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A J Trees

Publications and source records attributed to A J Trees.

At least 73 records · Page 4Linked to original sources

Clinical aspects of 27 cases of neosporosis in dogs.

Twenty-seven cases of neosporosis in European dogs are described. The disease was confirmed by immunohistochemistry, electron microscopy, or a favourable response to treatment in the dogs with appropriate clinical signs, and by the presence of antibodies to Neospora caninum but not to Toxoplasma gondii. The affected dogs were two days to seven years old, and of 13 different breeds. Both sexes were affected and in most cases littermates remained normal. Twenty-one cases had an initial hindlimb paresis or ataxia, in which muscle atrophy was the most consistent clinical sign. Rigid hyperextension developed in approximately half of the cases. Anorexia and pyrexia were rare. Other clinical signs included forelimb ataxia, head tremors with tetraparesis and sudden collapse due to myocarditis. Titres of > or = 1:800 in the N caninum indirect fluorescent antibody test were detected in the 20 cases from which serum samples were taken. Such high titres are rare in healthy dogs and strongly suggest a diagnosis of neosporosis. Sixteen of the dogs received appropriate antiprotozoal treatment with clindamycin, potentiated sulphonamides and/or pyrimethamine; 10 made a full or functional recovery. Recovery was less likely in peracute cases with severe clinical signs, and when the treatment was delayed.

Animals↗

Use of excretory/secretory antigens for the serodiagnosis of Anoplocephala perfoliata cestodosis.

Whole worm extract (WWE) and excretory/secretory (E/S) antigens of Anoplocephala perfoliata were characterised by SDS-PAGE and their use in the serodiagnosis of equine cestodosis was evaluated. An enzyme-linked immunosorbent assay (ELISA) was used to compare WWE and E/S antigen as the capture layer in an antibody capture ELISA. E/S antigen gave the best differentiation between sera from tapeworm-positive and tapeworm-negative horses. The E/S-ELISA was optimised and validated against sera from horses of known tapeworm status. This assay gave a diagnostic sensitivity of 68% (n = 38) and a specificity in helminth-naive horses of 95% (n = 20). Western blot analysis of tapeworm-positive and tapeworm-negative sera revealed the immunoreactive proteins of E/S antigen to be those of 12 and 13 kDa.

Animals↗

In vitro responsiveness of Babesia bovis to imidocarb dipropionate and the selection of a drug-adapted line.

A South African stock of Babesia bovis was successfully resuscitated from liquid nitrogen, and cultured in microaerophilous stationary phase. The in vitro susceptibility of the B. bovis stock to titrated concentrations of imidocarb dipropionate was observed and the 50% inhibitory concentration (IC) was determined (8.7 x 10(7) g ml-1). A drug-adapted line was developed by culture in the presence of sub-inhibitory concentrations of imidocarb dipropionate and it had an IC50 eight times higher than that of its original stock (6.6 x 10(-6) g ml-1). The drug-adapted line was cryopreserved and resuscitated from liquid nitrogen. Continuous culture of the non-drug adapted line through 15 subcultures did not change the IC50 (8.3 x 10(-7) g ml-1).

Animals↗

Susceptibility of Brugia malayi and Onchocerca lienalis microfilariae to nitric oxide and hydrogen peroxide in cell-free culture and from IFN gamma-activated macrophages.

The susceptibility of Brugia malayi and Onchocerca lienalis microfilariae to H2O2 and NO either in cell-free culture or from IFN gamma-activated macrophages was examined. In cell-free culture, O. lienalis microfilariae were highly susceptible to H2O2 induced toxicity, exhibiting rapid reductions in motility and viability. The addition of exogenous catalase abrogated H2O2-induced killing. In contrast, B. malayi microfilariae were relatively resistant to H2O2, with concentrations as high as 50 microM having no effect on motility or viability. On exposure to NO, both species showed reductions in motility within 5-30 min, but longer was required to see effects on the viability of microfilariae. Parasites incubated with IFN gamma-activated macrophages also exhibited marked reductions in motility and viability. In cultures with B. malayi and activated macrophages, inhibition of these effects was achieved by the addition of either L-NMMA, to abolish NO production, or neutralizing anti-TNF alpha antibodies. Attempts to inhibit parasite killing by the addition of catalase to macrophage cultures were ineffective. The results of this study show that B. malayi and O. lienalis microfilariae have different susceptibility to H2O2, but are equally affected by exposure to NO. Moreover both species are killed by IFN gamma-activated macrophages and in the case of B. malayi, killing is dependent on the generation of NO via TNF alpha.

Animals↗

Correlation of antigen specific IgG and IgG(T) responses with Anoplocephala perfoliata infection intensity in the horse.

There is increasing interest in the application of serological methods to macro-parasite infections to indicate infection intensity, which in turn is related to pathogenicity. Colic is the single most important cause of mortality in horses and there is evidence that a proportion of colic cases are associated with infection with the intestinal cestode Anoplocephala perfoliata. In order to develop better tools to investigate this association, the correlation between antigen-specific equine IgG and IgG(T) and infection intensity of A. perfoliata was investigated. Affinity purification of a 12/13 kDa protein doublet from crude excretory/secretory (E/S) products, and its use in enzyme linked immunosorbent assays (ELISA) is described. Its use in the immunodiagnosis of equine cestodosis and the correlation of anti-12/13 kDa IgG and IgG(T) with parasite burden is investigated using sera from 94 horses of known tapeworm infection intensity. The anti-12/ 13 kDa IgG and IgG(T) ELISAs gave correlation coefficients with infection intensity of 0.56 and 0.63 respectively. Linear regression analysis also indicated that anti-12/ 13 kDa IgG(T) was the best predictor of infection intensity. The decay of anti-12/13 kDa IgG(T) in horses following the elimination of A. perfoliata is demonstrated for four horses. Specificity of the anti-12/13 kDa IgG(T) ELISA is investigated with sera from 33 A. perfoliata negative horses with other helminth infections. Immunoblotting studies demonstrate no cross-reactivity between A. perfoliata 12/13 kDa antigen and the protein antigens of other helminths. It is concluded that assay of anti-12/13 kDa IgG(T) provides a useful tool for the assessment of A perfoliata infection intensity for clinical diagnosis and for epidemiological studies.

Animals↗

Distribution of Neospora caninum within the central nervous system and other tissues of six dogs with clinical neosporosis.

Six cases of neosporosis in dogs were diagnosed on the basis of clinical signs, serology and immunohistochemistry. The brains and spinal cords, at several levels, and various other tissues were examined using anti-Neospora caninum serum by an immunoperoxidase technique. Parasite density varied markedly from case to case. Although found most consistently in the cerebrum, parasites were distributed throughout the central nervous system (CNS), both within the grey and white matter and within nerve roots. Clinical signs were not related to the position of parasites. Tissue cysts were found infrequently in all areas of the CNS, but not in other tissues. Parasite density was not related to the age of the dog or whether treatment had been given. However, with the exception of the only adult dog examined, more CNS parasites were found in dogs with a longer duration of illness and with higher antibody titres. Tachyzoites were present in skeletal muscles, in the muscularis of the oesophagus, in heart, lung and, less frequently, liver, and rarely in the adrenal gland, thyroid gland and uterus; no clinical signs were seen resulting from damage to these organs. Parasites were not observed in lymphoid tissue. In visceral organs, parasites were most widely distributed in peracute cases. For post mortem diagnosis of neosporosis, the CNS, particularly the cerebrum, is the optimum tissue to examine but parasites may also be found in many other tissues, especially in acute cases. Muscle biopsy of appropriate muscles (as suggested by the clinical signs) provides the possibility of a definitive premortem diagnosis.

Animals↗

Characterization and immunoprotective properties of a monoclonal antibody against the major oocyst wall protein of Eimeria tenella.

The oocyst wall of Eimeria spp. consists of a 10-nm-thick outer lipid layer and a 90-mm-thick inner layer of glycoprotein which has been described previously to be composed of a single major protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions and (125)I labelling of a oocyst wall fragments and of delipidated intact oocysts revealed a molecule of approximately 12 kDa as the major protein component of the oocyst wall of Eimeria tenella. An immunoglobulin M monoclonal antibody (c11B9F3) was produced against this 12-kDa oocyst wall protein sliced from a preparative SDS-polyacrylamide gel. Its reactivity by immunofluorescence against oocyst wall fragments and sporozoites or by immunoperoxidase assays of infected tissue sections was stage restricted to gametocytes and oocysts but pan-specific against all face of the oocyst wall. In chicks passively immunized with C11B9F3, oocyst output was significantly (P<0.01) reduced by 42 to 54% after homologous E. tenella infection and by 35% after heterologous Eimeria maxima infection compared with that of control groups. The results demonstrate the presence of a highly conserved, low-molecular-weight antigen on the oocyst wall and the gametocytes of Eimeria spp. which is a candidate for inclusion in a pan-specific, transmission-blocking vaccine against avian coccidiosis.

Animals↗

Experimental neosporosis in pregnant ewes and their offspring.

Six groups of six pregnant ewes each were inoculated with 170,000 or 1,700,000 tachyzoites of Neospora caninum on gestation day 65, 90, or 120. All ewes seroconverted, and none showed signs of illness other than abortion. Regardless of the inoculum dose, all ewes inoculated on gestation day 65 aborted; ewes inoculated on gestation day 90 aborted, gave birth to weak lambs, or gave birth to clinically normal lambs; and all ewes inoculated on gestation day 120 gave birth to clinically normal lambs. Using an immunohistological procedure that stains bradyzoites, we observed protozoal cysts in brains of 11 of 29 (38%) aborted fetuses, in one of four (25%) weak lambs, and in seven of 18 (39%) clinically normal lambs. Cysts were not observed in extraneural tissues from two clinically normal lambs that had cysts in the brain. No evidence of infection was observed in tissues of five ewes examined using an immunohistological procedure that stains N. caninum tachyzoites and bradyzoites. Multifocal nonsuppurative encephalitis was observed in 46 of 51 (90%) aborted, weak, or clinically normal lambs. Cerebral necrosis, dystrophic mineralization, and meningitis were also commonly identified in live and aborted lambs (even when severely autolyzed). Nonsuppurative, necrotizing placentitis was observed in 15 of 17 (88%) placentas. Nonsuppurative myositis was common in fetuses but not in live lambs. Inflammation occurred less frequently in liver and lung. Clinical and pathological features of neosporosis in sheep closely resemble those of bovine neosporosis and ovine toxoplasmosis. Although abortion caused by naturally occurring neosporosis in sheep has not been reported, diagnosticians should carefully distinguish between neosporosis and toxoplasmosis in cases of ovine protozoal abortion unless future investigations exclude the likelihood of naturally acquired neosporosis in sheep.

Abortion, Veterinary↗

Effects of haemagglutination (lectin) inhibitory sugars on Theileria parva infection in Rhipicephalus appendiculatus.

We have previously described the presence of haemagglutinins in tissues of the tick, Rhipicephalus appendiculatus and determined their sugar specificities by inhibition experiments. In this study, haemagglutination inhibitory sugars are shown to have an effect in vivo on the abundance of Theileria parva infected salivary gland acini in Rhipicephalus appendiculatus. A significant increase (P < 0.05) was observed in T. parva acinar infection rates in the salivary glands of R. appendiculatus fed on ears of rabbits infused with melibiose and raffinose. In contrast, mannose and turanose (non-haemagglutination inhibitory sugars) did not cause elevation of T. parva acinar infection rates. The effect of melibiose in elevating acinar infections was observed when used only during T. parva maturation in the salivary glands but not during parasite pick-up from an infected bovine host. Stabilates produced from ticks with elevated acinar infections did not differ from control stabilates in infectivity to cattle, by comparison of prepatent periods to pyrexia, or parasitosis, or in the severity of reactions.

Animals↗

Interferon gamma inhibits the intracellular multiplication of Neospora caninum, as shown by incorporation of 3H uracil.

An assay was developed to quantify the growth of two different isolates of the protozoon Neospora caninum within ovine fibroblast cells in vitro by differential uptake of 3H uracil. The NC-1 isolate of N. caninum multiplied more quickly in culture than the NC Liverpool isolate, as reflected by increased incorporation of isotope by the former over a shorter period of time. After the parasites had left the ruptured host cells, there was very little incorporation of isotope. This suggested that multiplication occurred within and not outside the cells. Treatment of the cells with ovine recombinant interferon gamma for 24 h before infection significantly inhibited intracellular multiplication of the parasite.

Animals↗

Characterization of the first European isolate of Neospora caninum (Dubey, Carpenter, Speer, Topper and Uggla).

Neospora caninum is an apicomplexan, protozoan parasite, which causes severe disease in dogs and cattle. It has previously been isolated only in the United States. A 5-week-old Boxer pup with a progressive hindlimb paresis was diagnosed as suffering from neosporosis on the basis of clinical signs and the presence of anti-Neospora antibodies in it, 2 litter-mates and its dam. Despite treatment with sulphonamides, the pup was euthanased 3 days later. The diagnosis of neosporosis was confirmed by immunohistochemical examination of muscle and CNS tissue sections from the pup. Parasites were isolated into Vero cell culture from the cerebrum, and confirmed as Neospora caninum by immunofluorescence with specific antibody, tachyzoite ultrastructure and 16S-like ribosomal RNA sequences. This isolate (designated NC-Liverpool) has been continuously passaged every 7-10 days. Its growth characteristics, ultrastructure and antigenic profile, as revealed by immunoblotting, have revealed no major differences from the American NC-1 isolate. Furthermore, no difference was seen when comparing the sequences of 16S-like ribosomal RNA and the ITS1 region of the two isolates.

Animals↗

Cellular and humoral immune responses to recombinant antigens in sheep infected with Toxoplasma gondii.

Immune responses to recombinant fragments of the Toxoplasma gondii antigens ROP2 and GRA2 were investigated in sheep naturally and experimentally infected with T. gondii oocysts. Specific serum antibodies to C-terminal fragments were detected by ELISA. Cell-mediated responses in peripheral blood mononuclear cells were demonstrated by proliferation and interferon-gamma production following in vitro stimulation with the ROP2 fragment. This data indicates the presence of epitopes for sheep B cells in the recombinant GRA2 fragment and for both B and T cells in the ROP2 fragment.

Animals↗

Evaluation of suramin, ivermectin and CGP 20376 in a new macrofilaricidal drug screen, Onchocerca ochengi in African cattle.

To aid the development of a macrofilaricidal agent for Onchocerca volvulus, the African bovine parasite, O. ochengi, was evaluated as a drug screen by testing three known filaricidal drugs. Groups of five Zebu cattle, naturally infected with more than 15 palpable O. ochengi nodules in the ventral skin, were treated with either suramin (10 mg/kg/day i.v. for 6 days), ivermectin (200 micrograms/kg, s.c.), CGP 20376 (20 mg/kg orally) or left untreated as controls and examined at intervals up to 137 days post-treatment (d.p.t.). After ivermectin treatment, microfilarial densities in the skin decreased within one week to virtually zero and remained at a very low level. A similar rapid and profound reduction was seen after CGP 20376 treatment, but by 137 d.p.t. microfilarial skin densities were approaching pre-treatment levels. With suramin, skin microfilarial densities fell to very low levels after 12 weeks but rose slightly by 137 d.p.t. Effects on the macrofilariae were assessed by sequential nodulectomies at -3 and 28, 84 and 137 d.p.t. By 137 d.p.t. embryogenesis was almost completely interrupted in the CGP 20376 and ivermectin treated animals, although not in the suramin treated group, but in all three groups the majority of remaining intrauterine microfilariae were pathologically altered. Degenerating intrauterine microfilariae accumulated in the ivermectin and in the CGP 20376, but not in the suramin treated worms. The motility of male and female worms was not reduced by any treatment except for female worms at 84 d.p.t. with CGP 20376. Viability of the worms as indicated by the MTT-formazan reduction assay was not reduced in any of the treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Africa↗