Herd-level Coxiella burnetii seroprevalence was not associated with herd-level breeding performance in Czech dairy herds

Q fever was studied on a large agricultural farm in northern Moravia, Czech Republic. Antibodies to Coxiella burnetii were ascertained by a complement fixation test. Titre of 8 or higher was considered as positive. The seroprevalences in cows (each cow was examined only once immediately after drying-off during a one-year period) from 14 different herds was between 4 to 19%. No significant correlation between seroprevalence levels and fertility characteristics in the cow herds was found. Of a total of 17 samples of milk from 17 randomly-selected cows with antibody titres of 8 to 32, one strain of Coxiella burnetii was isolated. In heifers (n = 339) and beef bulls (n = 210) no antibodies to C. burnetii were found. In cattle from the farm investigated, latent Q fever did not represent any health problem.     

Multiplication of Staphylococci in vitro in normal and mastitic milk from vaccinated and nonvaccinated cows

Mastitis was induced by injection of cell walls of Staphylococcus aureus into the mammary glands of normal cows and of cows which had been vaccinated parenterally with a staphylococcal bacterin in adjuvant. Multiplication of S aureus in normal milk and in mastitic milk from vaccinated and nonvaccinated cows was determined in constant volume cultures. Growth was significantly inhibited during the 1st 6 hours of incubation, regardless of the nature of the milk or the vaccination status of the cows. Growth was inhibited for 24 hours in normal milk, and the organisms grew exponentially in mastitic milk regardless of the vaccination status of the cows.     

Simultaneous transfer of free fibula and radial forearm flaps for complex oromandibular reconstruction

In the present investigation Actinomyces pyogenes specific antigens caused an antibody response in the host. This could be determined by two enzyme-linked immunosorbent assays (ELISA) using cell extracts and a haemolysin preparation as antigens. An increased antibody titre was detectable in the sera of cows vaccinated with culture supernatants of A. pyogenes and Peptococcus indolicus, in the sera of cows infected with live cells of A. pyogenes and P. indolicus, in the sera of cows suffering from 'summer mastitis', and in part of the sera of nonvaccinated, apparently healthy cows. However, the titre of the cows that were vaccinated with culture supernatants decreased 8-9 months after inoculation. Because of the wide variation in antibody titre the determination of A. pyogenes specific antibodies seems to be only of limited use for the control of this infection.     

Effects of vaccination with a Moraxella bovis bacterin on the subsequent development of signs of corneal disease and infection with M bovis in calves under natural environmental conditions

A vaccination study was conducted for infectious bovine keratoconjunctivitis (IBK) in 440 purebred Hereford cattle (cows and their newborn calves) of the USDA Meat Animal Research Center cattle herd at Clay Center, Ne. The cattle were allotted to 4 groups: 60 calves were vaccinated with an autogenous Moraxella bovis bacterin (group 1); 60 calves that were matched with group 1 calves were designated nonvaccinated matched controls (group 2); 99 calves were peer group nonvaccinated controls (group 3); and 219 cows, the dams of the calves, were nonvaccinated consorts (group 4). The infection rates in cattle groups 1, 2, 3, and 4 during the summer were 96.6, 98.3, 100, and 79.1%, respectively, and the disease rates were 90, 93, 85, and 20%. The infection and the disease rates were significantly (P less than 0.01) different between claves and cows. The disease rate was also significantly different between older and younger cows. A larger percentage of the affected calves and cows had mild or moderate (61%) signs of IBK rather than severe (39%) signs. The rate of body weight gain was reduced in calves with severe signs of IBK. The results seemed to indicate that little would be gained by vaccinating cattle against IBK under the conditions of study.     

The use of atomic-force microscopy for the analysis of microbiological objects

Atomic force microscopy (AFM) was used for the analysis of rickettsiae and viruses. The specificity of interaction was evaluated on the basis of the adsorption of the analyzed antigen on the polymer-antibody film. The film was formed and transferred onto highly oriented pyrolytic graphite (HOPG) by the method of Langmuir-Schaefer with the use of amphiphilic polymers, alkylated polyethyleneimines. According to the data of AFM, polymer-antibody film was 10-30 nm thick and had ruptures, uneven surface. AFM images of Coxiella burnetii, rotavirus and Venezuelan equine encephalitis virus, immunoadsorbed on the antibody film, were obtained. C.burnetii, due to their size equal to (700-900) x (300-500) x Z = 200 nm, were clearly visible on the underlying surface and could be directly counted. Individual virus particles (60-80 nm) cold not be identified on such surface. To analyze such preparations, the program of image analysis was developed. The program classified the registered image with a certain standard. This program determined the presence of virus antigen on the underlying affinity surface with a high degree of precision.     

Investigation of film curing stages by dielectric analysis and physical characterization

A previously published sequence of the 23S rRNA gene of Coxiella burnetii has been reported to contain an intervening sequence of 444 base pairs (bp). The sequence information on the intervening sequence and the 23S rRNA gene was exploited to develop a specific PCR-based assay for C. burnetii. A primer set was designed that amplified a 477-bp fragment encompassing part of the intervening sequence and part of the 23S rDNA. From all of nine C. burnetii strains tested, a fragment of the expected size was amplified. As predicted from the published sequence, restriction endonuclease digestion of the PCR product from the Coxiella strains with RsaI produced two distinct fragments approximately 210- and 270-bp in size. The PCR-based method showed a detection limit of 10(2) bacteria as determined by visualization of the amplicon on an agarose gel. When experimentally infected blood was analyzed, the detection limit was 10(3) bacteria. No visible amplicons were observed when 41 bacterial strains, representing 29 species other than C. burnetii, were tested. The presence of the DNA in all bacterial samples was confirmed by amplification of a 350-bp fragment of the 16S rDNA using two universal primers. The described method proved to be specific for C. burnetii and may become a rapid and sensitive diagnostic assay for C. burnetii. The results also demonstrate that the intervening sequence within the 23S rRNA gene is generally found among isolates of C. burnetii.     

Fever, negative blood culture findings and absence of response to antibiotic therapy in a patient after a second aortic valve prosthesis

HISTORY AND CLINICAL FINDINGS: A 53-year-old patient had a prosthetic valve (St. Jude Medical 25) 9 years ago because of a Staphylococcus aureus endocarditis with severe aortic regurgitation. An initially mild, progressively more severe, aortic regurgitation then developed as a result of an empty paravalvular abscess cavity, requiring another valve replacement. Fever started on the 3rd postoperative day and persisted despite combined treatment with beta-lactam antibiotics and aminoglycoside. INVESTIGATIONS: At first no infectious focus could be identified radiologically or by echocardiography. But transoesophageal echocardiography revealed vegetations in the old abscess cavity. Several blood cultures were negative, while serological tests gave markedly raised antibody titers against Coxiella burnetii. DIAGNOSIS, TREATMENT AND COURSE: Assuming Coxiella burnetii endocarditis the patient was given doxycycline, 2 x 100 mg daily and cotrimoxazole, 1 x 960 mg daily. The fever subsided and the vegetations had disappeared after four weeks. Because of the high risk of recurrence the antibiotic treatment was to be continued for two years. CONCLUSION: Coxiella burnetii should be considered as a possible cause of fever of unknown origin, especially in patients with existing or operated cardiac valvar defects, when endocarditic vegetations have been demonstrated and several blood cultures have been negative.     

Frequency and cost of health problems in Swiss dairy cows and their calves (1993-1994)

Between July 1993 and July 1994 morbidity and management information related to dairy cows and their calves up to the age of 8 weeks were recorded in 113 randomly selected dairy herds. Also recorded were any costs incurred through disease and prevention. Blood and faeces were analysed with respect to selected pathogens. The health problems most frequently diagnosed in cows were reproductive and udder diseases. Calves suffered most often from diarrhea, omphalitis and pneumonia. The directly disease-related costs per cow-year on average amounted to CHF 139.44 and CHF 4.18 per calf. For prevention, farmers spent on average CHF 10.18 per cow-year. Results from the laboratory analyses indicate that in 68.1% of the farms antibodies against Leptospira hardjo and in 61.9% against Coxiella burnetii were detected. In 8.0% of the farms antibodies against Mycobacterium paratuberculosis were found. Antibodies against BVD virus was present in 99.4% of the farms. Cows from 63.7% farms were infected with gastrointestinal strongylids. Veterinary assistance was required on average 1.96 times per cow-year. In almost all reproductive and puerperal disease cases a veterinarian was consulted while lameness in the majority of cases was treated by the owner. The veterinary profession was hardly ever involved in disease prevention.     

Phaco-emulsification of cataract in eyes with glaucoma

Previous observations on the dissemination of Coxiella burnetii between laboratory animals strongly support the hypothesis of venereal transmission. Serum and semen samples, from 57 bulls used for artificial insemination, were assayed for specific C. burnetii phase II antibodies and the presence of the organism respectively. Viable C. burnetii were detected in the semen of seropositive bulls. These findings indicate the possibility of sexual transmission of C. burnetii between cattle and further our knowledge of the epidemiology of the organism. The procedures used for investigations into the source of infection and route of tran-mission should be modified to take these findings into account.     

IFN-gamma-mediated control of Coxiella burnetii survival in monocytes: the role of cell apoptosis and TNF

The treatment of infectious diseases caused by intracellular bacteria, such as Q fever, may benefit from cytokines acting on macrophages. Monocytic THP-1 cells were infected with Coxiella burnetii, the etiological agent of Q fever, and then treated with IFN-gamma. While C. burnetii multiplied in untreated monocytes, IFN-gamma reduced bacterial viability after 24 h of treatment and reached maximum inhibition after 96 h. IFN-gamma also affected the viability of infected cells. Cell death resulted from apoptosis; occurring 24 h after the addition of IFN-gamma, it reached a maximum after 48 h and was followed by necrosis. Reactive oxygen intermediates were not required for C. burnetii killing, since monocytes from patients with chronic granulomatous disease were microbicidal in response to IFN-gamma. The role of cytokines was also investigated. IFN-gamma elicited a moderate release of IL-1beta in infected monocytes. Moreover, the IL-1 receptor antagonist did not affect C. burnetii survival, suggesting that IL-1beta was not involved in the bacterial killing induced by IFN-gamma. TNF was involved in IFN-gamma-induced killing of C. burnetii and cell death. IFN-gamma induced mRNA expression and sustained secretion of TNF. Neutralizing Abs to TNF as well as Abs directed against TNF receptors I and II, significantly prevented IFN-gamma-dependent killing of C. burnetii and cell death. These results suggest that IFN-gamma promotes the killing of C. burnetii in monocytes through an apoptotic mechanism mediated in part by TNF.     

Efficacy of a live avirulent Salmonella typhimurium vaccine in preventing colonization and invasion of laying hens by Salmonella typhimurium and Salmonella enteritidis

An avirulent live delta cya delta crp Salmonella typhimurium strain chi 3985 that precludes colonization and invasion of chickens by homologous and heterologous Salmonella serotypes was evaluated for its long-term protection efficacy. Chickens vaccinated orally at 2 and 4 wk of age were assessed for protection against oral challenge with wild-type S. typhimurium and Salmonella enteritidis strains at 3, 6, 9, and 12 mo of age. A comparison of Salmonella isolation from vaccinated and nonvaccinated layers after challenge with S. typhimurium or S. enteritidis showed that delta cya delta crp S. typhimurium chi 3985 induced excellent protection against intestinal, visceral, reproductive tract, and egg colonization, invasion, and/or contamination by Salmonella. The duration of protection lasted for 11 mo after vaccination, at which time the experiment was terminated. S. enteritidis and S. typhimurium were isolated from the yolk, albumen, and shells of eggs laid by nonvaccinated chickens challenged with Salmonella. S. typhimurium caused pathological lesions in nonvaccinated chickens, whereas vaccinated and nonvaccinated chickens challenged with S. enteritidis showed no pathological lesion in the visceral and reproductive organs. Vaccination with chi 3985 prevented transmission of S. typhimurium or S. enteritidis into eggs laid by vaccinated layers with no effect on egg production. To our knowledge, this is the first publication confirming that vaccination with live avirulent Salmonella can induce long-term protection against Salmonella infection in layers.     

Isolation of Coxiella burnetii from dairy cattle and ticks, and some characteristics of the isolates in Japan

Coxiella burnetii was isolated from raw milk (36/214, 16.8%) and uterus swab samples (13/61, 21.3%) originating from dairy cattle with reproductive disorders, aborted bovine fetus samples (2/4, 50%), mammary gland samples (4/50, 8%) originating from healthy dairy cattle, and tick samples (4/15, 26.7%) originating from 2 pastures. Fifty-nine strains had various degrees of pathogenicity, high (8; 13.6%), moderate (28; 47.5%) and low (23; 39%), for guinea pigs. The results of isolation suggested a high prevalence of Coxiella infection in dairy cattle with reproductive problems in Japan. Twelve strains (7, 2 and 3 strains from cattle, ticks and humans, respectively) and the reference Nine Mile strain of phases I and II were propagated in both yolk sacs of embryonated hen eggs and Buffalo green monkey (BGM) cell cultures. Protein profiles of these strains were similar to those of the reference strain of phase I. Lipopolysaccharide (LPS) profiles of 12 strains were similar to those of the reference strain of phase I and different from those of the reference strain of phase II. The LPS profiles of 12 strains suggested that these strains are associated with an acute form of Q fever.     

Prevalence of Coxiella burnetii infection in dairy cattle with reproductive disorders

The prevalence of Coxiella burnetii infection in 207 cattle with reproductive disorders was studied by using an indirect immunofluorescence (IF) test, nested polymerase chain reaction (PCR) and isolation. IF antibodies to phase I and phase II antigens of C. burnetii were found in 122 (58.9%) and 125 (60.4%) of the sera, respectively, and PCR-positives were found in 8 (3.9%) of the sera and in 51 (24.6%) of the milk samples. In addition, C. burnetii was isolated from 51 (24.6%) of the milk samples by inoculating laboratory mice. The results indicate that the IF test plus PCR are useful in the diagnosis of bovine coxiellosis. It is difficult to deny that dairy cattle with reproductive disorders would be one of the important reservoirs of C. burnetii responsible for infection in both animal and human populations in Japan.     

In vitro susceptibility of Coxiella burnetii to trovafloxacin in comparison with susceptibilities to pefloxacin, ciprofloxacin, ofloxacin, doxycycline, and clarithromycin

The antibiotic susceptibilities of eight Greek isolates of Coxiella burnetii to trovafloxacin were determined by the shell vial assay. MICs of trovafloxacin and ofloxacin ranged from 1 to 2 microg/ml, those of pefloxacin ranged from 1 to 4 microg/ml, those of ciprofloxacin ranged from 4 to 8 microg/ml, those of doxycycline ranged from 1 to 2 microg/ml, and those of clarithromycin ranged from 2 to 4 microg/ml. Trovafloxacin exhibited no activity against C. burnetii at 4 microg/ml.     

Coxiella burnetii penetration into the reproductive system of male mice, promoting sexual transmission of infection

Coxiella burnetii bacteria penetrate different host organs via the bloodstream. In infected mice, bacteremia was observed during the first week of infection: later, bacteria were cultured from the spleens, livers, testes, epididymes, prostate, and semen; bacteriuria developed beginning from the second week of infection. On day 21 of infection, degenerative changes with aggregated macrophages containing bacteria were observed in capillary blood vessels and the surrounding tissues of the adipose envelope of the epididymis. C. burnetii was shed to semen from the urogenital tract, probably from an abscess in the epididymis. There they were bound to the surface of spermatozoal cells, mainly to their heads, suggesting specific adhesion. Bacteria shed to semen were transmitted to female mice by sexual contact; this was demonstrated by the detection of antibodies against C. burnetii antigens in the blood of females and later by the cultivation of bacteria from the spleens, livers, and amniotic fluids of female mice.     

Attitudes of oncologists, family doctors, medical students and lawyers to euthanasia

OBJECTIVE: To evaluate clearance of the vaccine strain, immunologic responses, and potential shedding of Brucella abortus strain RB51 organisms after vaccination of bison calves. ANIMALS: Fourteen 7-month-old female bison calves. PROCEDURE: 10 bison calves were vaccinated SC with 1.22 x 10(10) colony-forming units of B abortus strain RB51. Four bison calves were vaccinated SC with 0.15M NaCl solution. Rectal, vaginal, nasal, and ocular swab specimens were obtained to evaluate potential shedding by vaccinated bison. The superficial cervical lymph node was biopsied to evaluate clearance of the vaccine strain. Lymphocyte proliferative responses to strain RB51 bacteria were evaluated in lymph node cells obtained from biopsy specimens and also in peripheral blood mononuclear cells. RESULTS: Strain RB51 was recovered from superficial cervical lymph nodes of vaccinates examined 6, 12, and 18 weeks after vaccination (4/4, 3/4, and 1/4, respectively) but not in vaccinates examined at 24 weeks (0/3) after vaccination or nonvaccinates examined at all sample collection times (n = 1 bison/sample period). Serologic, immunologic, and bacterial culture techniques failed to reveal shedding of strain RB51 by vaccinates or infection of nonvaccinated bison. Lymphocyte proliferative responses were evident in lymph node cells and blood mononuclear cells from strain RB51-vaccinated bison beginning 12 weeks after vaccination. CONCLUSION: Strain RB51 was cleared from bison by 18 to 24 weeks after vaccination. Bison vaccinated with strain RB51 did not shed the vaccine strain to nonvaccinated bison housed in close proximity. Strain RB51 did not induce antibody responses in bison that would interfere with brucellosis surveillance tests, but did stimulate cell-mediated immunity.     

Vaccination against feline pneumonitis

A commercially available modified live chlamydial vaccine against feline pneumonitis was tested in 26 cats for its ability to protect against aerosol challenge exposure to the feline pneumonitis strain of Chlamydia psittaci. After cats were challenge exposed (30 days after vaccination), pyrexia of greater than 40.0 C occurred in 81% of nonvaccinated (control) cats and in 13% of vaccinated cats (principals). Evidence of upper respiratory tract disease and the presence of the agent in ocular fluids were observed less frequently in principals than in nonvaccinated cats. In the cats euthanatized at intervals of 3 days after challenge exposure, C psittaci was demonstrated in 60% of tissues tested from nonvaccinated controls and in 34% of similar tissues obtained from principals.     

Studies of the prevalence of Coxiella burnetii, the agent of Q fever, in the foothills of the southern Bavarian Forest, Germany

In studies carried out in 1991 in the foothills of the southern part of the Bavarian Forest, in the district of Freyung/Grafenau, ticks and small mammals were collected and examined for the presence of Coxiella (C.) burnetii and sera of small mammals and cattle investigated for antibodies against this rickettsia. A total of 1716 imagines and nymphs of Ixodes ricinus were collected by flagging and 892 larvae and nymphs of the same tick species removed from small mammals. In addition to 1095 serum samples from cattle, 326 specimens of nine species of small terrestrian mammals were examined. Neither in ticks nor in rodents, C. burnetii was detected, however, in 17 of 21 localities, seropositive cattle were found. Altogether, 12% of all 1095 heads of cattle tested were seropositive for C. burnetii antibodies. These serological results indicated a wide dissemination of C. burnetii in cattle of the region investigated, but there was no indication of a natural focus. As in other areas of Europe, an independent natural cycle of the agent involving cattle only is assumed to occur in this region.     

Anti-C1q receptor/calreticulin autoantibodies in patients with systemic lupus erythematosus (SLE)

We report a case of acute glomerulonephritis associated with acute Q fever. An abattoir worker with a nonspecific febrile illness and pneumonia and abnormal liver function test results developed hematuria, proteinuria, and acute renal failure that resolved with appropriate antimicrobial therapy. Renal biopsy demonstrated diffuse proliferative and exudative glomerulonephritis. Serological tests confirmed recent infection with Coxiella burnetii, with a fourfold rise in the titer of phase II antibody, positive phase II IgM antibody, and negative phase I antibody. Other known causes of glomerulonephritis were excluded. Most reports of renal complications of C. burnetii infection describe glomerulonephritis associated with endocarditis due to chronic Q fever. Renal involvement in patients with acute C. burnetii infection has been rarely described. Glomerulonephritis should be recognized as a complication of acute C. burnetii infection and endocarditis due to chronic Q fever.     

The insulin receptor substrate 1 associates with phosphotyrosine phosphatase SHPTP2 in liver and muscle of rats

OBJECTIVE: To determine efficacy of a vaccine containing modified-live bovine viral diarrhea virus (BVDV) type 1 for protecting pregnant cows and their fetuses against virulent heterologous BVDV type 1. DESIGN: Randomized controlled cohort study. ANIMALS: 18 yearling beef heifers seronegative for BVDV and negative when tested for BVDV by virus isolation. PROCEDURE: Cattle were randomly assigned to control (unvaccinated; n = 6) or vaccinated (12) groups. Vaccinated heifers were given a combination vaccine containing modified-live BVDV type 1 comprising a cytopathic (NADL) strain. All 18 heifers were then bred and challenge-exposed between 70 and 75 days of gestation with BVDV type 1, administered intranasally. Cattle were monitored, and infection status of offspring was determined after parturition. Antibody concentrations of vaccinated and control heifers were also monitored. RESULTS: All 6 calves from control heifers had positive results on multiple virus isolation tests and were considered persistently infected. In comparison, only 2 calves from vaccinated cows had positive results on virus isolation tests and were considered persistently infected. One vaccinated heifer aborted, but the fetus was not persistently infected, and the abortion was not attributed to BVDV infection. CLINICAL IMPLICATIONS: Analysis of these data indicated that a single dose of a modified-live NADL-derived BVDV type 1 vaccine will confer protection to dams and their fetuses against challenge-exposure to heterologous BVDV type 1 organisms.