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1.
Hamster inoculation, medium inoculation and microscopical methods were used for the detection of Leptospira interrogans serovar pomona organisms in bovine urine. Urine samples were collected from both naturally and experimentally infected animals during the period when leptospiruria could be expected.

Inoculation into hamsters of 0.5 ml of urine from each of 14 samples resulted in 5 positives. The inoculation of 0.025 ml of the same sample into semi-solid EMJH medium gave 10 positives. From a cumulative total of 46 urine samples, 28 were positive using dark-ground microscopy of centrifuged urine deposits and 35 were positive using media inoculation. A cumulative total of 126 urine samples were examined, after centrifugation, under dark-ground microscopy and leptospirae were detected in 60.

Fluorouracil (100 mg/ml) proved to be beneficial for successful isolations in media, whereas neomycin (5 mg/ml) did not.  相似文献   

2.
Investigations were carried out in 1975, 1976 and 1977 in 16 dairy herds where leptospiral abortions were suspected and in five other herds where clinical disease was not present. Both Leptospira interrogans serovars pomona and hardjo were isolated from cattle in herds with leptospirosis, but only pomona was recovered from those that had aborted. There was no evidence that hardjo caused clinical disease in dairy cattle in the Waikato district.

It was found that 73% of the cows that aborted and 19% of other animals in the same herds had microscopic agglutination test titres to pomona of 1:2 000 or greater. By contrast, only 2% of cattle in herds without clinical evidence of leptospirosis had such titres. One cow retained a titre of 1:2 000 or greater to pomona for 7 months; titres of this order had a shorter duration in other cows. Leptospiruria occurred in 50% of cows that had aborted and in 9% of in-contact cows in the same herds. Only 0.7% of cows had leptospiruria in the herds with no clinical disease. Ten of 35 cows shedding pomona still had leptospiruria one month later.

It was concluded that clinical leptospirosis should be diagnosed by testing a sample of the herd, rather than just individual cows, because of the variability and persistence of leptospiruria and serological titres in cows with and without clinical signs. Although hardjo is common in cattle in the Waikato district, it was not found to cause abortion in cattle.  相似文献   

3.
The results of combined epidemiological, clinical, serological, bacteriological and histopathological studies following an outbreak of disease caused by L. pomona on a farm stocked with cattle, sheep, pigs, goats and horses maintained for experimental purposes, are reported.

The incidence of infection was high in horses, cattle and pigs. A few low titres were seen in sheep. The goats were not infected. Apart from a single bovine abortion all the clinical symptoms observed occurred in pregnant sows. Seven of these aborted or gave birth to stillborn pigs within a six week period.

Fifteen species of wildlife were trapped or shot on the farm during the year following the outbreak. L. pomona was isolated from four skunks and a porcupine. Epidemiological studies indicated that wildlife reservoir hosts were the primary source of infection for the domestic livestock.

Leptospiruria and the serological response were studied in a group of eight infected sows. Microscopic agglutination titres of 102 or less could not be associated with leptospiruria and the duration of leptospiruria was found to range from a few weeks to over two years in individual sows. Direct dark-field examination of urine proved superior to guinea-pig inoculation as a method of detecting leptospiruria and it is suggested that the former technique could be adopted with advantage as a routine aid to diagnosis.

  相似文献   

4.
A trial was conducted to assess the efficacy of a single intra-muscular injection of 25mg per kg of streptomycin in stopping leptospiruria in growing pigs exposed to a natural infection.

The investigation involved 54 pigs, 10–12 weeks old, that were free of serological evidence of infection with serotypes pomona or tarassovi at the start of the trial. The animals were kept in pens in a finishing house through which flowed effluent from other pens containing infected pigs. Urine samples were collected from each pig three times weekly until it was slaughtered for bacon at 20–24 weeks old.

Leptospiruria was first detected between 16 and 74 days after exposure. Twenty-one of 37 pigs which showed leptospiruria were injected with streptomycin. Leptospirae were detected in 11 of 185 (5.9%) subsequent urine samples from these 21 treated animals and serotype pomona was cultured from the kidney of 1 of the 10 animals that were examined from this group at slaughter.

Following the initial detection of leptospiruria in the 16 un-treated pigs, 179 of 211 (84.8%) subsequent urine samples con-tained leptospirae, which were also cultured from the kidneys of 4 out of 11 of these animals.  相似文献   

5.
Fourteen of 24 pigs were immunised with repeated injections of killed cultures of Leptospira serotype pomona administered over a three-week period. The remaining 10 pigs served as controls.

Five days after immunisation all pigs were exposed to the same natural infection by being housed together for 12 weeks in a pen which received effluent from other pens containing infected pigs.

Neither leptospiruria nor kidney lesions were detected in any of the immunised pigs. In contrast, leptospiruria and kidney lesions occurred in all the controls.

The development of serum agglutinins after immunisation and before exposure suggested that this could be used as an index of adequate antigenic stimulation.  相似文献   

6.
The efficacy of the hardjo component of a hardjo-pomona vaccine was evaluated in yearling heifers under conditions of natural challenge in a commercial dairy herd when endemic hardjo infection was present. Eight heifers received 2 doses of vaccine 4 weeks apart and were run with 10 unvaccinated heifers for a period of 56 weeks. Results from the culture of urinesamples showed that the vaccine either prevented leptospiruria to a significant degree (P<0.05) or, if it developed, greatly shortened its duration (P<0.01). Leptospires were cultured on an average of 5 occasions (range 3 to 8) from each of the infected controls and on only one occasion each from 2 of the vaccinates. Fifty one to 56 weeks after commencement of the trial, 9 of the unvaccinated animals were excreting leptospires while none of the vaccinates were leptospiruric at that time.

It is concluded that an appropriate vaccination programme could prevent the maintenance of hardjo infection in the herd.  相似文献   

7.
1. Successful invasion by nematode parasites is associated with several factors including egg hatching at the right time in their hosts. To determine a simple and appropriate medium for culture and egg hatching of the highly pathogenic species of the Acuariidae family, Cheilospirura hamulosa were cultured in three different media. In addition the viability of C. hamulosa eggs was determined after storage in frozen infected gizzards.

2. Eggs removed from the uteri of the female worms in infected gizzards were pooled and washed in distilled water and screened under a stereo dissecting microscope. Eggs were counted and cultured in three different media, nutrient agar, normal saline 0.9% and Bearman, at room temperature. Additionally, 10 infected gizzards were kept at ?20°C for 2 and 8 months.

3. After 4–5 d there had been no growth in the nutrient agar medium, whereas 11% of the cultured eggs in the Bearman medium contained larvae 2–3 d after culturing. In 0.9% normal saline medium the two polar knobs appeared on the two poles of the eggs at 2 d post cultivation, and 74% of the eggs contained a larva on the third day. Mature larvae gradually exited from the eggs.

4. Eggs collected from female worms in gizzards frozen at ?20°C were cultured in the same three culture media at room temperature. Larvae were visible in the eggs after 2–3 d in the Bearman and 0.9% normal saline media and hatched thereafter.

5. The 0.9% normal saline medium is recommended for egg hatching and cultivation of C. hamulosa due for simplicity, efficacy and cost effectiveness. Moreover, freezing of the infected gizzards at ?20°C is proposed for long-term storage of the eggs.  相似文献   

8.
Leptospira interrogans serovars pomona, hardjo and tarassovi were each used to inoculate 6 cattle. Three-hundred and ninety-nine sera collected from the inoculated animals and from a control group over a 3-month period were tested using the microscopic agglutination test (MAT) and the enzyme-linked immunosorbent assay (ELISA). Leptospiruria was monitored by microscopic examination and culture. The ELISA detected specific IgM antibody against the serovars in all infected cattle 1 week after inoculation. This IgM antibody persisted in most of the animals for 3-5 weeks. Specific IgG antibody appeared at the same time or just after IgM, but persisted for much longer. Levels of antibody detected by the ELISA and the MAT did not correlate with each other, nor with the periods of leptospiruria found in the infected cattle.  相似文献   

9.
SUMMARY The efficacy of a vaccine against Leptospira interrogans serovar pomona and Leptospira interrogans serovar hardjo was evaluated in a group of dairy heifers that were serologically negative at the time of vaccination and later subjected to natural challenge with L. interogans serovar hardjo. Thirty-nine heifers were vaccinated twice, at a one-month interval, with a commercially prepared bivalent vaccine, while 43 unvaccinated heifers were used as controls. After vaccination, microscopic agglutination (MA) titres of serums to L. interrogans serovar hardjo ranged from 32 to 512, and those to L. interrogans serovar pomona ranged from 32 to 2048. Titres resulting from vaccination were short-lived and after the first vaccination the serums of 95% of vaccinated heifers did not react in the MA test by 24 weeks. The first indication of infection in the heifers was noted at week 6, and by week 16, elevated MA titres (≥128) to L. interrogans serovar hardjo had occurred in 62% of unvaccinated heifers and had increased to 85% by week 24. At week 18, 18% of the vaccinated heifers and 56% of the unvaccinated heifers had leptospiruria (p<0.01); after 22 weeks, 13% of the vaccinated heifers and 58% of the unvaccinated heifers showed evidence of leptospiruria (p<0.01).  相似文献   

10.
Anaplasma marginale was propagated in a tick cell line derived from Dermacentor variabilis embryos. The rickettsial organism was identified and monitored in culture by transmission electron microscopy and the indirect immunofluorescence technique, using specific monoclonal antibodies. Inoculation of the embryonic tick cell line with midguts of infected adult ticks (culture 1), nymphal ticks (culture 2) and adult ticks that were infected as nymphs and dissected as adults (culture 3) resulted in 3 continuous cultures of A marginale. Culture 1 had been maintained through 22 passages over a 11-month period; cultures 2 and 3 had been maintained for 18 passages over a 9-month period. Growth of A marginale in the cell line began in the area of the nuclear membrane at approximately 4 days after inoculation or transfer. Thereafter, the organisms were observed in inclusions scattered throughout the cytoplasm of the host cells. Maximal growth of the organism occurred at 7 to 14 days, after which numbers of inclusions rapidly decreased to minimal or undetectable levels. The organism began new cycles of growth with each 1:5 to 1:10 split and transfer of the host cells. Electron microscopy of recently infected cells revealed a morphology of the organism that closely resembled that observed in marginal bodies of infected erythrocytes. After several passages, A marginale organisms had a varied morphology and resembled the organism described in midgut cells of naturally infected ticks.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
The serological, bacteriological and histopathological characteristics of experimental infection with serovar balcanica in possums (Trichosurus vulpecula) are described, and the possum is shown to be a potential maintenance host for this organism. Serum agglutination titres were maintained at almost constant levels for longer than a year, and leptospiruria was present in 50% of animals for a similar period. A paradoxical reaction to hardjo antigen was found in sera from all possums infected with balcanica.

Comparative studies were conducted using recently isolated field strains of serovars hardjo and ballum. Young possums seronegative to Hebdomadis group titres were insusceptible to challenge with hardjo, and the pathogenesis of ballum infection was characteristic of leptospiral infection in an accidental host.

The occurrence of antibodies in the urine of possums infected with balcanica is also described.  相似文献   

12.
SUMMARY Leptospirosis associated with probable L. hardjo infection was investigated in a dairy herd in a coastal district of Western Victoria. Thirty-six of 110 cows suffered leptospiruria and mastitis characterised by flaccid udders and abnormal milk. One of two media used enabled the isolation of the organism from infected guinea pigs inoculated with fresh urine. Microscopic agglutination titres to L. hardjo were elevated during the outbreak. There was an associated human infection.  相似文献   

13.
Abstract

Largemouth bass virus (LMBV), a recently discovered iridovirus found in the eastern United States, is usually detected by isolation in cell culture. Although LMBV will replicate in several cell lines, optimal cell culture methods for the detection of this virus have not been determined. We tested inoculation method, adsorption time, incubation temperature, and various cell lines to determine the conditions that would provide the most sensitive cell culture assay for LMBV. The optimal inoculation procedure tested was to remove the culture medium from the culture well before the addition of the inoculum, and the optimal adsorption procedure tested was to allow the virus to adsorb for 40 min while the plates were on an orbital shaker. Following inoculation, incubation at 30°C resulted in a higher number of viral plaques than incubation at 25°C or 32°C. Four cell lines (bluegill fry (BF-2), fathead minnow (FHM), epithelioma papulosum cyprini, and channel catfish ovary cells) inoculated with LMBV had similar susceptibility to infection. Similar percentages of LMBV-positive samples were detected in BF-2 and FHM cell cultures inoculated with homogenized organ samples from largemouth bass Micropterus salmoides; however, the use of two cell lines increased the number of infected samples discovered. A blind passage also increased the number of positive samples detected in cell culture. Subcultivation to confirm virus-positive samples was useful for reducing false-positive results.  相似文献   

14.
Leptospira interrogans serovar pomona was found to survive for at least 42 days in a typical New Zealand soil under simulated winter field conditions. The soil was markedly acidic with a pH of 5.5 and survival times were not reduced even when its water content was only 23%. The values of both these parameters are considerably less than previously recorded for the survival of leptospires in soil. Two methods were used to recover leptospires from the soil microflora. One was the culture of a membrane filtrate in EMJH media with or without contaminant-suppressing additives and the other was the direct inoculation of soil-washings into hamsters. Both techniques proved to equally sensitive. It was estimated that following the addition of 5 X 10(8) leptospires to the soil samples less than 2 X 10(4) were present after six weeks.  相似文献   

15.
Abstract

Extract

Sir, — We were interested to see the results of the trial conducted by Flint and Liardet (2) Flint, S. H. and Liardet, D. M. 1981. Leptospira interrogans serovar hardjo vaccination against Leptospira interogans serovar balcanica infection. N.Z. vet. J., 29: 38–38. [Taylor &; Francis Online], [Web of Science ®] , [Google Scholar] which showed that the antibodies produced by hamsters in response to a hardjo bacterin protects them against balcanica infection. Their results complement those from a passive hamster protection test conducted by ourselves. In this test 10 hamsters were each injected intraperitoneally with 0.5 ml of bovine anti-Pithardjo serum. This serum had been collected from a heifer which had received a 2 ml dose of a commercial hardjo/pomona bacterin* * Leptavoid, Wellcome (N.Z.) Ltd. eight weeks previously and had a titre of 1:96 to hardjo. Twenty-four hours later these 10 hamsters were challenged: 5 received an intraperitoneal inoculation of approximately 2 x 107 bakanica organisms and 5 received a similar inoculum of hardjo organisms. At the same time, 2 control groups of 5 hamsters were challenged with balcanica and hardjo by the same route. Leptospires could not be isolated from any of the challenged hamsters which received anti-hardjo serum when their kidneys were cultured 18 days post-inoculation. However, leptospires were isolated from the kidneys of 5 out of 5 control hamsters inoculated with hardjo and 4 out of 5 control hamsters inoculated with balcanica. An analogous trial using bovine anti-balcanica serum, collected from cattle experimentally infected with balcanica, gave almost identical results to the above trial.  相似文献   

16.
OBJECTIVE: To evaluate the ability of commercially available Escherichia coli J5 bacterin to protect rabbits from experimental challenge with Pasteurella multocida. ANIMALS: 40 P multocida-free New Zealand White rabbits. PROCEDURES: Rabbits were assigned to 1 of 4 groups of 10 rabbits each. Three of the groups were inoculated SC with J5 bacterin at 8 weeks old. Inoculation was repeated 3 and 6 weeks later. The fourth group was not inoculated and served as controls. Groups 1, 2, and 3 were given 10(9), 10(8), and 10(7) colony forming units (CFU), respectively. Response was monitored by titer assessment, using an E coli J5 antigen capture ELISA. Five weeks after the last inoculation, all rabbits were challenged with P multocida and observed for an additional 5 weeks. Clinical, hematologic, serologic, culture, and necropsy data were collected. RESULTS: Inoculation of rabbits with 10(9) CFU of E coli J5 bacterin-induced titers that were significantly greater than titers of rabbits vaccinated with 10(8) or 10(7) CFU or those in controls. The incidence of acute bacteremia was lower in rabbits with high titers. At necropsy, prevalence of lesions typical of P multocida was not significantly different among groups. Prevalence of histologic lesions was also not significantly different among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Although the bacterin induced considerable antibody response and possibly reduced the rate of bacteremia, antibodies were not protective against long-term colonization or infection of the frontal sinuses or tympanic bullae by the challenge strain of P multocida. This bacterin in its currently available form is unlikely to aid in reducing the prevalence of pasteurellosis in rabbits.  相似文献   

17.
Summary The objective of the study was to evaluate the performance of different combinations of sample type, transport medium and culture methods for the recovery of Campylobacter jejuni and C. coli from broiler flocks at primary production. Boot swabs moistened with one of four different transport media [maximum recovery diluent (n = 120), Exeter broth (EX) (n = 120), buffered peptone water (n = 120) and modified semi‐solid Cary‐Blair (n = 120)], caecal samples (n = 40) and faecal samples (n = 120) from 40 broiler flocks were compared and sensitivity estimates obtained using a Bayesian model. Samples were cultured onto mCCDA before and after enrichment in EX and incubated microaerobically at 41.5°C. Campylobacter suspect colonies were identified to the species level by multiplex PCR. Results from the Bayesian model indicated that boot swabs after enrichment had higher sensitivity (90–94%) than caecal contents before or after enrichment (84% and 89%, respectively) and faecal samples after enrichment (82%) for the detection of Campylobacter spp., although these differences were not statistically significant. Enrichment significantly increased the sensitivity of boot swab and caecal samples for detection of Campylobacter spp. and C. jejuni, respectively. However, the enrichment of caecal samples resulted in a significant decrease in the sensitivity of these samples for detection of C. coli. There was much greater variation in the sensitivity estimates of the methods for detecting C. coli than for C. jejuni, and the ranking of methods was different between the two species. Boot swabs gave the best sensitivity values for detection of C. jejuni, and enrichment culture of faecal samples was the most sensitive method for detection of C. coli.  相似文献   

18.
Abstract

In April 2011, 40% mortality of Largemouth Bass Micropterus salmoides juveniles occurred at a farm of Zhongshan City, Guangdong Province, China. Infected fish became lethargic, exhibited corkscrew and irregular swimming, and developed a distended abdomen and crooked body. Fish began to die within 2 d after the appearance of clinical signs. In order to analyze the pathogeny and diagnose the disease earlier, observation of clinical signs, cell infection, titer calculation, electron microscopy, immersion infection assay for fish, and nucleotide sequence analysis were carried out. Fathead minnow (FHM) cell cultures, inoculated with filtrate of liver and spleen homogenates from the diseased fish, developed the obvious cytopathic effect 46 h after inoculation in the primary culture and 24 h at the first passage. Typical rhabdovirus particles, 115–143 nm in length and 62–78 nm in diameter, were observed in infected FHM cells by direct transmission electron microscopy. The isolated virus produced a titer of 107.15 TCID50/mL. Immersion-Fish infected with the virus had similar clinical signs and 80% mortality with 102.5 LD50/mL. The data indicated that the rhabdovirus was the lethal pathogeny of the current disease. Based on nucleoprotein-gene nucleotide sequence multiple alignment analysis, the newly isolated virus is a strain of Siniperca chuatsi rhabdovirus (SCRV) under family Rhabdoviridae, which was initially isolated from Mandarin Fish Siniperca chuatsi. Up to the present, at least four virus strains have been isolated from diseased Largemouth Bass, which have had different clinical signs. Comparison of the clinical signs can help in an early diagnosis of the disease.

Received October 30, 2012; accepted April 19, 2013  相似文献   

19.
An investigation was made into the prevalence of leptospiral infection in cattle. An area 50 km radius was selected in a region where leptospirosis was reputedly common. Farmers volunteered 250 herds with 39 500 cows for testing and 7 500 animals were selected and sampled. Twenty-nine cows (0.4%) on 14 (5.6%) of the farms had leptospiruria at the first examination. Leptospirae were cultured from the urines of nine of these animals and all were Leptospira interrogans serovar hardjo. Serologically 12.5% of cows had titres of 1:200 or greater to hardjo and 3.5% titres of 1:200 or greater to pomona. In the Spring of 1977, there was evidence of clinical leptospirosis in calves associated with only one of the herds and no clinical leptospirosis in the 250 lactating herds, although leptospiral titres were found in 88% of them. This indicated that clinical disease was much less common than infection. We concluded that leptospirosis was of minor economic importance in dairy cattle, although it could be significant in individual herds, and a health hazard to farm workers.  相似文献   

20.
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   

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