Suspected Lyme borreliosis in sheep.

Two cases of suspected Lyme borreliosis in lambs are described. The clinical signs observed resemble those described for Lyme borreliosis in dogs and horses. The lambs were approximately six months old and came from two different flocks in a district heavily infested with Ixodes ricinus ticks. The lambs had pronounced lameness in one leg, generally bad condition and poor appetite. They had high serum levels of IgG antibodies to Borrelia burgdorferi when tested in an ELISA. The other lambs in the flocks were negative to B burgdorferi antibodies. Attempts were made to isolate spirochaetes from one of the lambs, but were unsuccessful.     

Serological investigation of granulocytic Ehrlichia infection in sheep in Norway

Serum samples of 749 sheep from 75 sheep flocks in Norway, i.e. 361 lambs (6 to 7 months old) and 388 adults (>1.5 year), were analysed for antibodies to Ehrlichia equi. Ten animals from each flock were examined. Seropositive animals were found along the coast of southern Norway from Vestfold to Sør-Trøndelag (as far north as 63°38''N). Seropositive sheep were not found in southeast, east or northern Norway. Thirty-two flocks were seropositive, although tick-borne fever had only been diagnosed earlier in half of these. In 78% of the seropositive flocks, more than 80% of the sheep were seropositive. A total of 35.7 % and 36.3 % of lambs and adults were found seropositive, respectively. However, the overall seroprevalence among animals that had been grazing on Ixodes pastures were 0.80 for the lambs and 0.84 for the adults. Mean antibody titres (± SD) (log₁₀) in seropositive lambs and adults were 2.59 (± 0.449) and 2.70 (± 0.481), respectively. No significant differences in either seroprevalence or mean antibody titre between sheep of different ages were obtained in this study. Based on antibodies 94% of sheep flocks on Ixodes pastures were infected with a granulocytic Ehrlichia infection. The association between seropositive flocks and Ixodes infested pasture shows a very high degree of agreement (p < 0.00001). The present study indicates that granulocytic Ehrlichia infection in sheep is underdiagnosed in Norway.     

Tick infestation and the prevalence of Borrelia burgdorferi and Babesia divergens in cattle in Bavaria

During the grazing period 2002 319 cattle from 31 farms located in 6 districts of southern Bavaria were examined for the presence of ticks in 4- to 5-week intervals, and 287 serum samples were tested for the presence of antibodies against Borrelia burgdorferi and Babesia divergens. Ticks were detected in all 31 farms with a mean prevalence of 69%. 3218 out of 3453 collected ticks were Ixodes ricinus; 139 nymphs, 19 larvae and 77 damaged adult specimens could only be determined to the Genus level (Ixodes).The seasonal pattern revealed the highest frequencies of ticks in May/June and September.The intensity of tick infestation of positive animals was generally low. 76.5% of parasitized cattle had 1-6 ticks per day of investigation. Individual cattle showed up to 250 ticks per day. The percentage of infested animals in each herd varied within the period between 0-100%. The examination of serum samples by immunofluorescence technique (IFAT) revealed positive anti-Borrelia antibody titers (> or = 1:64) for 45.6% of the animals. The within-farm seroprevalence of borreliosis ranged from 20 to 100% in 27 of the 31 farms. A significant correlation could be detected between the number of ticks/cattle and the anti-Borrelia burgdorferi IgG-titer. By contrast, there was no significant correlation between the age of the animals and anti-Borrelia serum titers. For comparative reasons, 64 IFAT-positive serum samples were tested by Western blot techniques for the presence of antibodies cross-reacting with Borrelia garinii antigen. These analyses revealed that 69% of the samples reacted positively, 28% were unclear and 3% were negative. Examinations of the 287 serum samples for the presence of anti-Babesia divergens antibodies revealed one positive animal with a titer of 1:16.     

Vitamin E status of healthy Swedish cattle

Using high pressure liquid chromatography the serum concentration of vitamin E was measured in dairy cows fed either hay or silage as their main roughage, in calves fed milk-replacer, and in young intensively fed bulls. The concentrates fed to the cows, calves and bulls were supplemented with 5–10, 25 and 5–10 mg DL-α-tocopheryl acetate per kg, respectively, and the milk-replacer for the calves was supplemented with 50 mg DL-α-tooopheryl acetate per kg powder. Cows fed silage as their main roughage had higher serum vitamin E concentrations (: 3.8–5.2 mg/l) than cows fed only hay (: 2.5–4.1 mg/1). Lactating cows had higher vitamin E concentrations than dry cows (: 4.1–5.2 and 2.5–3.8 mg/l, respectively) and calves and bulls had much lower vitamin E concentrations (: 1.4 and 1.2 mg/l, respectively) than cows. Thirty per cent of the calves and 41 % of the bulls had serum vitamin E concentrations less than 1.0 mg/l, suggesting that in these animals the conventional level of supplementation of feeds with DL-α-tocopheryl acetate in Sweden is probably inadequate for the prevention of nutritional muscular degeneration and other negative effects.     

Evidence of Bartonella sp. in questing adult and nymphal Ixodes ricinus ticks from France and co-infection with Borrelia burgdorferi sensu lato and Babesia sp

Ticks are known vectors for a wide range of pathogenic microorganisms. Their role in the transmission of some others is so far only suspected. Ticks can transmit multiple pathogens, however, little is known about the co-existence of these pathogens within questing ticks. We looked for the presence of DNA from three micro-organisms, Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. which are known or suspected tick-borne pathogens, using a cohort of 92 questing Ixodes ricinus ticks collected from pastures in northern France. DNA was extracted from each individual tick and the presence of the three pathogens was investigated using Polymerase Chain Reaction (PCR) amplification. Nine among 92 samples (9.8%) demonstrated PCR products using Bartonella specific primers, 3 among 92 (3.3%) using Borrelia burgdorferi sensu lato specific primers and 19 among 92 (20.6%) using Babesia specific primers. Seven among 92 samples (7.6%) were PCR positive for at least two of the pathogens and one sample was positive for all three. Adult ticks (12/18; 67%) showed significantly higher infection rates compared to nymphs (11/74; 15%) for all three pathogens (P < 0.001). This study is the demonstration of the simultaneous presence of Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. in questing Ixodes ricinus ticks.     

Prevalence of Babesia canis, Borrelia afzelii, and Anaplasma phagocytophilum infection in hard ticks removed from dogs in Warsaw (central Poland)

The purposes of this study were to specify the occurrence and prevalence of Babesia canis, Borrelia burgdorferi sensu lato, and Anaplasma phagocytophilum in ticks removed from dogs in Warsaw, and to determine the Borrelia species occurring in Ixodes ricinus ticks. Among 590 collected ticks, 209 were identified as I. ricinus, and 381 as Dermacentor reticulatus. DNA of B. canis was detected in 11% of D. reticulatus ticks. We found that 6.2% of I. ricinus ticks harbored B. burgdorferi s.l. specific DNA and 2.9% harbored A. phagocytophilum DNA. In these samples sequencing of the detected Borrelia amplicon confirmed infection with Borrelia afzelii genospecies. New sequences were submitted to the GenBank database (accession no. EU152128, EU152127, EU152126). This work is the first detection of B. afzelii and A. phagocytophilum in ticks from Warsaw, and the first survey for the prevalence of B. canis, B. afzelii, and A. phagocytophilum in ticks in central Poland.     

Liver abscesses in intensively fed cattle. Serial investigations of serum proteins

An attempt was made to determine the time of onset of liver abscesses in intensively fed calves. Determinations of seram proteins (total protein, albumin, globulin, albumin/ globulin ratio, agarose-gel electrophoretic separation and formol-gel test) and, in a part of the material, of serum-GOT and -GPT were made in serial samples taken once a month from 125 calves. Liver abscesses were found in 48 (38.4 %) when slaughtered at 6–7 months of age.Total protein and globulin increased, the albumin-globulin ratio decreased with increasing age. Calves with liver abscesses had a significantly (0.05 > P > 0.01) higher globulin concentration at the last sampling. At this sampling the material was of a limited size and the difference has to be interpreted with caution. No other significant differences were found. Nor were any rises observed in S-GOT and S-GPT referable to the formation of liver abscesses.Bacteriological studies were not made but, in analogy with observations on a similar animal material in another part of Sweden, the infectious agent in the liver abscesses was assumed to be Sph. necrophorus. The explanation of the almost absent changes in the serum proteins in calves with liver abscesses may be the low antigenicity of this bacterium.In conclusion it is stated that the present serial investigations of the serum protein pattern in young cattle provide no definite guidance for establishing the time of onset of liver abscesses.     

Detection of bovine antibodies to the outer membrane of ruminal Bacteroides succinogenes by enzyme-linked immunosorbent assay (ELISA)

The enzyme-linked immunosorbent assay (ELISA) was used to detect bovine serum antibodies directed to the outer membrane antigen of a ruminal bacteria, Bacteroides succinogenes. The outer membrane antigen of B. succinogenes was highly reactive against homologous antiserum, compared with rabbit sera raised against B. ruminicola subsp. ruminicola, B. ruminicola subsp. brevis and Selenomonas ruminantium. The titers of sera from colostrum-deprived calves were negligible level, while those of sera from colostrum-fed calves were relatively high. The mean titer of sera from 10 day-old calves was significantly (p less than 0.01) higher than that of 40 day-old calves, and was significantly (p less than 0.01) lower than that of adult cattle. The mean titer of sera from dairy cows which fed high-roughage diet was higher than that of feedlot cattle which fed high-concentrate diet. These results suggest that the antibodies against the outer membrane antigen of B. succinogenes transfer to calves via the colostrum, and that the titers of cows are affected by the way of feed management.     

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.     

Epidemiological studies of the infection of ticks with borreliosis agents in small mammals from north Germany

In a two years study into the infestation of ticks with Borrelia burgdorferi from mice in North Germany 1330 mice out of 11 species could be examined. Altogether 508 mice showed to be parasitized by 1445 ticks belonging to three species of Ixodes. 777 I. ricinus from 334 mice could be tested for B. burgdorferi. In 66 ticks (8.5%) from 34 mice (10.2%) borreliae could be demonstrated. These discoveries came from 9 of 14 investigated forest regions in Lower Saxony.     

Arthropoda as zoonoses and their implications.

Some aspects of changing patterns of arthropodal infections and arthropod-borne diseases in Mediterranean areas are briefly discussed. Selected examples are given, with particular emphasis on the phenomenon of the synanthropic flea Ctenocephalides felis felis and on health problems caused by human infections with Argas reflexus, the common tick of urban pigeons in Europe. Finally, the risk of the emergence of Lyme borreliosis (Borrelia burgdorferi) is considered in relation to the increasing spread of environmental infestation with ticks, mainly Ixodes ricinus, an efficacious vector for the spirochaete.     

Leptospirosis in beef herds from western Canada: serum antibody titers and vaccination practices

One study described the frequency of pre-breeding vaccination for leptospirosis in 205 cow-calf herds from across western Canada and the prevalence of positive Leptospira antibody titers in unvaccinated, weaned calves from 61 of these herds. The percentages of herds vaccinated for leptospirosis were 13.7% in 2001 and 8.4% in 2002. Of 1539 calves examined, 13 (0.8%) had a positive antibody titer for a Leptospira serovar; the most common serovar detected was hardjo. A second study examined the prevalence of positive Leptospira antibody titers during the summer grazing season in 313 vaccinated and 478 unvaccinated cows from 40 cow-calf herds in southern Saskatchewan. Antibody titers for 7 Leptospira serovars were measured during the grazing season. Of the non-vaccinated cows, 9.6% were positive in the spring for serovar pomona, 6.7% for serovar grippotyphosa, and 6.1% for serovar icterohaemorrhagiae; the corresponding percentages for the fall were 5.5%, 3.0%, and 1.3%, respectively. Of 781 vaccinated and unvaccinated cows that were sampled twice, 11.3% of vaccinated cows and 2.3% of unvaccinated cows had increases in Leptospira antibody titers during the grazing season.     

The selenium content of milk

The selenium content in milk of dairy cows from different countries varies between 2 and 60 micrograms/kg, because of differences in the selenium content in the feeds. With an increasing selenium content in the feed a decreasing part is secreted into the milk. A bigger part is utilized from natural selenium compounds in the feed than from selenite. In own investigations milk from Swedish cows had significantly lower selenium content during summer and autumn than during winter and spring (p less than 0,0001). The LS-means between different regions also differed significantly (p less than 0,0001). From the autumn of 1980 the feed manufacturers have been allowed to add sodium-selenite to concentrates and mineral feeds in amounts permitting a selenium content of 0,1 mg/kg DM in the total ration of dairy cows. The year after the selenium fortification was allowed, the LS-means for the selenium content of milk were 1 microgram/kg higher than the year before (p less than 0,001). In the southern parts of Sweden the selenium content was 8-10 and in the central and northern parts 7-9 micrograms/kg. These means were substantially lower than the figure of 15 micrograms/kg which was reported from the central part of Sweden two decades ago. The low selenium content in the milk during summer in some parts of Sweden could possibly mean that there is still a risk of selenium deficiency among the cattle. Anyhow the milk will not always cover the selenium requirement of the sucking or milkfed calf.     

A disease resembling louping-ill in sheep in the Basque region of Spain

A clinical syndrome resembling louping-ill which primarily affects lambs and yearlings in the Basque region of Spain is described. The disease has been observed for several years during May and June after the sheep flocks have been taken to the mountain grazings where the sheep tick (Ixodes ricinus) is known to occur. Examination of the brain from one of the affected animals revealed histological changes indistinguishable from those caused by louping-ill virus. In addition antibody that reacted with louping-ill virus antigen was detected in the serum of 57 per cent of the sheep tested from the affected flocks but in only 0.8 per cent of sera from flocks free of the disease. These preliminary results suggest that louping-ill or a related tick-borne virus is responsible for ovine encephalomyelitis in Spain.     

全株玉米青贮、谷草和羊草组合全混合日粮饲喂干奶前期奶牛对其围产期生产性能和血液生化及免疫指标的影响

在研究不同比例全株玉米青贮、谷草和羊草组合日粮饲喂干奶前期奶牛对其围产期生产性能和血液生化及免疫指标的影响。选择健康、体况一致的干奶前期荷斯坦奶牛45头,根据体重、胎次及预产期接近(P>0.05)的原则将奶牛随机分为试验A、B和C组,每组15头。分别以精料、全株玉米青贮、谷草和羊草16∶24∶30∶30(A组)、16∶24∶45∶15(B组)和16∶24∶60∶0(C组)比例配合日粮饲喂。产前2周将3组试验牛转入围产牛群,3个试验组开始饲喂场内同一围产前期和新产牛日粮。试验期88 d。结果表明:1)与C组相比,B组在产后1、2和3周的干物质采食量分别提高10.46%(P<0.05)、7.79%(P<0.05)和5.65%(P<0.05)。除产后1周外,产后2、3周B组的奶牛体况评分分别比C组高6.60%(P<0.05)和9.06%(P<0.05)。2)各试验组的犊牛初生重差异不显著(P>0.05)。试验A、B组初乳免疫球蛋白G(IgG)含量分别比C组提高10.82%(P<0.01)和18.07%(P<0.01)。3)与C组相比,B组产后21 d产奶量提高9.00%(P<0.05),乳非脂固形物率提高2.83%(P<0.05),乳蛋白率提高10.59%(P<0.05),乳糖率提高5.64%(P<0.05)。各试验组间乳脂率和尿素氮无显著差异(P>0.05)。4)分娩当天和产后21 d,B组的血清甘油三酯含量显著低于A和C组(P<0.05),B组的血糖含量显著高于A和C组(P<0.05), B组的血清尿素氮和胰岛素显著低于C组(P<0.05),B组的胰岛素样生长因子I(IGF-I)和总蛋白显著高于C组(P<0.05);在产前7 d和分娩当天,B组的血清免疫球蛋白A(IgA)、免疫球蛋白B(IgM)和IgG显著高于C组(P<0.05);在产后21 d,B组的血清IgM和IgG含量显著高于C组(P<0.05);产前7 d和分娩当天,B组瘦素显著高于C组,而B组的血清非酯化脂肪酸(NEFA)含量显著低于C组(P<0.05);在产前7 d、分娩当天和产后21 d,B组的胆固醇均显著低于C组(P<0.05),B组的血清白蛋白含量显著高于C组,B组谷草转氨酶(AST)和谷丙转氨酶(ALT)也显著低于C组(P<0.05)。各试验组间β-羟丁酸无显著差异(P>0.05)。综合分析,干奶前期饲粮组成和养分水平可影响围产期奶牛的机体代谢和生产性能。在本试验条件下,干奶前期奶牛日粮中精料、全株玉米青贮、谷草和羊草的适宜比例为16∶24∶45∶15。     

Evaluation of different media and a BGM cell culture assay for isolation of Borrelia burgdorferi sensu lato from ticks and dogs

A co-culture assay for isolation of Borrelia burgdorferi sensu lato (sl.) from naturally infected ticks and dogs suspected of Lyme borreliosis (LB) was evaluated using buffalo-green-monkey (BGM) cells as the mammalian component. Four different media were tested for their ability to provide sufficient growth conditions for spirochetes and BGM cells. A total of 176 Ixodes ricinus ticks and 268 specimens from 98 dogs were used to compare cell-free culture with the BGM co-culture. A 1:1 mixture of Barbour-Stoenner-Kelly medium (BSK) and Eagle's minimum essential medium (EMEM) supported the growth of the two test strains, B. burgdorferi sensu stricto B31 and B. valaisiana VS116 to the same extent as BSK medium and the growth as well as the viability of BGM cells in this medium were the same as in EMEM. Using the 1:1 mixture of BSK and EMEM, borrelial growth measured in co-culture with BGM cells did not differ significantly from corresponding values obtained in cell-free cultures. In cell-free culture the isolation rate of B. burgdorferi sl. from ticks was significantly higher in BSK/EMEM 1:1 than in BSK medium (P < 0.01). Co-culture with BGM cells had no significant influence on the isolation rate of borreliae from ticks. However, a significant amount of isolates were obtained by one of the procedures only. Analysing canine specimens accordingly, spirochetes were grown from the blood of one dog after four weeks in BGM cell co-culture. The isolate was classified as B. afzelii by PCR-coupled restriction fragment length polymorphism analysis.     

Experimental infection of the white-footed mouse with Borrelia burgdorferi

Laboratory raised white-footed mice (Peromyscus leucopus) were inoculated experimentally with live spirochetes (Borrelia burgdorferi), the etiologic agent of Lyme disease (borreliosis). Prior to inoculation, mouse sera were tested with an indirect fluorescent antibody test, and all mice were seronegative. All inoculated mice seroconverted. In tick transmission studies, immature stages of Ixodes dammini and Dermacentor variabilis attached and fed to repletion on mice, but only I dammini transferred spirochetes to uninfected mice. Mice were susceptible to oral infection and transmitted infection to each other through direct contact. Infection did not affect reproduction or development of young born from infected dams, nor did spirochetes appear in the tissues of neonates. Borrelia burgdorferi spirochetes were identified in the kidneys, liver, and spleen of infected mice by the use of the modified Steiner silver stain and a tissue indirect fluorescent antibody test. Spirochetes also were isolated in culture of the heart blood of 1 mouse. Regardless of the source of infection, no mice developed clinical signs or had any pathologic change resulting from infection. Spirochetes were always observed extracellularly within interstitial spaces. Antibody titers persisted for over 4 months in some mice and spirochetes were found in the tissues of 1 mouse that had been infected 1 year earlier.     

Rheumatoid arthritis subsequent to Borrelia burgdorferi infection in two dogs

Two dogs with clinical signs of polyarthritis developed rheumatoid arthritis subsequent to Borrelia burgdorferi infection. In both dogs, the diagnosis of B burgdorferi infection was based on clinical signs of disease and high serum B burgdorferi titer. After antibiotic administration, both dogs had decreased B burgdorferi titer, but clinical response was temporary or was lacking. The dogs subsequently were rheumatoid factor-positive (antinuclear antibody- and anti-globulin-negative) and responded to anti-inflammatory drug administration. Development of rheumatoid arthritis in both dogs after B burgdorferi infection implicates the Borrelia organism as an infective agent leading to the development of rheumatoid arthritis in dogs. Dogs with clinical signs suggestive of B burgdorferi infection should have antiglobulin, anti-nuclear antibody rheumatoid factor, and B burgdorferi tests performed to aid definitive diagnosis.     

Morbidity in Swedish dairy calves from birth to 90 days of age and individual calf-level risk factors for infectious diseases

The health of 3081 heifer calves born in 122 dairy herds in the south-west of Sweden from 1 January to 31 December, 1998, was monitored from birth until 90 days of age. The calves were kept either in individual pens (n=2167), in group pens, with 3–8 calves to a pen and manual feeding of milk (n=440), in group pens with 6–30 calves per pen and an automatic milk-feeding system (n=431), or with their dams (n=43). Disease incidence was recorded by farmers and project veterinarians, who clinically examined the calves and auscultated their lungs every 2–3 months. A disease was graded as ‘severe’ if the general loss of condition or of appetite in the calf continued for >2 days or if the animal suffered severe weight loss due to the disease. The effects of season, breed, housing, and type of colostrum feeding, and time, place and supervision of calving on the incidences of diarrhea, severe diarrhea, respiratory disease, other infectious disease and moderately to severely increased respiratory sounds, were analyzed by logistic-regression models (with herd as a random effect). The total morbidity rate was 0.081 cases per calf-month at risk. Incidence rates of arthritis, diarrhea, omphalophlebitis, respiratory disease and ringworm were 0.002, 0.035, 0.005, 0.025 and 0.009 cases per calf-months at risk, respectively. The odds ratios for diarrhea and severe diarrhea were increased in Swedish Red and Whites (OR: 1.6, 2.3) and in calves that received colostrum from first-lactation cows (OR: 1.3–1.8), and for severe diarrhea in calves born in summer or that received colostrum through suckling (OR: 1.7, 1.8). The odds ratios for respiratory disease and increased respiratory sounds were increased in calves housed in large-group pens with an automatic milk-feeding system (OR: 2.2, 2.8). Supervision of calving was associated with a decreased odds ratio for respiratory disease (OR: 0.7) and birth in individual maternity pen or tie stalls with a decreased odds ratio for increased respiratory sounds (OR: 0.5–0.6). Cross-breeds with beef breeds were associated with increased odds ratios for increased respiratory sounds (OR: 2.1–4.3) and colostrum from second-lactation cows and birth during night for other infectious disease (OR: 1.6, 1.5).