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Biomedical subjects

H J Bürger

Publications and source records attributed to H J Bürger.

At least 19 recordsLinked to original sources

Detection of Trypanosoma congolense antibodies with indirect ELISAs using antigen-precoated microtitre plates.

The study reports the performance of four indirect enzyme-linked immunosorbent assays (ELISAs) for antibody (AB) detection using microtitre plates which were precoated with native or heat/detergent denatured antigens (AGs) from Trypanosoma congolense (T.c.) and T. vivax (T.v.), and stored for between 1 to 206 days at +37 degrees C. Bovine serum samples were obtained by sequential bleeding of 3-months old T.c.-infected bulls and their uninfected cohorts, as well as by a single bleeding of uninfected adult cattle. The first day of AB detection, and observations on samples after this (defined as estimated ELISA sensitivity), depended on the cut-off value in the specific ELISAs. Cut-off values from pre- and early post-infection samples of individual animals demonstrated a seroconversion in all ELISAs on average after 10-15 days post-infection (dpi). The AB detection was delayed in the T.c. native and denatured AG-based ELISAs using cut-off points from uninfected cohort cattle (16.5 dpi, 19.3 dpi) and the adult cattle population (22.1 dpi, 25.0 dpi). The T.v. AG-based ELISAs however lacked crossreactiviy to T.c. ABs. The estimated sensitivity of each T.c. AG-based ELISA was above 96% throughout, but significantly lower for the T.c. native AG-based ELISA (91.1%) when the adult cattle derived cut-off point was used (p<0.01). The sensitivity of the phase contrast buffy coat technique was similar to the T.c. AG-based ELISAs, but significantly lower when the T.c. denatured AG-based ELISA was used at the adult cattle derived cut-off point (p<0.05). The implications of the results and future research aspects on ELISAs to detect trypanosomal ABs and AGs are discussed.

Animals↗

Intracellular calcium and pH conditions of cultured cells infected with Eimeria bovis or E. separata.

Loading of Eimeria bovis-infected Vero cells with membrane-permeant acetoxymethyl esters (AM-esters) of ion-sensitive dyes provided us with a noninvasive method for investigation of the permeability of the parasitophorous vacuole membrane (PVM) and simultaneous measurement of Ca2+ and H+ concentrations in different compartments of the infected cells. The distribution patterns of the cleaved membrane-impermeant dyes argue against the existence of nonselective pores in the PVM. There is also no indication of a parasitophorous duct connecting the vacuolar space with extracellular media. The pH inside the parasitophorous vacuole (PV) was lower than that in the cytoplasm of the host cell or the parasite, whereas the [Ca2+] in these compartments did not differ significantly. In HT29 cells infected with E. separata for 24 h the Ca2+ response to extracellular adenosine triphosphate (ATP) was significantly reduced, indicating influences on the host cell's intracellular signaling.

Adenosine Triphosphate↗

T cell responses in calves to a primary Eimeria bovis infection: phenotypical and functional changes.

The study aimed to characterize T cell responses in calves to a primary E. bovis infection. For this purpose, peripheral blood lymphocytes (PBL) were isolated from six infected calves and three controls during prepatency (Day 12 post infection (p.i.), patency (Day 25 p.i.) and postpatency (Day 35 p.i.). In addition, lymphocytes were isolated from various lymphatic organs (lnn. cervicales superficiales, lnn. jejunales craniales, lnn. jejunales caudales, lnn. caecales, lnn. colici, Peyer's patches (PP) and spleen) at necropsy (Day 35 p.i.). FACS analyses determined the proportions of CD4+-, CD8+-, CD2+-, and gammadelta+-T cells. Proliferative responses of the cells after stimulation with Concanavalin A (Con A) and an E. bovis-merozoite I antigen (EbAg) were measured. Furthermore, in situ hybridization experiments were performed for the detection of IL-2 and IL-4 mRNA in histological sections of lymphatic organs. Proportions of CD4+-, CD8+- and CD2+-expressing PBL were significantly increased 12 days p.i. in infected calves. While the proportions of CD4+- and CD8+-PBL declined until day 25 p.i. and finally reached control values, proportions of activated PBL (CD2+-T cells) remained at a high level throughout the observation period. Those of gammadelta+-PBL, in contrast, remained unaffected. The proportions of CD4+-, gammadelta+- and CD2+-T cells in lymphatic organs were significantly increased in comparison to uninfected controls, when determined 35 days p.i. Concerning the proportions of CD8+-T cells of the organs, however, there were no differences between the groups. PBL and cells from lymphatic organs except those from the PP showed strong proliferative response to the mitogen Con A, without a significant difference between the groups. Reactions to EbAg in contrast differed significantly between controls and E. bovis infected calves. Proliferation responses of PBL of infected animals were highest 12 days p.i.; subsequently they decreased and 35 days p.i. they were found within the ranges of controls. Lymphocytes isolated from lymphatic organs of infected animals reacted significantly stronger than lymphocytes from control animals, whereby most marked differences occured with cells from lymph nodes draining E. bovis infested parts of the intestine and from the spleen. These reactions were accompained by an increased transcription of the IL-2 gene but not of the IL-4 gene in gut associated lymphnodes of infected calves when compared with infected controls. The data suggest strong antigenic stimuli by developing first generation schizonts, and of predominant involvement of (CD4+) Th1 cells in the course of a primary E. bovis infection of calves.

Animals↗

Apparent digestibility of nutrients and nitrogen balance during experimental infection of calves with Eimeria bovis.

The apparent digestibilities (AD) of dry matter (DM), organic matter (OM), crude ash (CA), crude fiber (CFi), crude fat (CFa) crude protein (CP) and nitrogen-free extracts (NFE) and the nitrogen balance were investigated during experimental Eimeria bovis coccidiosis in calves. noninfected pair-fed controls and controls fed on a normal plan of nutrition were included in the study to allow differentiation between the effects of infection and of changes in feed intake. Primary infection with 5 x 10(4) oocysts (n = 4, group A) caused mild diarrhea and calves infected primarily with 1 X 10(5) oocysts (n = 5, group B) suffered from mild (three calves) to severe hemorrhagic (two calves) diarrhea. No clinical disease was seen after reinfection of the group A calves with 1 X 10(5) oocysts. The primary infection with 5 X 10(4) oocysts or reinfection with twice the primary inoculum did not affect AD of nutrients or the overall nitrogen balance (RT). AD of DM, NFE or OM were higher in group B during patency and in the pair-fed group C calves (n = 5) than in the reinfected but healthy group A calves. AD of CFi of the group B calves even exceeded the values of the pair-fed controls. The two calves of group B that suffered from hemorrhagic diarrhea and anorexia had low values of AD of CP during the acute phase of the disease and the plasma nitrogen levels were reduced in this group. Severe clinical coccidiosis transiently reduced the nitrogen balance. It is concluded that the transient increase of AD of nutrients, especially of CFi, during clinical coccidiosis reflect hypomotility and that anorexia and intestinal leakage impair the nitrogen balance and cause weight depression.

Animals↗

Efficacy of two formulations ('injectable' and 'pour on') of moxidectin against gastrointestinal nematode infections in grazing cattle.

The efficacy of moxidectin, 'injectable' and 'pour on', against gastrointestinal nematodes was determined in cattle in two separate field trials (Trial I in 1990 and Trial II in 1991) with respectively 88 and 94 young grazing cattle of either sex. The efficacy was measured on the basis of the reduction of the egg output and of the evaluation of the results from larval differentiation. Animals in Group MI received 0.2 mg kg-1 body weight (b.w.) moxidectin injectable solution in Trial I on Day 0. Group CI was not given any medication on Day 0, but 0.2 mg kg-1 b.w. ivermectin injectable solution (Ivomec) was applied after 2 weeks to prevent clinical disease. In Trial II, animals in Group MP were treated with pour on moxidectin (0.5 mg kg-1 b.w.) on Day 0. Animals in Group CP serving as controls for Group MP during the first part of the trial received the same formulation at the same dose 2 weeks after treatment of Group MP. When the egg output was compared within treated groups, the egg count reduction was very similar post treatment (p.t.) with both formulations being 96.3% and 96.6% on Day 7 after the application of injectable moxidectin or pour on moxidectin, respectively, and 90.7% and 92.5% on Day 28 p.t. When egg counts of treated and control animals were compared (corrected for the e.p.g. values before treatment) the egg count reduction was 95.4% and 91.5% on Day 7 and 92.9% and 84.8% on Day 14 p.t. with either the injectable or pour on formulation. Pour on moxidectin seemed to be more effective against Ostertagia spp. than against Cooperia spp. Animals treated with injectable moxidectin gained significantly more body weight (4.2 kg per animal) than the controls from Day -7 to Day +14, while no significant difference in weight gain was achieved within 2 weeks after treatment with pour on moxidectin.

Administration, Topical↗

Eimeria bovis in cattle: colostral transfer of antibodies and immune response to experimental infections.

IgM, IgG1, and IgG2 antibodies to Eimeria bovis first-generation merozoite antigens were determined by enzyme-linked immunosorbent assay and Western blotting in naturally infected cows and in their offspring before and after the uptake of colostrum. In addition, calves were examined following experimental primary and challenge infections. Neonate calves received maternal antibodies via colostrum. All isotypes determined were transmitted, but only IgG1 was concentrated in the colostrum and it occurred at significantly increased levels in sera from the calves as compared with those from the respective dams. Recognition patterns (Western blotting) displayed by related maternal serum and colostrum and those shown by calves that had ingested colostrum were very similar, but marked variations occurred between individual pairs. Experimental infection of 15-week-old calves with 0.7 x 10(5) oocysts caused strong protective immunity against a challenge with 1 x 10(5) oocysts. In contrast, animals that had undergone a weak intercurrent infection were not protected. Experimental infections induced a considerable increase in IgG1 and IgG2 antibody levels, whereas IgM values increased only slightly. The spectrum of merozoite antigens recognized by the sera increased markedly after experimental infection, although high individual variations were found in the calves. However, there was no correlation between the levels of any specific antibody or the recognition patterns and the status of immunity to a severe challenge.

Animals↗

Prevalence of gastro-intestinal nematode infection in the dromedary camel (Camelus dromedarius) in the Butana plains, Sudan.

The prevalence and intensity of gastro-intestinal nematode infection in their relation to season and rainfall were investigated from 429 female dromedary camels at Tambul market in the Butana plains (Sudan), during 1985-1986. The investigation revealed a similar seasonal pattern in the prevalence as well as the intensity of egg output. The seasonality is mainly brought about by Haemonchus spp. and Impalaia spp. while Trichostrongylus spp. seem to be present as adults throughout the year. There is a good correlation between high egg counts and rainfall ensuring optimal development of preparasitic stages.

Animals↗

Efficacy of four anthelmintics against benzimidazole-resistant cyathostomes of horses.

In order to confirm benzimidazole resistance as recommended at a workshop of the Commission of the European Communities the isolate 'E' of cyathostome strongyles originating from a stud where benzimidazole resistance had been demonstrated by egg hatch tests and by egg count reduction tests was investigated in two series of critical tests. Each of 11 foals reared strongyle-free was infected with 130,000 third stage cyathostome larvae. One animal remained untreated, two pairs of foals were treated with paste formulations of the (pro)benzimidazoles cambendazole (20 mg/kg bodyweight) or febantel (6 mg/kg bodyweight) and two groups of three foals were given pastes containing the non-benzimidazole drugs pyrantel pamoate (19 mg/kg bodyweight) or ivermectin (0.2 mg/kg bodyweight) either at 101 days (trial 1) or at 59 to 62 days (trial 2) after infection. Strongyles were counted in faecal samples collected daily between treatment and post mortem examination five or seven days later and worm burdens were counted in the intestinal contents and mucosal digests. Nine species of the cyathostome subfamily were found in the infected foals. The numbers of luminal stages were reduced by only 3.1 and 20.2 (mean 7.9) per cent by cambendazole and by 13.6 and 32.8 (mean 21.3) per cent by febantel in the individual animals. However, pyrantel pamoate (93.6 to 98.2, mean 96.3 per cent reduction) and ivermectin (100 per cent reduction) were highly effective. These trials provide the first report of benzimidazole resistant Cylicostephanus poculatus anywhere in the world and demonstrate (pro)benzimidazole resistance in seven other species for the first time in Europe.

Animals↗

Efficacy of an oxibendazole-trichlorfon paste formulation against third stage larvae of Gasterophilus intestinalis and its safety in horses.

A paste formulation containing 14.3 per cent of oxibendazole and 44 per cent of trichlorfon was administered to 33 ponies and horses. The dose rate used was equivalent to 10 mg and 30 mg/kg bodyweight, of oxibendazole and trichlorfon respectively. After treatment 25 animals passed between one and 82 third stage larvae of Gasterophilus intestinalis in their faeces. Dosing with 0.2 mg ivermectin/kg bodyweight three weeks later resulted in six animals expelling between one and four bots. The efficacy of the oxibendazole-trichlorfon paste was on average 96.2 per cent. This drug combination given to 52 ponies and horses at the indicated dose rate and to six ponies at twice that dose was tolerated without side effects except transient softening of the faeces in several animals and mild symptoms of colic in two horses.

Administration, Oral↗

Prevalence and control of benzimidazole-resistant small strongyles on German thoroughbred studs.

The prevalence of benzimidazole-resistant small strongyles was determined in a survey, conducted on 14 thoroughbred studs, which compared the faecal egg counts of groups of horses before and after treatment with the recommended doses of cambendazole (20 mg kg-1 b.w.) or febantel (6 mg kg-1 b.w.). Benzimidazole-resistant cyathostomes were found on all farms examined. Pyrantel pamoate (19 mg kg-1 b.w.), oxibendazole (10 mg kg-1 b.w.) and ivermectin (0.2 mg kg-1 b.w.) reduced the strongyle egg counts on these studs by 97-100% at 2 weeks post-treatment. However, 6 weeks after dosing the reduction of the strongyle egg output had decreased to an average of 67.8% (8.7-97.1%) with pyrantel pamoate and 51.2% (0-95.8%) with oxibendazole, whereas ivermectin still suppressed the egg counts by 98.2% (95-100%).

Animals↗