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1.
OBJECTIVE: To compare sensitivity and specificity of various polymerase chain reaction (PCR) assays for detection of Leptospira borgpetersenii serovar hardjo in bovine urine and to compare results of the optimal PCR assay with results of immunofluorescence, nucleic acid hybridization, and bacteriologic culture. ANIMALS: 6 heifers. PROCEDURE: Heifers were exposed to serovar hardjo type hardjo-bovis by conjunctival instillation of 10(6) leptospires on 3 successive days. Urine samples were collected before and after infection. Sensitivity and specificity of 5 PCR assays were compared, to determine the optimal assay for use with bovine urine samples. The optimal PCR assay was then compared with results of bacteriologic culture, nucleic acid hybridization, and immunofluorescence. RESULTS: A PCR assay with the best combination of specificity (100%) and sensitivity (91%) was selected for comparison with the other diagnostic tests. Sensitivity for nucleic acid hybridization was 55%, whereas sensitivity for bacteriologic culture and immunofluorescence was 89 to 93%. CONCLUSIONS AND CLINICAL RELEVANCE: Bacteriologic culture, PCR, and immunofluorescence were sensitive for detection of L borgpetersenii serovar hardjo type hardjo-bovis in urine specimens of cattle, but a single technique was not the most sensitive for each animal tested. Therefore, the use of 2 techniques in combination is warranted for maximal sensitivity for diagnosis. 相似文献
2.
Harkin KR Roshto YM Sullivan JT Purvis TJ Chengappa MM 《Journal of the American Veterinary Medical Association》2003,222(9):1230-1233
OBJECTIVE: To compare results of polymerase chain reaction (PCR) testing of urine samples, serologic testing, and bacteriologic culture of urine to determine prevalence of urinary shedding of leptospires in dogs. DESIGN: Serial case study. ANIMALS: 500 dogs evaluated serially without regard to health status. PROCEDURE: Urine samples were examined via PCR assay and bacteriologic culture for leptospires. Blood samples were analyzed for antibodies against serovars canicola, bratislava, pomona, icterohemorrhagiae, grippotyphosa, and hardjo. RESULTS: Titers > or = 1:100 against at least 1 serovar were detected in 104 (20.8%) dogs, and titers > or = 1:400 were detected in 41 (8.2%) dogs. High titers were detected most commonly to serovar grippotyphosa, followed by icterohemorrhagiae, canicola, pomona, bratislava, and hardjo. High titers to > 1 serovar were detected in 14 dogs. A positive PCR assay result was obtained in 41 (8.2%) dogs, only 9 of which had a titer > or = 1:100. Leptospires were not cultured from the urine of any dog. Only 4 dogs had clinical leptospirosis. Overall disease prevalence was 0.8% for the 6-month evaluation period. Compared with PCR assay, serologic testing for predicting shedding had a sensitivity of 22%, specificity of 79%, positive predictive value of 9%, and negative predictive value of 92%. CONCLUSIONS AND CLINICAL RELEVANCE: Irrespective of health status, 8.2% of dogs were shedding pathogenic leptospires. Serologic testing was a poor predictor of urinary shedding. Clinically normal dogs that shed leptospires may pose a zoonotic risk to their owners. 相似文献
3.
Experimental infection of calves with Leptospira interrogans serovar hardjo: conjunctival versus intravenous route of exposure 总被引:1,自引:0,他引:1
Eight-month-old calves, housed under maximum isolation, were exposed to pathogenic Leptospira interrogans serovar hardjo by the conjunctival route or IV. One calf served as an unexposed control. Infection was monitored serologically (microscopic agglutination test and enzyme-linked immunosorbent assay; ELISA) and by leptospiral culture isolation from periodic urine samples and from the kidneys, epididymides, and aqueous humor collected at slaughter. Microscopic agglutination test titers of greater than or equal to 1:40 were detected among all IV exposed calves at postinoculation day (PID) 7 and among conjunctival exposed calves at PID 14. By ELISA, all IV exposed calves were positive by PID 3, whereas conjunctival exposed calves were positive at PID 14. The ELISA was more sensitive for the detection of antibodies against leptospires in cattle. Leptospires were isolated from the urine of 4 calves and from the kidney of 3 calves exposed by the conjunctival route, but not from IV exposed calves. The results indicated that the conjunctival route of exposure was a more natural and successful route for experimental infection of cattle with serovar hardjo. 相似文献
4.
We evaluated the use of low-stringency single specific primer PCR (LSSP-PCR) for genetically typing Leptospira directly from urine samples of cattle with clinical suspicion of leptospirosis. Urine samples obtained from 40 cattle with clinical suspicion of leptospirosis were amplified by specific PCR using the following primers: Internal 1/Internal 2 and G1/G2. The internal primers were designed from the gene sequence of the outer membrane lipoprotein Lip32 from Leptospira kirschneri, strain RM52. The PCR products were amplified with these two pairs of primers, which had approximately 497 and 285bp, respectively, and were subsequently used as a template for LSSP-PCR analysis. The genetic signatures from the leptospires which were present in the urine samples allowed us to make a preliminary identification of the leptospires by comparing the LSSP-PCR profiles obtained directly from urine samples with those from reference leptospires. The LSSP-PCR profiles obtained with the Internal 1 primer or with the G1 primer allowed the grouping of the leptospires into serogroups. LSSP-PCR was found to be a useful and sensitive approach capable of identifying leptospires directly from biological samples without the need for prior bacterial isolation. In conclusion, the LSSP-PCR technique may still be helpful in discriminating serogroups of Leptospira from different animal reservoirs, since the early identification of carrier animals and information on the shedding state are crucial to prevent the spread of leptospiral infection to other animals and humans. 相似文献
5.
Bomfim MR Barbosa-Stancioli EF Koury MC 《Veterinary journal (London, England : 1997)》2008,178(2):251-256
A nested polymerase chain reaction (PCR) using primers from the LipL32 sequence of Leptospira spp. was used to detect shedding of pathogenic leptospires in urine from naturally infected cattle. Amplicons (497bp) were obtained from 21 pathogenic reference serovars belonging to four species (L. interrogans, L. borgpetersenii, L. santarosai, L. kirschneri). DNA was amplified from 26/30 urine samples taken from cattle with suspected leptospirosis and from leptospires cultivated from 10 of these samples. The limit of detection of DNA in the clinical samples was 200pg and the nested PCR detected all pathogenic reference serovars of Leptospira spp. tested. No PCR products were amplified using DNA from other common bacterial species from the bovine urogenital tract or urine, or from the non-pathogenic L. biflexa Andamana serovar. The nested PCR exhibited high specificity and sensitivity for detection of pathogenic serovars in urine from cattle. 相似文献
6.
Shedding and Seroprevalence of Pathogenic Leptospira spp. in Sheep and Cattle at a New Zealand Abattoir
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F. Fang J. M. Collins‐Emerson A. Cullum C. Heuer P. R. Wilson J. Benschop 《Zoonoses and public health》2015,62(4):258-268
A cross‐sectional study was carried out on sheep and cattle slaughtered at a New Zealand abattoir from September to November 2010 to investigate the supplier‐specific shedding rate, renal carriage rate and seroprevalence of leptospires. In the 2008/2009 season, this abattoir experienced three human leptospirosis cases from 20 staff, of which two were hospitalized. Urine, kidney and blood samples were collected from carcasses of 399 sheep (six suppliers, 17 slaughter lines) and 146 cattle (three suppliers, 22 slaughter lines). The urine and kidney samples were tested by quantitative real‐time PCR (qPCR), while serum samples (from coagulated blood samples) were tested by microscopic agglutination test (MAT). In total, 27% (73/274; 95% CI: 18–37) of urine samples tested positive by qPCR. Species‐specific shedding rates (prevalence of positive urine qPCR) were 31% (95% CI: 17–48) for sheep and 21% (95% CI: 14–30) for cattle. For 545 kidney samples tested, 145 were qPCR positive (27%; 95% CI: 17–39). The average prevalence of kidney qPCR positivity was 29% (95% CI: 17–45) for sheep and 21% (95% CI: 15–28) for cattle. Three hundred and thirty of 542 sampled sheep and cattle had antibodies against Leptospira borgpetersenii serovar Hardjobovis (Hardjobovis) and/or Leptospira interrogans serovar Pomona (Pomona), based on reciprocal MAT titre ≥1 : 48 (overall seroprevalence of 61%; 95% CI: 48–73). Seroprevalence was 57% (95% CI: 40–72) for sheep and 73% (95% CI: 59–83) for cattle. Among the seropositive animals, 41% (70/170; 95% CI: 30–54) were shedding (tested positive by urine qPCR) and 42% (137/330; 95% CI: 30–54) had renal carriage (tested positive by kidney qPCR). Some risk management options for abattoirs or farms to prevent human leptospirosis infections include vaccination of maintenance hosts, the use of personal protective equipment, and the application of urine qPCR to detect shedding status of stock as surveillance and as an alert. 相似文献
7.
Dunn JR Kaneene JB Grooms DL Bolin SR Bolin CA Bruning-Fann CS 《Journal of the American Veterinary Medical Association》2005,226(3):429-435
OBJECTIVE: To determine whether cattle testing positive for Mycobacterium avium subsp paratuberculosis as determined by microbial culture of feces or antibody ELISA were more likely to have false-positive responses on the caudal fold tuberculin (CFT) test or interferon-gamma (IFN-gamma) assay for Mycobacterium bovis than cattle testing negative for M paratuberculosis. ANIMALS: 1043 cattle from 10 herds in Michigan. PROCEDURE: Feces and blood samples for plasma were collected from cattle > or =24 months old on the day the CFT test was read. Fecal samples were submitted for microbial culture for M paratuberculosis. Plasma samples were tested for antibody against M paratuberculosis, and IFN-gamma after stimulation with purified protein derivative tuberculin from M bovis or M avium. RESULTS: Of 1043 cattle, 180 (17.3%) had positive CFT test results (suspects) and 8 (0.8%) had positive IFN-gamma assay results after stimulation with purified protein derivative tuberculin from M bovis. Forty-five (4.3%) and 115 (11.0%) cattle tested positive for M paratuberculosis as determined by microbial culture of feces and antibody ELISA, respectively. Cattle with positive responses for M paratuberculosis appeared to have an increased likelihood of false-positive results on the CFT test, although this association was not significant. CONCLUSIONS AND CLINICAL RELEVANCE: No significant association was detected among cattle testing positive for M paratuberculosis as determined by microbial culture of feces and antibody ELISA and positive CFT test and IFN-gamma assay results for M bovis. 相似文献
8.
Fearnley C Wakeley PR Gallego-Beltran J Dalley C Williamson S Gaudie C Woodward MJ 《Research in veterinary science》2008,85(1):8-16
A LightCycler real-time PCR hybridization probe-based assay that detects a conserved region of the16S rRNA gene of pathogenic but not saprophytic Leptospira species was developed for the rapid detection of pathogenic leptospires directly from processed tissue samples. In addition, a differential PCR specific for saprophytic leptospires and a control PCR targeting the porcine beta-actin gene were developed. To assess the suitability of these PCR methods for diagnosis, a trial was performed on kidneys taken from adult pigs with evidence of leptospiral infection, primarily a history of reproductive disease and serological evidence of exposure to pathogenic leptospires (n=180) and aborted pig foetuses (n=24). Leptospire DNA was detected by the 'pathogenic' specific PCR in 25 tissues (14%) and the control beta-actin PCR was positive in all 204 samples confirming DNA was extracted from all samples. No leptospires were isolated from these samples by culture and no positives were detected with the 'saprophytic' PCR. In a subsidiary experiment, the 'pathogenic' PCR was used to analyse kidney samples from rodents (n=7) collected as part of vermin control in a zoo, with show animals with high microagglutination titres to Leptospira species, and five were positive. Fifteen PCR amplicons from 1 mouse, 2 rat and 14 pig kidney samples, were selected at random from positive PCRs (n=30) and sequenced. Sequence data indicated L. interrogans DNA in the pig and rat samples and L. inadai DNA, which is considered of intermediate pathogenicity, in the mouse sample. The only successful culture was from this mouse kidney and the isolate was confirmed to be L. inadai by classical serology. These data suggest this suite of PCRs is suitable for testing for the presence of pathogenic leptospires in pig herds where abortions and infertility occur and potentially in other animals such as rodents. 相似文献
9.
10.
A study was conducted in 2008 to determine the prevalence of Anaplasma and Babesia infections in cattle in the Puntarenas Province of Costa Rica. Blood samples were taken from a total of 449 cattle during the month of March at 30 farms in the region of Espiritu Santu, Costa Rica. Commercially available enzyme-linked immunosorbent assays (ELISA) were used to determine presence of antibodies to Babesia bigemina and Anaplasma marginale, and real-time PCR was used to determine the presence of DNA from the disease-causing organisms. The ELISA results indicated that 87.5% of the cattle sampled were positive for antibodies to A. marginale, while 59.1% were positive for antibodies to B. bigemina. The real-time PCR results showed that 235 cattle were carrying A. marginale DNA (56.9%), 6 with B. bigemina DNA (1.34%), and 2 with B. bovis DNA (0.45%). 相似文献
11.
Hugh Y Cai Geoff Hornby Douglas W Key Maria R Osuch M Grant Maxie 《Journal of veterinary diagnostic investigation》2002,14(2):164-168
A multiplex polymerase chain reaction (PCR) method using primer sets of G1/G2 and B64-I/B64-II was validated to detect pathogenic leptospira serovars from canine urine samples. The PCR method was found to be specific and sensitive with a detection limit of 100 cells of Leptospira icterohaemorrhagiae per milliliter of urine. The primer set previously designated and erroneously transcribed B64-I/B64-II amplified a DNA fragment of 352 base pairs from Leptospira grippotyphosa and Leptospira sejroe but not from Leptospira autumnalis, Leptospira bratislava, Leptospira canicola, Leptospira hardjo, Leptospira icterohaemorrhagiae, and Leptospira pomona. From 100 diagnostic canine urine samples, 5 were found positive for Leptospira grippotyphosalsejroe with a PCR product of 352 base pairs and 6 were positive for other pathogenic leptospira serovars with a PCR product of 285 base pairs. One 285-base pair product was sequenced and found to be 99.3% homologous to the G1/G2 PCR fragment sequence reported previously. All 352-base pair PCR products of clinical samples and pure cultures of L. grippotyphosa and L. sejroe were sequenced. The 352-base pair fragment sequences of L. grippotyphosa and L sejroe were identical. Only 2 base pairs were found different between the sequences from pure cultures and those from clinical samples. Serum samples from 3 positive cases that generated a PCR product of 352 base pairs were tested by the microscopic agglutination test, and 2 were found to be positive for L. grippotyphosa (1:10,240 and 1:5,120), 1 was positive for L. grippotyphosa (1:320) or L. icterohaemorrhagiae (1:320). The results of this study suggest that the multiplex PCR with the primer set G1/G2 and the erroneously transcribed B64-I/B64-II may be able to differentiate L. grippotyphosa or L. sejroe from other pathogenic leptospira serovars commonly tested for in Canadian diagnostic laboratories. 相似文献
12.
Isolation of Leptospira hardjo from kidneys of Ontario cattle at slaughter. 总被引:3,自引:3,他引:0
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Kidneys from 117 cattle from 110 Ontario farms were examined at slaughter for leptospires. Leptospira hardjo (hardjo-bovis A) was isolated from 11 kidneys and L. kennewicki from one. The isolations were all made (12/89, 13.5%) from beef cattle from feedlots, no isolates being obtained from dairy or beef cattle from extensive farms (0/28). Isolations were only made from cattle with antibody titers (greater than or equal to 20) against the serovars recovered. Isolation was more sensitive than immunofluorescence in identifying leptospira, particularly in animals with low antibody titers against L. hardjo. Leptospira were isolated from two kidneys with multiple gross lesions of focal nephritis, but there was no correlation between the presence of scanty kidney lesions and isolations of leptospira. Leptospira hardjo infection appears to be common in Ontario feedlot cattle. 相似文献
13.
Surya Sankar Hiron M. Harshan S.R. Somarajan S.K. Srivastava 《Research in veterinary science》2010,88(3):375-378
A recombinant leptospiral lipoprotein, LigB, was evaluated for use in the diagnosis of bovine leptospirosis by enzyme-linked immunosorbent assay (rLigB IgG ELISA). The standard reference test (Microscopic agglutination test, MAT) of 200 serum samples from cattle suspected of leptospirosis showed that 95 (47.5%) samples had positive agglutination titres, which ranged from 100 to 1600. In rLigB IgG ELISA, 49% of the samples were positive. Sensitivity of IgG ELISA for 95 bovine sera, which had MAT titres of greater than or equal to 100, were 100%. ELISA showed a specificity of 97.1% with 105 bovine sera, which were negative at a 1:50 dilution in MAT for Leptospira interrogans serovars. The results of ELISA and MAT correspond very good. When analytical specificity of IgG ELISA was evaluated using bovine serum samples from animals showing the serum antibodies to other pathogens, no cross-reaction was observed. Thus the recombinant LigB IgG ELISA can be used instead of the MAT as an aid to the diagnosis of bovine leptospirosis. 相似文献
14.
Alt DP Zuerner RL Bolin CA 《Journal of the American Veterinary Medical Association》2001,219(5):636-639
OBJECTIVE: To evaluate antibiotics for treatment of cattle with leptospirosis caused by Leptospira borgpetersenii serovar hardjo. DESIGN: Randomized controlled trial. ANIMALS: 42 healthy mixed-breed cattle. PROCEDURE: Cattle were inoculated via conjunctival instillation with L. borgpetersenii serovar hardjo. After infection and urinary shedding of L. borgpetersenii were confirmed, cattle were treated with various antibiotics. To determine effectiveness of antibiotic treatment, urinary shedding of L. borgpetersenii was monitored for 4 to 6 weeks after administration of antibiotics, using darkfield microscopic examination, microbial culture, immunofluorescence testing, and a polymerase chain reaction assay. RESULTS: All inoculated cattle developed leptospirosis and shed leptospires in their urine. The following antibiotic treatments resulted in elimination of urinary shedding of leptospires: a single injection of oxytetracycline (20 mg/kg 19 mg/lb] of body weight, IM), tilmicosin (10 mg/kg [4.5 mg/lb], SC), or a combination product that contained dihydrostreptomycin-penicillin G (25 mg/kg [11.4 mg/lb], IM) or multiple injections of ceftiofur sodium (2.2 or 5 mg/kg [1 or 2.3 mg/lb], IM, once daily for 5 days, or 20 mg/kg, IM, once daily for 3 days). CONCLUSIONS AND CLINICAL RELEVANCE: Successful resolution of leptospirosis in cattle by administration of dihydrostreptomycin-penicillin G confirms results obtained by other investigators. Three other antibiotics (oxytetracycline, tilmicosin, and ceftiofur) also were effective for resolving leptospirosis and may be useful substitutes for dihydrostreptomycin, an antibiotic that is no longer available for use in food-producing animals in the United States. Cost, safety, and withdrawal times of these various treatment options need to be considered. 相似文献
15.
AD Mackay 《New Zealand veterinary journal》2013,61(6):281-288
Abstract AIM: To find evidence for localisation in the uterus, and fetal infection, of Leptospira spp. in farmed deer in the lower North Island of New Zealand during and shortly after the breeding season. METHODS: Between February and July 2008, 116 blood samples, 120 kidneys, 120 uteri and 27 fetuses were collected from 120 mixed-age hinds from lines from nine farms, at a deer slaughter premises. Serum samples were tested for antibodies against Leptospira borgpetersenii serovar Hardjo-bovis and Leptospira interrogans serovar Pomona, using the microscopic agglutination test (MAT). For both serovars, a titre of >1:48 was considered positive. Samples from kidneys, uteri and fetal tissue were subjected to bacterial culture, using Ellinghausen-McCullough-Johnson-Harris (EMJH) medium, and real-time PCR, using DNA gyrase subunit B gene primers. RESULTS: Thirty-four of 116 (29.3%) serum samples were positive for serovar Hardjo-bovis, and 13 (11.2%) for serovar Pomona. Seven of 120 kidneys were positive for serovar Hardjo-bovis by culture, and five of these, but no others, were positive by real-time PCR. Of 120 uteri, none was culture- or PCR-positive. None of 27 fetal samples was culture-positive but one was positive by real-time PCR. The dam of the PCR-positive fetus was culture-negative from the kidney, but had an MAT titre of 1:192 for Hardjo-bovis. CONCLUSIONS: Attempts to isolate Leptospira spp. from the genital tracts and early fetuses of farmed deer were unsuccessful. However, molecular evidence suggested fetal infection in one case. This finding justifies further study of the role of leptospires in the genital tract and fetus and its association with reproductive loss in farmed deer. 相似文献
16.
R J Chappel B Adler S A Ballard S Faine R T Jones B D Millar J A Swainger 《Veterinary microbiology》1985,10(3):279-286
An enzymatic radioimmunoassay (ERIA) has been developed for detecting Leptospira interrogans serovar pomona in porcine urine. Four grower pigs were experimentally infected with serovar pomona. A total of 39 urine samples was collected, and ERIA was compared with dark ground microscopy (DGM) and culture for demonstrating leptospiruria. Of 20 samples positive by at least one technique, leptospires were detected by ERIA in 14, by culture in 16 and by DGM in 13. ERIA, unlike the other 2 methods, was suitable for use with urine which had been stored frozen for several months. 相似文献
17.
Lordrick Alinaitwe Clovice Kankya Kathryn J. Allan Sabrina Rodriguez‐Campos Paul Torgerson Anou Dreyfus 《Zoonoses and public health》2019,66(6):636-646
Leptospirosis is a zoonotic bacterial disease reported worldwide. In Uganda, seropositivity has been reported in both humans and domesticated animals, including cattle. However, it remains unknown whether cattle are shedding leptospires and thus acting as potential source for human leptospirosis. We conducted this cross‐sectional study in two cattle abattoirs in Kampala, Uganda between June and July 2017. Kidney and urine samples from 500 cattle sourced from across the country were analysed by real‐time PCR to establish the prevalence of Leptospira‐positive cattle and risk of exposure to abattoir workers. The species of infecting Leptospira was determined by amplification of secY gene and compared to reference sequences published in GenBank. Of 500 cattle tested, 36 (7.2%) had Leptospira DNA in their kidneys (carriers), 29 (5.8%) in their urine (shedders); with an overall prevalence (kidney and/or urine) of 8.8%. Leptospira borgpetersenii was confirmed as the infecting species in three cattle and Leptospira kirschneri in one animal. Male versus female cattle (OR = 3, p‐value 0.003), exotic versus local breeds (OR = 21.3, p‐value 0.002) or cattle from Western Uganda (OR = 4.4, p‐value 0.001) and from regions across the border (OR = 3.3, p‐value 0.032) versus from the central region were more likely to be Leptospira‐positive. The daily risk of exposure of abattoir workers to ≥1 (kidney and/or urine) positive carcass ranged from 27% (95% credibility interval 18.6–52.3) to 100% (95% CI 91.0–100.0), with halal butchers and pluck inspectors being at highest risk. In conclusion, cattle slaughtered at abattoirs in Uganda carry and shed pathogenic Leptospira species; and this may pose occupation‐related risk of exposure among workers in these abattoirs, with workers who handle larger numbers of animals being at higher risk. 相似文献
18.
Wilson PR McGhie J Marshall RB Audigé LJ Collins-Emerson J Quankai W Alley MR 《New Zealand veterinary journal》1998,46(4):131-139
AIMS: Slaughterhouse and on-farm surveys were undertaken to investigate some aspects of leptospirosis (Leptospira interrogans) in farmed deer in the lower North Island of New Zealand. METHODS: Blood samples and kidneys were collected at slaughter from 601 l-year and older red and red X Wapiti stags and 21 adult hinds from 53 farms (10 or 12 deer per farm). Serum samples were analysed for up to seven Leptospiral serovars. Gross and histological examinations of kidneys were undertaken. Kidneys from 202 deer were cultured for leptospires. A follow-up postal questionnaire (68% response) indicated one herd had been vaccinated prior to the survey. Serological analyses were also carried out on serum bank samples from a previous on-farm survey involving male and female weaner, yearling and adult red deer from 16 commercial deer farms in March and November. RESULTS: Serological reactions at titres > or = 96 to serovar hardjo were present in 73.6%, pomona in 41.5%, copenhageni in 11.3% and tarassovi in 15.1% of farms from the slaughterhouse survey. Antibodies to serovars australis, ballam and balanica were present in three, one and four of six herds studied, respectively. Titre prevalence to hardjo was higher than that of pomona and other serovars within farms. Cultures for Leptospira were positive in 10 stags from six lines with similar prevalence across age groups. Histological examination showed many gross lesions were associated with mild interstitial cellular infiltration characteristic of subclinical Leptospiral infections. Some sections from culture-positive kidneys contained spirochetes in renal tubules. The on-farm survey showed a 10-30% within-herd prevalence of pomona and hardjo titres in 56% of 3-month-old deer herds, but by 11 months of age, 100% of herds were titre-positive with high prevalences to one or both serovars. Concurrently, herds of 1-year-old and adult deer on the same farms were all seropositive. CONCLUSION: This study has shown that Leptospiral infections are common in farmed deer in the survey area. 相似文献
19.
Gutiérrez SE Dolcini GL Arroyo GH Rodriguez Dubra C Ferrer JF Esteban EN 《American journal of veterinary research》2001,62(10):1571-1577
OBJECTIVE: To develop a blocking ELISA for detection of bovine leukemia virus (BLV) antibodies that is comparable to a radioimmunoprecipitation (RIP) assay, to evaluate use of this ELISA for identification of BLV-infected herds, and to develop a polymerase chain reaction (PCR) assay for direct diagnosis of infection with BLV. SAMPLE POPULATION: Serum samples and pooled bulk-tank milk samples from cattle. PROCEDURE: The blocking ELISA was developed, using BLV gp51 as antigen, captured by a selected bovine polyclonal serum. A nested PCR was conducted with primers specific for a segment of the pol region of the BLV genome. RESULTS: Sensitivity and specificity of the ELISA were comparable to those of the RIP assay. Use of the ELISA on pooled milk samples allowed identification of herds in which prevalence of BLV infection among lactating cows was as low as 2.5%. Pooled milk samples from BLV-free herds did not react in the ELISA. All cattle that had positive results for the nested PCR had BLV antibodies, but cattle with consistantly low antibody titers required examination of sequential DNA samples to detect viral sequences. None of the 63 antibody-negative cattle had positive results for the PCR. CONCLUSIONS AND CLINICAL RELEVANCE: This ELISA is a highly specific and sensitive assay for the detection of BLV antibodies in serum and milk samples of cattle. Examination of pooled milk samples with the ELISA provides a reliable, practical, and economic procedure for identification of BLV-infected herds. The nested PCR also constitutes a specific procedure for direct diagnosis of infection with BLV. 相似文献
20.
Sakoda Y Naito M Ito M Ito Y Isoda N Tanaka T Umemura T Kida H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(7):955-958
Leptospira interrogans serovar Manilae strain UP-MMC was inoculated into miniature pigs to assess its pathogenicity. Leptospires were recovered from the whole blood, kidneys, and livers in the acute phase without showing any clinical signs. Under immunosuppressive conditions by dexamethasone, leptospires were recovered from the kidneys and their genes were detected from the urine in the chronic phase. These results indicate that leptospires persisted in the kidneys until the chronic phase, and excretion of leptospires in the urine was enhanced under immunosuppressive conditions, resulting in horizontal transmission among pigs on farms. 相似文献