (P < 0.01) than the mean GTNs of the lactose-infused and control bulls. Degenerative changes occurred in the myocardium, spleen, testes and epididymides. The tesicular and epididymal lesions are indicative of male reproductive dysfunction.

Toxicological screening of lyophilized extract of some Nigerian wild mushrooms in mice.

Mushrooms are macrofungi widely consumed as food. However, many mushrooms rot away in the wild because of fear of toxicity. Therefore, lyophilized aqueous extracts of 6 mushroom species collected from Zaria, Nigeria and taxonomically identified as Chlorophyllum molybdites, Panaeolus subalteatus, Macrolepiota procera, Leucopaxillus albissmus, Hygrophoropsis aurantiacus and Pholiota aurea were screened for toxicity in mice. Lyophilized aqueous extract of each of these mushrooms was administered to three groups of 3 mice intraperitoneally (i.p.) at doses of 100, 1000 and 10, 000 mg kg(-1), respectively. Another group of three mice given distilled water served as control. The mice were examined for clinical signs of toxicity over a period of 72 h and pathological examinations conducted on dead animals. The severity of clinical signs, onset of death and pathological lesions were dose dependent. Death occurred within 10 min in all the mice dosed at 10,000 mg kg(-1) with the lyophilized extracts of all the mushrooms screened, with the exception of that of H. aurantiacus, which produced death 21-23 h post administration. This result showed that all the screened mushrooms, including the popular edible M. procera were found toxic. Therefore, since all the mushrooms screened were found toxic, it is recommended that extreme caution should be exercised in their consumption. Furthermore, in view of the regional differences in the toxicity of mushrooms, there is the need to screen more wild mushrooms found in Nigeria for toxicity. This will boost mushroom mycophagy, reduce poisoning incidence and reduce wastage of edible mushrooms in the wild.

Sequential testicular and epididymal damage in Zebu bulls experimentally infected with Trypanosoma vivax.

Six Zebu bulls aged between 31 and 34 months exhibiting good libido were used to study sequential testicular and epididymal damage in Trypanosoma vivax infection. Three bulls were infected with T. vivax, while the other three served as controls. All infected bulls became parasitaemic by day 5 post-infection and developed clinical trypanosomosis with rapidly developing anaemia. Representative bulls, one from each of the infected and control groups, were sacrificed on days 14, 28 and 56 post-infection. Testes and epididymides from these animals were studied histopathologically after processing and staining with haematoxylin and eosin (H and E). Testicular degeneration developed in all the infected bulls characterized by depletion of spermatogenic cells and destruction of interstitial tissue. The most severe testicular degeneration occurred in the bull that was sacrificed 56 days post-infection. Epididymal sperm reserves were 36%, 4% and 0%, respectively, in infected bulls that were sacrificed on days 14, 28 and 56 post-infection. The 0% epididymal sperm reserve may suggest complete cessation of spermatogenesis. It was concluded from this study that T. vivax infection of Zebu bulls could cause severe testicular and epididymal damage that may result in infertility or even sterility of the affected animals at early infection stages not previously thought.

Trace minerals in serum of sheep infected with Trypanosoma congolense.

Yankassa sheep (20) were grouped into A and B and infected with Trypanosoma congolense isolated from a cow and maintained in mice. Two milliliter x 10(7) parasites were used to infect group A. The course of the infection and serum trace minerals (Iron, (Fe) and Copper, (Cu) were studied and determined using Atomic Absorption Spectrophotometer (AAS). There was significant drop in concentration of iron (p<0.001) Post Infection (pi) while that of copper, no significant change (p>0.05). The values of the contemporaneously uninfected control sheep were significantly higher for iron and not for copper. Sheep are susceptible to isolate from cow and passaged in mice and with the fluctuating concentrations of Fe and consistency of Cu, it may suggest that these minerals may have a role in the pathogenesis of trypanosomosis due to T. congolense.

Characterization of a sialidase (neuraminidase) isolated from Clostridium chauvoei (Jakari strain).

A sialidase from Clostridium chauvoei (Jakari strain), an indigenous bacterial strain that causes blackleg in Nigerian cattle and other ruminants was isolated and partially purified by chromatography on DEAE cellulose, hydroxyapatite and phenyl agarose columns. The enzyme migrated as a 65-kDa protein after electrophoresis on sodium dodecyl sulphate polyacrylamide gels. It was optimally active at pH 4.5 and 40 degrees C with an activation energy (Ea) of 13.40 kJ mol(-1). It had Km and Vmax values of 170 microM and 200 micromole h(-1) mg(-1) respectively with fetuin as substrate. When sialyllactose (Neu5Ac2,3 lactose) was used as substrate the Km and Vmax values were 8 microM and 5 micromoles min(-1) mg(-1) respectively. The Clostridium chauvoei sialidase cleaved sialic acids from RBC ghosts of sheep, horse, goat, cattle, pig and mice as well as mouse brain cells, albeit at different rates. The enzyme was activated by Ca2+ and Mg2+ and inhibited by the group-specific reagents diethylpyrocarbonate (DEP) and N-ethylmalemide (NEM). The sialidase inhibitors, 2,3 didehydroneuraminic acid (Neu5Ac2,3en) and paranitrophenyl oxamic acid (pNPO) inhibited the enzyme competitively with Ki values of 40 and 30 microM respectively.

Comparative studies on erythrocyte sialic acid levels in apparently healthy indigenous Nigerian poultry species.

The erythrocyte surface sialic acid levels of 200 apparently healthy indigenous Nigerian poultry species (pigeons, guinea fowls, ducks and chickens, n = 50 for each species) presented for slaughter at a poultry abattoir in Zaria, Nigeria was determined. Other parameters determined were packed cell volume (PCV) and plasma total protein (TP) concentration. The mean erythrocyte surface sialic acid concentration of pigeons, guinea fowls, ducks and chickens were 7.88 +/- 2.51, 14.6 +/- 2.51, 17.6 +/- 2.51 and 14.2 +/- 2.51 mg mg(-1) respectively. There was a statistically significant difference between the mean erythrocyte surface sialic acid concentration of all the species of poultry sampled (P < 0.05). The high erythrocyte surface sialic acid concentration in the indigenous Nigerian poultry species could be responsible for their resistance to infectious diseases, whose aetiologic agents produce neuraminidases. The mean PCV of the pigeons, guinea fowls, ducks and chickens were 46.22 +/- 6.91, 38.24 +/- 6.91, 36.50 +/- 6.91 and 36.46 +/- 6.91% respectively. The difference between the mean PCV values of pigeons on the one hand and guinea fowls, ducks and chickens on the other was statistically significant (P < 0.05). A negative correlation (r = -0.36, P < 0.05) between mean erythrocyte surface sialic acid concentrations and PCV was observed, i.e. the birds with the highest mean PCV values had the lowest levels of erythrocyte surface sialic acid. There was no correlation between TP concentration and either erythrocyte surface sialic acid concentration or PCV values. It is suggested, based on this study, that erythrocyte sialic acid types in these species should be determined, as the results may be vital in selective breeding.

Characterization of sialidase from bloodstream forms of Trypanosoma vivax.

Sialidase (EC: 3.2.1.18) from Trypanosoma vivax (Agari Strain) was isolated from bloodstream forms of the parasite and purified to apparent electrophoretic homogeneity. The enzyme was purified 77-fold with a yield of 32% and co-eluted as a 66-kDa protein from a Sephadex G 110 column. The T. vivax sialidase was optimally active at 37 degrees C with an activation energy (E(a)) of 26.2 kJ mole(-1). The pH activity profile was broad with optimal activity at 6.5. The enzyme was activated by dithiothreitol and strongly inhibited by para-hydroxy mercuricbenzoate thus implicating a sulfhydryl group as a possible active site residue of the enzyme. Theenzyme hydrolysed Neu5Ac2,3lac and fetuin. It was inactive towards Neu5Ac2,6lac, colomic acid and the gangliosides GM1, and GDI. Initial velocity studies, for the determination of kinetic constants with fetuin as substrate gave a V(max) of 142.86 micromol h(-1) mg(-1) and a K(M) of 0.45 mM. The K(M) and V(max) with Neu5Ac-2,3lac were 0.17 mM and 840 micromole h(-1) mg(-1) respectively. The T. vivax sialidase was inhibited competitively by both 2,3 dideoxy neuraminic acid (Neu5Ac2,3en) and para-hydroxy oxamic acid. When ghost RBCs were used as substrates, the enzyme desialylated the RBCs from camel, goat, and zebu bull. The RBCs from dog, mouse and ndama bull were resistant to hydrolysis.

Haemagglutination inhibition antibodies, rectal temperature and total protein of chickens infected with a local Nigerian isolate of velogenic Newcastle disease virus.

Changes in values of haemagglutination inhibition (HI) antibody titre, rectal temperature (RT) and total protein (TP) were determined for Shaver Brown chickens infected with Newcastle disease virus (NDV) Kudu 113. The infected chickens came down with Newcastle disease by day 3 post infection (PI). The major clinical signs were depression, greenish diarrhoea, paralysis of legs and wings, opisthotonus and torticolis. Mortality and morbidity were 52% and 1000%, respectively. There were haemorrhagic lesions in the wall of the intestine, proventricular mucosa and caecal tonsils. There were necrosis and mononuclear cell infiltration of the liver, kidney and spleen. There was a significant increase in daily mean HI antibody titres from days 3 to 9 PI. Similarly, significant rise in daily mean RTs were noticed in the infected chickens from days 1 to 13 PI. On the other hand, there was a decrease in daily mean TP concentrations of infected chickens, beginning from day 3 PI, and the lowest concentration of 2.60 +/- 0.15 g/dl was obtained by days 7 and 11 PI. The values of HI, RT and TP for the control chickens were relatively constant during the experiment. The correlation coefficient (r) between HI and RT was positive and highly significant (r = 0.725, p<0.001), while the relationship between HI and TP was negative but highly significant (r = -0.712, p<0.001). It was concluded that NDV Kudu 113 induced increases in values of HI and RT, which occurred concurrently with a decrease in TP concentrations of infected chickens.

The inhibition of Clostridium chauvoei (jakari strain) neuraminidase activity by methanolic extracts of the stem barks of Tamarindus indicus and Combretum fragrans.

The inhibition of neuraminidase from Clostridium chauvoei (jakari strain) with partially purified methanolic extracts of some plants used in Ethnopharmacological practice was evaluated. Extracts of two medicinal plants, Tamarindus indicus and Combretum fragrans at 100-1000 microg/ml, both significantly reduced the activity of the enzyme in a dose-dependent fashion (P < 0.001). The estimated IC50 values for Tamarindus indicus and Combretum fragrans were 100 and 150 microg/ml respectively. Initial velocity studies conducted, using fetuin as substrate revealed a non-competitive inhibition with the Vmax significantly altered from 500 micromole min(-1) mg(-1) to 240 micromole min(-1) mg(-1) and 340 micromole min(-1) mg(-1) in the presence of Tamarindus indicus and Combretum fragrans respectively. The KM remained unchanged at 0.42 mM. The computed Index of physiological efficiency was reduced from 1.19min(-1) to 0.57min(-1) and 0.75min(-1) with Tamarindus indicus and Combretum fragrans as inhibitors respectively.

Aetiology and prevalence of canine anaemia in Zaria: a review of 2139 cases observed at the Veterinary Teaching Hospital of the Ahmadu Bello University, Zaria, Nigeria (1990-2003).

An investigation was conducted at the Ahmadu Bello University Veterinary Teaching Hospital (ABUVTH) between January, 1990 and September, 2003 to determine the aetiology and prevalence of canine anaemia in Zaria, Nigeria. Out of the 5278 mongrel dogs presented during the period 1990-2003, 2139 (40.5%) were found to be anaemic, with packed cell volume (PCV) values ranging from 7 to 36%. The clinical signs presented by these dogs include: Pale mucous membranes, weakness, depression, anorexia/inapettence and reduced activity. About 50 dogs (about 1%) with helminths and haemoparasitic infestations had high PCV values (37-40%) without clinical presentation of anaemia or disease. Most of the dogs with anaemia (n = 2016 or 94.2%) had parasitic infestations. About 1580 (about 74%) of the anaemic cases, attributed to parasitic infestations occurred between May and October. A few dogs (n = 55, 2.6%) had anaemia due to poor nutrition, while 68 (3.2%) had anaemia with unknown cause. The public health significance of the parasites reported in this study is discussed.

Pathogenesis and pathology of blackleg in ruminants: the role of toxins and neuraminidase. A short review.

There is no consensus on the pathogenesis of blackleg infection that occurs in ruminants, but toxins and neuraminidase produced by Clostridium chauvoei are believed to play a significant role in the pathogenesis of the disease. This paper provides an update on the role of toxins and neuraminidase in the pathogenesis and pathology of the disease. The use of neuraminidase inhibitors to manage clinical blackleg infections is therefore an alternative therapeutic protocol that should be thoroughly investigated. It is suggested that in vivo clinical trials should be carried out to determine the mechanism of action and clinical efficacy of neuraminidase inhibitors.