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.     

Lymphocyte responses to mitogens and rickettsial antigens in sheep experimentally infected with Ehrlichia (Cytoecetes) phagocytophila.

Infection of sheep with Ehrlichia (Cytoecetes) phagocytophila, the causative agent of tick-borne fever (TBF), was characterised by a significant reduction in lymphocyte reactivity to the mitogens phytohaemagglutinin, concanavalin A, pokeweed mitogen and Escherichia coli lipopolysaccharide during the period of rickettsiaemia. The addition of the prostaglandin inhibitor, indomethacin, or the nitric oxide inhibitor, N(G)-monomethyl-L-arginine, had no significant effect on the suppressive effects of E. phagocytophila on lymphocyte reactivity to the mitogens. However, peripheral blood lymphocytes obtained from primed sheep proliferated in the presence of live or heat-inactivated E. phagocytophila. Antigen-specific proliferation was detected in lymphocytes samples obtained 11 to 21 days post-inoculation with E. phagocytophila.     

Detection of antibodies to the agent of tick-borne fever by indirect immunofluorescence

Cytospin preparations of sheep granulocytes infected with Cytoecetes phagocytophila, the causative agent of tick-borne fever (TBF), enabled the development of an improved and reproducible indirect fluorescent antibody test to detect antibodies in the sera of sheep convalescing from TBF. The test was compared with counter-immunoelectrophoresis.     

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.     

Clinical, haematological and blood biochemical changes in goats after experimental infection with tick-borne fever

Tick-borne fever in goats caused by Ehrlichia (Cytoecetes) phagocytophila was characterised by high fever, dullness, anorexia, tachycardia and a slight to moderate inhibition of rumen motility. The animals developed a gradual decline in the total number of circulating white blood cells. There was a decrease in lymphocytes over a short period, followed by an increase. The number of neutrophils was higher on the 3rd day, causing considerable change in the lymphocyte:neutrophil ratios. The number of eosinophils increased slightly. Serum alkaline phosphatase (ALP) decreased during the febrile episodes, and a marked decline was observed in both plasma zinc and iron concentrations. Furthermore, there was a small but progressive decrease of haemoglobin and haematocrit values. Circulating endogenous pyrogen/leucocyte endogenous mediator could not be detected in plasma from febrile goats. Tick-borne fever was passively transmitted to kids with plasma obtained from these febrile animals.     

Immune evasion and immunosuppression by Anaplasma phagocytophilum, the causative agent of tick-borne fever of ruminants and human granulocytic anaplasmosis

Anaplasma phagocytophilum, the causative agent of tick-borne fever (TBF) in sheep and cattle and human granulocytic anaplasmosis, has the unique ability to infect and multiply within neutrophils, eosinophils and monocytes, cells at the frontline of the immune system. Infection with A. phagocytophilum is also characterized by severe leukopenia due to lymphocytopenia, neutropenia and thrombocytopenia lasting for several days. By itself TBF does not cause high mortality rates but infected animals are more susceptible to other secondary infections, pregnant animals may abort and there is a severe reduction in milk yield in dairy cattle. The susceptibility to secondary infections can be attributed to the leukopenia that accompanies the disease and the organism's adverse effects on lymphocyte and neutrophil functions. One of its fascinating features is that it infects and actively grows in neutrophils by employing an array of mechanisms to subvert their bactericidal activity. These include its ability to inhibit phagosome-lysosome fusion, to suppress respiratory burst and to delay the apoptotic death of neutrophils. It is also able to survive within an apparently immune host by employing a complex mechanism of antigenic variation.     

Resistance to tick-borne fever in young lambs.

Lambs were inoculated with Ehrlichia phagocytophila infected blood at the age of either two or six weeks. The temperature response and degree of parasitaemia were both most pronounced in the lambs inoculated at six weeks old. The half-life of antibodies to E phagocytophila was estimated to be 17.5 days. Lambs inoculated with tick-borne fever (TBF) had lower liveweights in the autumn than TBF-free controls.     

Use of counter immunoelectrophoresis in detection of antibodies to tickborne fever

A micromethod using the counter immunoelectrophoresis technique for the detection of antibodies to Cytoecetes phagocytophila, the causative agent of tickborne fever (TBF) is described. Antibodies were first detected nine to 11 days after experimental infection of lambs with TBF and persisted for six to 10 weeks after infection. The test was also applied to approximately 440 field samples of ovine sera collected from tick infested farms in Scotland and the north of England, 16 per cent of which proved positive and showed a marked seasonal variation.     

Congenital infection with Anaplasma phagocytophilum in a calf in northern Germany

Anaplasma phagocytophilum is a Gram-negative, obligate intracellular tick-transmitted bacterium that replicates in neutrophils. It causes tick-borne fever (TBF) in sheep and cattle, but also elicits febrile disease in humans as well as in other domestic animals such as dogs, horses, and cats. Although increasingly recognized in Europe, the first laboratory-confirmed case of TBF in cattle from Germany has been published only recently. We here present the unusual case of an intrauterine transmission of A. phagocytophilum in a calf from northern Germany. To our knowledge, this is the first report of such an event occurring under field conditions in cattle.     

Lymphocyte subpopulations in peripheral blood of sheep experimentally infected with tick-borne fever.

The lymphocyte subpopulations in the peripheral blood of normal sheep and sheep experimentally infected with Cytoecetes phagocytophila, the causative agent of tick-borne fever, were analysed by flow cytometry, using a panel of monoclonal antibodies against specific lymphocyte epitopes. Experimental infection with tick-borne fever was characterised by a significant reduction in the total number of circulating lymphocytes six days after experimental infection (P less than 0.001). This lymphocytopenia was associated with a significant reduction in the number of B (LCAp220+) and T (CD5+) lymphocytes (P less than 0.001) but there was a significant increase in the number of cells which were neither T nor B (CD5-LCAp220-) cells (P less than 0.01). The reduction in the number of T lymphocytes was due to reduced numbers of circulating CD4+ (helper) T cells, CD8+ (cytotoxic/suppressor) T cells and those with the pan T cell marker (CD5+) but without CD4 or CD8 epitopes (CD4-CD8-). All lymphocytes returned to preinoculation levels 13 to 16 days after experimental infection.     

Effect of tick-borne fever on the disposition of sulphadimidine and its metabolites in plasma of goats

The effect of tick-borne fever (TBF) on the plasma disposition of sulphadimidine (SDM) and its metabolites in goats was studied. In uninfected goats, SDM was extensively metabolised mainly by hydroxylation, glucuronidation and to a minor extent by acetylation. In TBF infected goats the hydroxylation of SDM into 6-methylhydroxysulphadimidine (SCH2OH) as well as into 5-hydroxysulphadimidine (SOH) was markedly reduced (-57.6 and -63.6 per cent, respectively). An unidentified metabolite (metabolite X) was detected, which was largely glucuronidated in the uninfected goats. In the TBF infected goats the glucuronide derivatives of the X metabolite and of SOH were barely detectable. In TBF infected goats the plasma concentration of the N4-acetylated metabolite (N4-SDM) was decreased to a lesser extent (-22.1 per cent) than the hydroxy metabolites. Due to the diminished metabolism the elimination half-life of SDM was increased 1.8 times and the total sulphonamide body clearance was diminished compared with findings in the control experiments.     

Q fever serologic surveillance program for sheep and goats at a research animal facility

Since Q fever is a potential risk to personnel working with small ruminants, the serologic status of sheep and goats received at a medical school animal facility for research was evaluated. A total of 104 sheep and 102 goats were subjected to blood sample-collection procedures on arrival, as well as after a 2-week quarantine period, and the sera were tested for Q fever specific antibodies by complement-fixation (CF) and microagglutination (MA) tests. The results from the 2 tests were compared and analyzed for seroconversion. On the basis of the CF test, 14 sheep and 3 goats were considered positive; these included 7 sheep and 2 goats that seroconverted during the quarantine period. In contrast, 1 sheep and 5 goats were found positive by the MA test, which also detected seroconversion of 1 sheep and 1 goat. The use of both tests for serologic surveillance of Q fever in sheep and goats increased the likelihood of detection. Management safety practices are also required to minimize the risk of disease transmission.     

Anaplasma phagocytophilum - the most widespread tick-borne infection in animals in Europe

The bacterium Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila) may cause infection in several animal species including human. The disease in domestic ruminants is also called tick-borne fever (TBF), and has been known for at least 200 years. In Europe, clinical manifestations due to A. phagocytophilum have been recorded in sheep, goat, cattle, horse, dog, cat, roe deer, reindeer and human. However, seropositive and PCR-positive mammalian have been detected in several other species. Investigations indicate that the infection is prevalent in Ixodes ricinus areas in most countries in Europe. A. phagocytophilum infection may cause high fever, cytoplasmatic inclusions in phagocytes and severe neutropenia, but is seldom fatal unless complicated by other infections. Complications may include abortions, and impaired spermatogenesis for several months. However, the most important aspect of the infection at least in sheep is its implication as a predisposing factor for other infections. Factors such as climate, management, other infections, individual conditions etc. are important for the outcome of the infection. A. phagocytophilum may cause persistent infection in several species. Based on the 16S rRNA gene sequences several variants exist. Different variants may exist within the same herd and even simultaneously in the same animal. Variants may behave differently and interact in the mammalian host.     

An electron microscopic study of Cytoecetes phagocytophila infection in Ixodes ricinus

Tick-borne fever, caused by a rickettsia-like organism, Cytoecetes phagocytophila, is transmitted by the sheep tick Ixodes ricinus. An electron microscopic technique was developed to examine I ricinus for C phagocytophila infection. Infected and uninfected ticks were obtained from a laboratory maintained tick culture. All stages of I ricinus collected from one field site were examined; 44 per cent of nymphae and 32 per cent of adults were infected with C phagocytophila, but larvae were uninfected. This supports the previously held theory of transtadial, but not transovarial transmission of tick-borne fever.     

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.     

Caprine hepatic lipidosis induced through the intake of low levels of dietary cobalt

Forty-one, 10-week-old newly weaned goats were randomly allocated into two groups, namely control (n=22) and treated (n=19). Kids in both groups were fed Rhodegrass hay ad libitum that contained < 0.1 mg/kg DM cobalt and 150 g/day of a commercially prepared ruminant concentrate that contained approximately 0.12 mg/kg DM cobalt. This diet provided the minimum daily requirement of cobalt as specified for sheep. The treated goats were supplemented with bi-monthly subcutaneous injections of 2000 microg of hydroxycobalamin. All goats were weighed and blood samples collected monthly for haematological, clinical biochemical and serum vitamin B12 analysis. After a 10-month experimental period the goats were slaughtered. The control animals exhibited significantly (P<0.05) lower weight gains, and had dry scruffy hair coats. In addition, there was a decline in erythrocyte counts, mean haemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Controls also exhibited significantly (P<0.05) lower levels of total serum proteins and elevated levels of serum alkaline phosphatase compared to treated goats. Fourteen (63.6%) of the control goats developed pathology consistent with reported field cases of hepatic lipidosis associated with low liver levels of cobalt. Only one (5.3%) of the treated goats developed hepatic lipidosis. Contrary to previous reports that suggested that goats are less sensitive to low levels of dietary cobalt than sheep, it is apparent that this is not the case with Omani goats. This is the first report of the induction of hepatic lipidosis in goats due to feeding low levels of cobalt in their diet.     

Peste des petitis ruminants morbillivirus infection in lambs and young goats at Qassim region, Saudi Arabia

An outbreak of peste des petits ruminants (PPR) in lambs and young goats of Najdi breed of sheep and goats occurred during winter 2005 at Qassim region of central Saudi Arabia. The PPR infection was confirmed by demonstration of antibodies against the virus in the serum of clinically-ill young sheep and goats using competitive ELISA test. Clinical examination of infected animals showed fever, salivation, lacrimation, mucopurulent nasal discharge, difficult breathing and diarrhoea. The disease was particularly severe in the goats. Morbidity was about 20% and mortality was less than 3 percent. Autopsy showed necrotic and ulcerative lesions in the mouth, stomach and intestine. Mesenteric lymph nodes were swollen and congested. The lungs were patchy pneumonic mostly at the diaphragmatic and apical lobes. Liver and kidney lesions were seen in goats only and both organs were congested and necrotic. Histopathological examination revealed necrotic inflammation in the gastrointestinal tract. Intracytoplasmic viral inclusions were seen in the enterocytes of goats. Lung sections showed bronchopneumonia and syncytial and giant cells. The bronchial epithelium of goats had intracytoplasmic viral inclusions. Extensive coagulation necrosis, fatty degeneration and presence of intracytoplasmic viral inclusions were seen in hepatocytes and syncytial cells were evident in biliary epithelium of goats. Congestion, coagulation necrosis and syncytial cells were seen in the renal tubular epithelium of goats only. In a survey to determine the magnitude of the outbreak, PPR antibodies were evidenced in 363/996 (36.6%) sheep and 530/962 (55.1%) goats.     

Goat smallpox in Chad: study of the pathogeny of the virus in sheep and goats]

A local strain of goat pox virus was tested in goats and sheep. The results showed that 65% of goats and 20% of sheep reacted positively. Only goats died few days after the inoculation (55%) and no mortality was recorded in the sheep. The strict species specificity of this strain was not observed. The difference of sensitivity between sheep and goats was statistically significant.     

Listeric septicaemia in sheep associated with Tick-Borne fever (Ehrlichiosis ovis).

Sheep is apparently fairly resistant against infections with Listeria monocytogenes (Lm). Grønstøl (unpublished) found that a high proportion of healthy sheep are carriers of Lm. In such animals Lm may cause clinical disease when resistance is lowered. Tick-borne fever (TBF) is known to reduce the resistance against several infections (Øverås 1972). This report describes listeric septicaemia associated with TBF.     

Clinical and Para-clinical Findings of a Recent Outbreaks of Peste des Petits Ruminants in Iran

Peste des petits ruminants (PPR) is a highly contagious and infectious viral disease of domestic and wild small ruminants characterized by fever, erosive stomatitis, conjunctivitis, gastroenteritis and pneumonia. Goats are usually more severely affected than sheep. Peste des petits ruminants is caused by a paramyxovirus of the Morbillivirus genus. In March 2004, a flock of sheep in Tehran province with 430 deaths was visited. According to the history taken from the owner, at disease onset most of the deaths were recorded from adult sheep, 3 weeks later lambs (2 weeks to 4 months of age) showed the highest death rate. All animals from 3 months age received rinderpest vaccine 1 month after onset. Many of the lambs died just a few hours after their first sucking of the colostrum from infected mothers. Most of them showed very acute form of disease and died a few hours after onset of clinical signs. In clinical examinations most of the cases showed severe depression, high fever (41 degrees C), anorexia, mocopulurent nasal discharge, erosive and necrotic stomatitis (dental path, hard palate and cheeks), diarrhoea and dehydration. Para-clinical findings including histopathological, serological and haematological examinations also confirmed the presence of PPR in this flock. PPR outbreaks have been frequent in Iran in recent years. Further, we suggest that PPR is not a recent invader of Iran. The main difference in clinical signs between this outbreak and the same in other reports is that goats did not show any obvious signs of PPR. This might be due to the number of the goats (>1% of the flock) and keeping them separate from the sheep. The present article reviews the details of this outbreak in Iran.