The effect of two different oxytetracycline treatments in experimental Ehrlichia phagocytophila infected lambs

The effect of 2 different oxytetracycline treatments in acute E. phagocytophila infected lambs was investigated. Twenty 5-month-old lambs of the Dala and Rygja breeds were used. Ten lambs were inoculated intravenously with a stabilate of an ovine E. phagocytophila strain. On the third day of fever, 4 lambs were given long-acting oxytetracycline (Terramycin prolongatum vet, Pfizer) (20 mg/kg) intramuscularly and another 4 lambs were given short-acting oxytetracycline (Terramycin vet, Pfizer) (10 mg/kg) intravenously for 5 consecutive days. The lambs were examined for the presence of Ehrlichia infection by blood smear evaluation, polymerase chain reaction (PCR) and antibody titre against E. equi. One month after the last antibiotic treatment, 250 ml citrate blood from each of these lambs were inoculated into each of 10 susceptible lambs, which were observed during the following 6 weeks. The results indicate that oxytetracycline given in the acute stage of the infection may effectively terminate the development of fever, rickettsemia and weight reduction in E. phagocytophila infected lambs. No difference was observed between the 2 treatment groups. However, at least 3 of 8 antibiotic treated lambs (37.5%) were still infected with granulocytic Ehrlichia 3 months after treatment.     

Persistence of Ehrlichia phagocytophila infection in two age groups of lambs

Tick-borne fever (TBF) is caused by the rickettsiae Ehrlichia phagocytophila and is a common disease in sheep in tick (Ixodes ricinus) infested areas in Norway. Earlier investigations have shown that some sheep could remain infected for several months after the primary infection. In this study, the persistence of E. phagocytophila after experimental infection was investigated in 2 age groups of lambs. Six lambs (1-2 weeks old) and 14 lambs (6-8 months old) were inoculated intravenously with an ovine strain of E. phagocytophila and thereafter examined clinically (including daily body temperature recording) and by haematological and serological (E. equi antibodies) methods for the next 4 months. At the end of this period, the lambs were examined for a TBF infection by blood smear investigation and blood inoculation studies. The infection was demonstrated in 19 (95%) of the 20 lambs.     

Attempts to cultivate Ehrlichia phagocytophila in vitro

Whole blood, buffy coat and plasma from sheep infected with tick-borne fever (TBF) were inoculated into embryonated hen eggs, primary chicken embryo fibroblasts, primary foetal lamb kidney cells, mouse L cells, and ovine pulmonary macrophage cultures. Infected buffy coat cells were fused with primary chicken embryo fibroblasts, mouse L cells, ovine pulmonary macrophages and BHK-21 cells, using UV irradiated Sendai virus. Evidence of multiplication was not obtained.     

Functions of neutrophils in sheep experimentally infected with Ehrlichia phagocytophila

Ehrlichia phagocytophila infection in sheep is characterized by persistent neutropaenia, indicative of decreased phagocytic capacity. This predisposes infected animals to other infections. A whole blood flow cytometrical method was used to document the degree and extent of reduced phagocytic and respiratory burst activity in phagocytes during an experimental infection with E. phagocytophila, and monitored until 56 days post-infection. Six sheep at 5 months of age were inoculated with an intravenous injection of infected blood. Six age-matched sheep were used as controls. A period of reduced respiratory burst lasting up to Day 17 post-infection was recorded. The population of cells showing phagocytic activity without respiratory burst was larger in the infected animals compared to controls up to Day 45 post-infection.     

Lack of cross-protection between Cowdria ruminantium and Ehrlichia phagocytophila.

Antigenically distinct stocks of Cowdria ruminatium from Senegal and South Africa were compared with a Dutch isolate of Ehrlichia phagocytophila in cross-immunity trials in goats. There was a complete absence of cross-immunity between E. phagocytophila and C. ruminantium, despite previous observations that both rickettsial organisms have certain antigenic determinants in common.     

Tick-borne fever (Ehrlichia phagocytophila infection) on a dairy farm in Friesland

Disease outbreaks, associated with ixodid ticks, among cows on a dairy farm in Friesland, in the north of the Netherlands, were monitored during the summers of 1996, 1997, and 1998. The most important symptoms were a sudden drop in milk yield, fever, apathy, and problems with locomotion. The diagnosis 'tick-borne fever' was confirmed by finding Ehrlichia phagocytophila inclusion bodies in neutrophils of blood smears from sick animals. Significant economic losses due to the disease were recorded each year. Borrelia burgdorferi, although transmitted by the same tick, did not seem to be involved. Further research on tick-borne fever is of particular relevance in connection with recent E. phagocytophila cases diagnosed in dogs and in a human patient in the Netherlands.     

Effect of monensin on experimental infections of Eimeria ninakohlyakimovae in lambs.

Two nearly identical experiments were conducted to determine the effects of the coccidiostatic compound monensin on Polled Dorset lambs experimentally infected with oocysts of Eimeria ninakohlyakimovae. Prophylactic medication at a dose level of 1 mg/kg of body weight was started 2 days before inoculation and prevented diarrhea and reduced oocyst production. Therapeutic medication at a dose level of 2 mg/kg, started with the appearance of signs of infection, reduced oocyst production below that of nonmedicated controls but did not eliminate diarrhea. Weight gains of medicated lambs were less than those of noninoculated, nonmedicated controls in both experiments. Apparently monensin, at the dose levels used, reduced oocyst production but also prevented weight gains as high as those in inoculated nonmedicated controls.     

Natural and experimental infections of lambs with Mycoplasma ovipneumoniae

Mycoplasma (M.) ovipneumoniae was isolated pure or mixed with bacteria from 47 lungs of lambs of 14 in 22 tested flocks. M. ovipneumoniae was obtained as pure culture in cases of mild bronchopneumonia. Experimental intratracheal or intranasal infection caused several days of rising body temperature above 39.7 degrees C. Nasal discharge, coughing, and dyspnea did not occur. M. ovipneumoniae was successfully re-isolated from nasal swabs, beginning 2 d from infection. Lobular catarrhal bronchopneumonia was established by postmortem examinations, 10-14 d from infection, and M. ovipneumoniae was re-isolated from the lungs. Histological patterns of lungs were characterised by interstitial cell reactions.     

Effects of Ehrlichia phagocytophila infection on serum thyroid hormone concentrations and on antipyrine clearance and metabolite formation in dwarf goats.

The influence of infection with Ehrlichia phagocytophila (EP) on serum thyroid hormone concentrations and on antipyrine (25 mg/kg of body weight, IV) plasma elimination and urinary metabolite excretion was studied in castrated male dwarf goats. Mean thyroid hormone concentrations moderately decreased in EP-infected goats, with maximal decrease in total and free triiodothyronine and thyroxine serum concentrations of 56, 64, 23, and 19%, respectively. The estimated pharmacokinetic values of antipyrine (AP) in EP-infected goats were similar to those in the goats when healthy. However, glucuronidation of the AP-metabolites 3-hydroxymethyl-AP, 4,4'-dihydroxy-AP, and 4-hydroxy-AP was reduced during the febrile episode of the acute-phase response to EP infection.     

The effects of Ehrlichia (Cytoecetes) phagocytophila on the clinical chemistry of sheep and goats

Tick-borne fever (TBF) is a rickettsial disease of domestic and wild ruminants in temperate climates where the hard tick Ixodes ricinus is present. The disease is characterized by a high temperature and severe leukopenia. In the present study, the effects of TBF on the activity of serum alkaline phosphatase (ALP), and on the concentrations of plasma zinc, iron, total bilirubin, urea, creatinine and albumin were investigated by inoculating one group of eight sheep and one group of eight goats with the Old Sourhope (OS) strain of Ehrlichia (Cytoecetes) phagocytophila. All goats and sheep experimentally infected with E. phagocytophila reacted with fever, rickettsiaemia and leukopenia. The leukopenia was due to an acute lymphocytopenia and prolonged neutropenia. In both groups of animals. TBF was characterized by significant reductions in the activities of serum ALP and concentrations of plasma zinc, iron and albumin. However, there were significant increases in the concentrations of plasma total bilirubin, urea and creatinine in both species of animals. The reductions in ALP and iron were significantly more pronounced in sheep than in goats.     

A modification of the indirect immunofluorescence test for detection of Ehrlichia phagocytophila antibodies.

A modified technique for production of antigen and performance of the test is described. A suspension of infected neutrophils was directly applied to multiwell slides. Multichannel pipettes may be used for dilution and application of sera. The modification increases the capacity both by production of the antigen and by performance of the test. This paper also gives a quantitative determination of the antibodies.     

Serotypes in Cowdria ruminantium and their relationship with Ehrlichia phagocytophila determined by immunofluorescence

Two tick-borne rickettsial pathogens of ruminants, Cowdria ruminantium (causative agent of heartwater disease) and Ehrlichia phagocytophila (causative agent of tick-borne fever), were successfully cultivated in caprine or ovine neutrophilic granulocytes. Infected cultures were subsequently used as antigens in the indirect fluorescent antibody test. Low-level bilateral serological cross-reactions could be detected between Cowdria and Ehrlichia. In addition, comparison of five Cowdria stocks using immunofluorescence demonstrated the existence of distinct serotypes within the genus of Cowdria. It is concluded that the occurrence of these serotypes will considerably complicate the current serodiagnosis of heartwater.     

Experimental cross-infections with Ehrlichia phagocytophila and human granulocytic ehrlichia-like agent in cows and horses.

Four cows and four horses were infected experimentally with Ehrlichia phagocytophila, the cause of tickborne fever in ruminants, and with human granulocytic ehrlichia-like agent, a recently discovered species that infects people, horses and dogs in the USA and Europe. They were infected in either order, 30 days apart, to investigate serological cross-reactivity within the Ephagocytophila genogroup. The course of infection was assessed by routine clinical, haematological, serological and polymerase chain reaction (PCR) examinations. Two of the cows infected with Ephagocytophila and two of the horses infected with granulocytic ehrlichia-like agent, developed characteristic signs of ehrlichiosis. When the same animals were infected with their heterologous ehrlichial isolate 30 days later, they did not develop clinical signs of disease. The infection of the other two cows with human granulocytic ehrlichia-like agent and the other two horses with Ephagocytophila, resulted in asymptomatic seroconversion. When the same animals were infected with their homologous ehrlichial isolate 30 days later, they remained asymptomatic and had normal haematological results and negative PCRS until the end of the monitoring period, 60 days after the first infection. In these animals, there was an increase in antibody titre after the second infection which was interpreted as a specific immune response, and as a reactivation of the immune response to the first infection.