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1.
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.  相似文献   

2.
A micromethod was used in order to quantitate intestinal D-xylose absorption in young and extremely small birds. This test was performed in broilers collected from two farms from which birds were extremely uneven in body size and were passing poorly digested or undigested feed. A similar syndrome had been seen on all grow-outs during the 6 months before this investigation. Broilers were also collected and tested from three farms where no clinical signs of disease were seen. D-Xylose absorption peaks and curves for normal broilers closely resembled those observed in normal humans. Mean plasma D-xylose concentrations for virus-infected broilers and for broilers with pale-bird syndrome were consistently lower than concentrations for normal broilers (P = 0.009). Reoviruses, small coronavirus-like particles, small round virus particles, and abundant bacterial flagellar fragments were seen in fecal samples from broilers with pale-bird syndrome. Production performance was lowest on farms showing clinical signs of this syndrome.  相似文献   

3.
77 strains of Pasteurella from various animal species were tested using the API micromethod for their ability to assimilate 147 carbon substrates and ferment 49 carbohydrates. Typical profiles of P. multocida and P. haemolytica were determined. This micromethod should prove useful in establishing the biotypes of Pasteurella species and in studying their epidemiology and pathogenicity.  相似文献   

4.
According to the present study the limulus amebocyte lysate test (LAL) seems to be a convenient test to detect endotoxin in milk from udder quarters with and without inflammation. The correlation between endotoxin concentration and the results from the bacteriological investigation of 79 milk samples was good (Table I). Determination of endotoxin in 20 milk samples from cases of acute clinical mastitis with high cell count and a negative bacteriological culture showed that all but one had an endotoxin concentration of greater than 1.0 ng/ml milk (Table II). By using a micromethod of the LAL it is possible to detect cases of mastitis caused by gram-negative bacteria about one hour after the sample has reached the laboratory. In a preliminary field study milk from 13 cases of acute clinical mastitis were tested by a modified limulus amebocyte lysate (LAL) test ("cowshed test"). A 100% correlation to bacteriological findings was observed (Table IV). By using the LAL test to detect mastitis cases caused by gram-negative bacteria great economic advantages and less risk for resistance problems can be achieved by using proper antibiotics. This is the fact in Sweden where the frequency of acute clinical mastitis caused by streptococci (100% of strains sensitive for penicillin) and Staphylococcus aureus (about 90% of strains sensitive for penicillin) is high (70-80%) and about 20% are caused by gram-negative bacteria, mostly E. coli.  相似文献   

5.
A latex agglutination test was evaluated as a method for the detection and titration of antibodies against swine Haemophilus infection and it was found that the test is applicable to the etiologic diagnosis of Haemophilus infection in swine. In swine infected experimentally with H. pleuropneumoniae (H. parahaemolyticus), the micromethod of agglutination using latex particles coated with antigens of H. pleuropneumoniae was found to be comparable agar-gel immunodiffusion and complement-fixation tests, which have previously been used for the etiologic diagnosis of the disease. Antibody titers determined by the latex agglutination test corrlated well with those determined by the other serological tests. The latex agglutination test tended to detect antibodies earlier than any of the other tests. By the latex agglutination test, weak cross-reactions were observed among different serotypes of H. pleuropneumoniae, whereas no cross-reaction was demonstrated between H, pleurophneumoniae and H. parasuis. The latex agglutination test was found to be simple and useful for the serological survey of swine Haemophilus infection, especially when dealing with a large number of samples.  相似文献   

6.
A micromethod was modified to quantitate intestinal D-xylose absorption in young and extremely small birds. This test was performed in control and cloned-reovirus-inoculated turkey poults to investigate intestinal absorptive function. Absorption peaks and curves for control poults closely resembled those observed in normal humans. Poult groups absorbed significantly less D-xylose at 24 and 72 hours (P less than 0.0001 and P less than 0.0008, respectively) but not at 120 hours (P = 0.3178) after receiving a single oral dose of cloned reovirus. The modified D-xylose absorption test is suitable for experimental and clinical determination of absorptive function of the small intestine in the fowl.  相似文献   

7.
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8.
In order to rationalize work and to reduce the consumption of laboratory glassware a reduction micromethod with the bacterial stain Streptococcus thermophilus has been modified; the method is used to detect inhibitory substances in genuine, pausteurized and dried milk, and it is performed on a serological plate made from organic glass. The method is sensitive to most antibiotics used in the veterinary clinical practice. The detection sensitivity to penicillin residua ranges from 0.0015 to 0.0015 i.u. in 1 ml of milk. The method may be assessed as semi-quantitative.  相似文献   

9.
A simple and reproducible micromethod for determination of in vitro mitogenic responses of sheep peripheral blood lymphocytes is described. The test uses (i) whole blood diluted in RPMI 1640 medium to give a cell count of 0.5 x 106-1 x 106 lymphocytes/ml, (ii) mitogens in the range of 5-20 micrograms of phytohaemagglutinin/ml, 20-80 micrograms of lipopolysaccharide/ml or 20-80 microliters of poke weed mitogen/ml, and (iii) a stimulation time of 42 to 90 h. A considerable variation in mitogenic response was observed both between animals and on different occasions in the same animal. Because of the large periodic variation it was suggested that the test should be repeated using blood drawn at different times in order to determine the mitogenic response of an animal.  相似文献   

10.
The aim of this study was to determine the sensitivity of Aspergillus niger strains isolated from birds to available antifungal drugs using different in vitro assays--classical disk diffusion, Etest and broth microdilution NCCLS/CLSI M 38-A. The study material consisted of about 2.000 swabs and samples from different species of birds. A. niger (n=10) was accounted for 6.81% of the total pool of strains isolated. Determinations were made for 13 antifungal drugs using the disk diffusion method. The A. niger exhibited high susceptibility to enilconazole, terbinafine, voriconazole, tioconazole and ketoconazole, low susceptibility to clotrimazole, miconazole and nystatin, and resistance to amphotericin B, itraconazole, pimaricin, fluconazole and 5-fluorocytosine. Minimum inhibitory concentration (MIC) was determined for 9 antifungal drugs using the micromethod of duplicate serial dilutions in a liquid medium. A. niger strains were most susceptible to enilconazole and voriconazole. MIC ranged from 0.0625 to 0.5 microg/ml for enilconazole, with MIC90-0.5 microg/ml and MIC50-0.125 microg/ml. The corresponding values for voriconazole were 0.25-1 microg/ml, 1 microg/ml and 0.5 microg/ml. MIC for amphotericin B and terbinafine ranged from 0.5 to 4 microg/ml, while the values for the remaining drugs were highly varied. MIC was measured by the gradient diffusion method using Etest for 5 antifungal drugs: amphotericin B, fluconazole, itraconazole, ketoconazole and voriconazole. By far the highest susceptibility was obtained in the case of voriconazole, with MIC ranging from 0.0625 to 1 microg/ml. MIC for amphotericin B ranged from 0.25 to 4 microg/ml, for itraconazole and ketoconazole ranging from 0.5 to 16 microg/ml. Methods available for this purpose are not always applicable in field conditions. The present results indicate that the Etest technique, due to its high percentage of agreement with the M 38-A microdilution method, should find application in medical and veterinary practice.  相似文献   

11.
Fifty-one coagulase-positive staphylococcal isolates from canine urinary calculi or from the urine of dogs with documented urolithiasis, and 17 coagulase-positive staphylococcal isolates from human beings and cattle were identified by a commercially available tray micromethod, as well as by conventional methods. Canine isolates had previously been classified as Staphylococcus aureus on the basis of a positive tube coagulase test. After 5 hours' incubation, the tray method identified all 51 canine urolithiasis isolates as S intermedius, rather than S aureus. All human and bovine isolates were identified as S aureus. Conventional methods supported these findings.  相似文献   

12.
The in vitro sensitivity of 20 wild strains of M. capricolum and that of the reference strain (California kid) against 14 antibiotics was investigated by means of a micromethod. The technique is based on the determination of the inhibitory minimum concentration (IMC) in a liquid medium as revealed by the inhibition of glucose metabolism. The following results were obtained: all strains were sensitive to five antibiotics (tylosin, oxytetracyclin, gentamycin, neomycin, nalidixic acid) with an IMC varying from 0.06 to 8 meq/ml. The variation in the IMC values from 8 to 32 meq/ml for spiramycin and erythromycin indicated that some of these strains were sensitive and other resistant to these two drugs. All strains were resistant to seven antibiotics (streptomycin, bacitracin, polymyxin, chloramphenicol, lincomycine, rifampicine and novobiocine), sometimes at a concentration exceeding 128 meg/ml.  相似文献   

13.
A micromethod technique was used to evaluate in vitro sensitivity of the peripheric bovine lymphocytes obtained from a newly born calf, up to 3 months of age to different non-specific mitogens: Phytohemagglutinin (PHA) Concanavaline A (Con A and Pokeweed Mitogen (PWM). The results obtained show that the calf lymphocytes respond to the 3 mitogens by a considerable cellular proliferation. The blastogenic response was found at various levels during the first 3 months of life, and appeared to stabilize at levels similar to the adult bovine. Highly sensitive variations were noted in the lymphocyte reactivity, notably with PHA and Con A. These results seem to indicate the existence of periods of T cell immunodeficiency, not only during the first few days after birth, but throughout the first months of the calves' life. It may also be indicative of the interest of immunostimulant therapy during this period, which needs further investigation.  相似文献   

14.
Extracellular matrix components (ECMs) in histological sections of the kidney cortex from the rats with adriamycin (ADR)-induced nephropathy (5 mg/kg, i.v.) were quantified by an immunohistochemical micromethod. Changes in kidney histopathology and urine and blood biochemistry were investigated. Enlarged kidneys were granular on the surface and pale in color in ADR-treated rats, and these rats had kidneys with glomeruli with expanded mesangial area and with capillary aneurysm. Severe albuminuria, hypoalbuminemia, hypercholesterolemia and disorders in other nephrotic parameters were observed in ADR-treated rats. Type I and IV collagens, fibronectin and laminin contents in the renal cortex of ADR-treated rats at 10 weeks were 329, 317, 263 and 295%, respectively, higher than in each vehicle control, and those at 28 weeks were 1,211, 930, 1,057 and 1,012%, respectively. The glomerular sclerotic abnormalities progressed in a time-dependent manner. Moreover, there was a strong correlation between the ECM levels and serum creatinine and blood urea nitrogen levels. In conclusion, microquantification provided useful information for accurate diagnosis and prognosis of nephrotic lesions and is a good tool to assess the advancement of renal disorders in patients with nephropathy.  相似文献   

15.
Species of Staphylococcus isolated from animal infections   总被引：3，自引：0，他引：3  
One hundred randomly selected clinical strains of staphylococci were identified by species using a commercial micromethod system. Eight species of staphylococci were identified. Staphylococcus intermedius was the most frequent (n = 74) species identified and accounted for 70/74 (94.6%) of the coagulase-positive strains and 70/79 (88.6%) of the total isolates from dogs. Other species identified, in order of their frequency, included S. epidermidis (8), S. aureus (7), S. simulans (4), S. sciuri (2), S. xylosus (2), S. hyicus (2) and S. saprophyticus (1). These results show that at least 8 different species of staphylococci can be recovered from animal infections and that coagulase-positive species such as S. intermedius may be more common than S. aureus. The relative significance of these other species in animal infections needs to be assessed.  相似文献   

16.
Bovine coronavirus (BCoV) and group A bovine rotavirus (BRV) are two of major causes for neonatal calf diarrhea. In the present study, a one-step duplex RT-PCR was established to detect and differentiate BCoV and group A BRV from fecal samples. The sensitivity of this method for BCoV and group A BRV was 10 PFU/100 μl and 1 PFU/100 μl, respectively. Twenty-eight diarrhea fecal samples were detected with this method, the result showed that 2 samples were identified as co-infected with BCoV and group A BRV, 26 samples were group A BRV positive, and 2 samples were negative. It proved that this method is sensitive for clinical fecal samples and is worth applying to laboratory diagnosis for BCoV and group A BRV.  相似文献   

17.
The study describes a polymerase chain reaction (PCR) assay for the detection of Actinobacillus pleuropneumoniae. The test is based on the amplification of the omlA gene coding for an outer membrane protein of A. pleuropneumoniae. To test the specificity of the reaction, 19 other bacterial species related to A. pleuropneumoniae or isolated from pigs were assayed. They were all found negative in the PCR assay. The detection threshold of the test was 10(2) A. pleuropneumoniae CFU/assay. The test was then applied to the detection of A. pleuropneumoniae from tonsillar biopsies and tracheobronchial lavage fluids of pigs without a culture step. The detection of A. pleuropneumoniae in these samples was performed by PCR, by conventional culture and by bacteriology with immunomagnetic beads. The number of samples that were found positive by PCR was almost three times higher than the number of samples from which A. pleuropneumoniae was isolated by both bacteriological techniques. The detection of A. pleuropneumoniae in these samples allowed us to demonstrate its aerosol transmission to pigs under experimental conditions. The trial involved 18 specific pathogen free pigs. Six pigs, infected with A. pleuropneumoniae, were located in a unit A, together with four non-infected animals (contact pigs). Eight non-infected pigs (reporter pigs) were located in a unit B, adjacent to A. We detected A. pleuropneumoniae in samples from infected animals but also from 'contact' (unit A) and 'reporter' (unit B) pigs. The results of this study show that the simple preparation of the samples followed by the PCR assay may be a useful tool for epidemiological studies.  相似文献   

18.
A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) was described and evaluated for use as a presumptive screening test for detection of Campylobacter fetus in bovine preputial washing and vaginal mucus samples. A total of 725 diagnostic samples collected in the field and submitted in Clark's transport enrichment medium (TEM) were analyzed. Cultural isolation of C. fetus was used as the standard for comparison. After incubation of the TEM vials for 4-5 days, fluid was removed for culture and ELISA testing. A sandwich ELISA format was used and the target antigen was C. fetus lipopolysaccharides (LPS). A rabbit anti-C. fetus polyclonal antiserum was used as the capture antibody. Murine monoclonal antibodies (MAbs) to C. fetus serotype A and B LPS core and O-polysaccharides and a goat anti-mouse horseradish peroxidase conjugate were used as detection antibodies. ELISA and culture results for the diagnostic samples were in complete agreement. Seven hundred and eight samples were negative by both tests. All 17 culture positive samples were positive by ELISA with a MAb to LPS core. The ELISA with MAbs to LPS O-polysaccharides detected all culture positive samples with the homologous C. fetus serotype. Sixty-six preputial wash samples from three known C. fetus culture positive bulls were also analyzed. Forty-nine of these samples were positive by both ELISA and culture, 16 were positive by ELISA only, and one was negative by both ELISA and culture. The results indicate that this ELISA is useful as a screening test for the detection of C. fetus in diagnostic samples.  相似文献   

19.

Background

Swine influenza is an infectious acute respiratory disease of pigs caused by influenza A virus. We investigated the time of entry of swine influenza into the Finnish pig population. We also describe the molecular detection of two types of influenza A (H1N1) viruses in porcine samples submitted in 2009 and 2010.This retrospective study was based on three categories of samples: blood samples collected for disease monitoring from pigs at major slaughterhouses from 2007 to 2009; blood samples from pigs in farms with a special health status taken in 2008 and 2009; and diagnostic blood samples from pigs in farms with clinical signs of respiratory disease in 2008 and 2009. The blood samples were tested for influenza A antibodies with an antibody ELISA. Positive samples were further analyzed for H1N1, H3N2, and H1N2 antibodies with a hemagglutination inhibition test. Diagnostic samples for virus detection were subjected to influenza A M-gene-specific real-time RT-PCR and to pandemic influenza A H1N1-specific real-time RT-PCR. Positive samples were further analyzed with RT-PCRs designed for this purpose, and the PCR products were sequenced and sequences analyzed phylogenetically.

Results

In the blood samples from pigs in special health class farms producing replacement animals and in diagnostic blood samples, the first serologically positive samples originated from the period July–August 2008. In samples collected for disease monitoring, < 0.1%, 0% and 16% were positive for antibodies against influenza A H1N1 in the HI test in 2007, 2008, and 2009, respectively. Swine influenza A virus of avian-like H1N1 was first detected in diagnostic samples in February 2009. In 2009 and 2010, the avian-like H1N1 virus was detected on 12 and two farms, respectively. The pandemic H1N1 virus (A(H1N1)pdm09) was detected on one pig farm in 2009 and on two farms in 2010.

Conclusions

Based on our study, swine influenza of avian-like H1N1 virus was introduced into the Finnish pig population in 2008 and A(H1N1)pdm09 virus in 2009. The source of avian-like H1N1 infection could not be determined. Cases of pandemic H1N1 in pigs coincided with the period when the A(H1N1)pdm09 virus was spread in humans in Finland.  相似文献   

20.
A simple fluorometric method suitable for serial determinations is reported for simultaneous determination of vitamins A and E in serum. The solvents are pretreated chemically and by distillation. After shaking with ascorbic acid in ethanol, the vitamins are extracted from the serum samples with petroleum ether. The extract is used for the determination. The sensitivity of the method is 20 micrograms/100 ml for vitamin A and 0.04 micrograms/ml for vitamin E. Recovery rates are between 87 and 106%. The method was tested on a total of 160 serum samples of healthy animals belonging to 8 different species. The results showed good agreement with the so-called reference values published in the literature.  相似文献   

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