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1.
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
2.
Shedding and Seroprevalence of Pathogenic Leptospira spp. in Sheep and Cattle at a New Zealand Abattoir 
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下载免费PDF全文 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
3.
Truong LQ Kim JT Yoon BI Her M Jung SC Hahn TW 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(12):1597-1601
Brucella infections in wildlife originate either from contact with infected livestock or from a natural sustainable reservoir in wildlife populations. As South Korea has set a goal of brucellosis eradication by 2013, it is necessary to determine the prevalence of Brucella in wildlife and wild rodents. This information will play an important role in the control of brucellosis. Because of the absence of prominent clinical signs, direct and indirect laboratory tests are essential for diagnosing brucellosis. In this study, tissue and blood samples were taken from wild animals, abandoned dogs, a cat and wild rodents, and they were tested for Brucella or Brucella-specific antibodies by isolation, PCR and serology. Results showed that 18.6% (33/177) of blood samples were positive by PCR, and 5.7% (11/194) were positive by C-ELISA. However, none of these samples yielded culturable bacteria. Of the tissue samples, 9.7% (8/82) were positive by PCR. Brucella was isolated from only one tissue culture from a Chinese water deer carcass. This Brucella species was identified as Brucella abortus biovar 1 by biotyping, 16S rRNA PCR and the Bruce-ladder PCR assay. In this study, we reported the prevalence of Brucella in wildlife, dogs, a cat and rodents by using serological and molecular methods, and we report the first isolation of B. abortus in wild Chinese water deer in South Korea. 相似文献
4.
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
5.
The technique of polymerase chain reaction (PCR) associated to hybridization was used to screen 123 samples collected from wild and synanthropic rodents captured in a cutaneous and visceral leishmaniasis endemic area in the State of Minas Gerais, Brazil. The detection of Leishmania spp in naturally infected rodents is of fundamental importance for incriminating them as possible reservoir hosts of the diseases in Minas Gerais. A total of 62 specimens belonging to wild (Thrichomys apereoides, Oryzomys subflavus, Galea spixii, Bolomys lasiurus and Wiedomys pyrrhorhinos) and synanthropic (R. rattus) rodent species were captured in different ecotopes. Blood and skin samples were submitted for PCR analyses followed by molecular hybridization with specific probes for the three Leishmania-species complexes. Fifteen samples were found positive after PCR-hybridization and identified as follows: nine belonging to the L. mexicana complex, three to the L. braziliensis complex and three to the L. donovani complex. Positive PCR results were found in 11 out of the 61 (18%) blood samples and in four out of the 62 (6.4%) skin fragments screened. R. rattus and T. apereoides were the most abundant species in the area also presenting high prevalence of natural infection. The presence of parasite DNA belonging to L. braziliensis, L. mexicana and L. donovani complexes was confirmed in several individuals of a rodent species, R. rattus. This work is the first report of the detection of L. (L.) chagasi in a naturally infected T. apereoides. The utility of filter paper as a substrate for PCR analyses and the efficacy of the procedure associated to the hybridization is emphasized. 相似文献
6.
Rats are known to be the most important reservoirs of Leptospira spp. However, the leptospiral dose and age at which rats become resistant to Leptospira infection are not yet well elucidated. Aimed to characterize leptospirosis in rat pups, we found that suckling pups (4-, 7-, and 14-day old) are susceptible to leptospires and resistance starts from the weaning age (23-day old). Susceptibility of rat pups was also affected by the infecting dose of the organisms. Jaundice, decrease in body weight, and neurological symptoms prior to moribundity was evident in infected suckling pups. However, 23-day-old infected pups did not manifest any pathological changes and were able to survive the infection similar to adult rats. Based on these results, we propose the suckling rat pup as a novel animal model of human leptospirosis to investigate pathogenesis, development of host resistance, and the mechanisms involved in rats becoming maintenance hosts for leptospires. 相似文献
7.
Serum samples were examined for evidence of leptospiral agglutinins from 928 sheep from 45 lines and kidneys from 12 of these lines for evidence of leptospiral infection. All sheep had been submitted for slaughter at meat works in the Manawatu. Serological results were analysed using the results at a minimum serum dilution in the microscopic agglutination test (MAT) of 1:24 and at a minimum dilution of 1:48. It was shown that a minimum dilution of 1:24 resulted in many non-specific or cross-reactions. A minimum dilution of 1:48 was more accurate for detecting the serological prevalence of specific agglutinins to leptospires in ovine sera. Twenty percent of the sheep had titres of 1:48 or greater to hardjo, 3.8% to pomona, 2.6% to tarassovi, 2.3% to copenhageni and 2.7% to ballum. No titres of 1:48 or greater to australis were detected. Serovar hardjo was isolated from the kidneys of three animals in one line. Eighteen months later 291 serum samples and 95 urine samples were collected from live animals on the property from which the three hardjo infected animals originated. No titres to hardjo were detected in the sera of lambs, but a serological prevalence of 44% and 84% to this serovar was demonstrated in the hoggets and ewes respectively. No leptospires were demonstrated in any of the urine samples. These results show that sporadic infection of sheep with hardjo can occur but they also indicate that infection with this serovar is not endemic and that sheep are unlikely to act as maintenance hosts for hardjo in New Zealand. 相似文献
8.
9.
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
10.
AIM: To investigate the role of free-living animals such as sparrows, rodents and flies as potential reservoirs of Campylobacter spp on a dairy farm, and to assess the genetic diversity among Campylobacter isolates from the farm and an urban source. METHODS: A total of 290 samples (bovine, passerine and rodent faeces, and whole flies) were collected from a large commercial dairy farm in the Manawatu district in New Zealand, and from faeces from urban sparrows in a nearby city. Other samples collected from the dairy farm included five from silage, two from aprons worn by workers during milking, two from workers' boots and two from water in troughs in a paddock. Isolates of thermophilic Campylobacter spp were identified morphologically and phenotypically and further characterised molecularly using pulsed-field gel electrophoresis (PFGE) and the restriction enzyme SmaI. RESULTS: Campylobacter jejuni was the only Campylobacter species isolated from all samples. The highest prevalence was found in faeces from dairy cows (54%), followed by faeces from sparrows from the urban area (40%) and the farm (38%), and from rodents (11%) and whole flies (9%). Other samples from the farm environment such as silage, trough water, and workers' aprons and boots were also positive for C. jejuni. Of the 22 restriction patterns obtained, seven were common to more than one source. CONCLUSIONS: Cattle, sparrows, rodents and flies are potential reservoirs of C. jejuni on dairy farms. Identical clones of C. jejuni carried by cattle, sparrows, flies and rodents probably indicate a common source of infection. The high level of asymptomatic carriage of C. jejuni by healthy dairy cows could be sufficient to maintain infections within the dairy farm surroundings via environmental contamination. 相似文献
11.
Echinococcus multilocularis causes a rare but potentially lethal zoonotic infection in humans. This tapeworm is known to be endemic in foxes in several countries of Western and Central Europe. In Western Europe, the common vole (Microtus arvalis) and the water vole (Arvicola terrestris) are considered to be the most important intermediate host species of this cestode whereas the red fox is by far the most important final host. The purpose of this study was to provide data on the prevalences in Wallonia (Southern part of Belgium) both in the red fox and in different potential intermediate hosts. A total of 990 red foxes were examined between January 2003 and December 2004 for the presence of E. multilocularis. The average prevalence was 24.55% (22.38-27.87). Out of 1249 rodents or insectivores belonging to the species Apodemus sylvaticus, Arvicola terrestris, Clethrionomys glareolus, Microtus arvalis, Microtus agrestris and Sorex araneus, only one M. arvalis (out of 914-0.11% (0.003-0.61) and one C. glareolus (out of 23-4.3% (0.1-21.9) were found to be infected. However, the muskrat (Ondatra zibethicus) seems to be a good intermediate host as 11.18% (9.72-12.76) of the animals (n=1718) were found to be infected. A positive correlation was found between the prevalences in foxes and in muskrats in each of the different geological regions. This study indicates that the muskrat is highly sensitive to this zoonotic tapeworm and could perhaps represent a good bioindicator when studying the epidemiology of this parasitic infection in Belgium and in other countries where the muskrat is present. 相似文献
12.
A. E. Buchholz A. R. Katz R. Galloway R. A. Stoddard S. M. Goldstein 《Zoonoses and public health》2016,63(8):584-587
Leptospirosis is considered the most widespread of zoonotic diseases. It was a notifiable disease in the United States until 1995 and was reinstated to the list of nationally notifiable diseases in 2014. During the time of national surveillance, Hawaii consistently led the nation in reported annual incidence rates. Leptospirosis has remained a reportable disease in Hawaii. Significant changes have been documented since the early 1970s in the predominant serogroup infecting humans in Hawaii: infections due to Icterohaemorrhagiae have declined while infections due to Australis have increased. A recent study from Hawaii demonstrated that Australis was an uncommon infecting serogroup for small mammal hosts. Swine have not been previously studied in Hawaii but are well‐recognized maintenance hosts for leptospires belonging to the Australis serogroup. This study was undertaken to assess the prevalence of Leptospira antibody in feral swine in Hawaii. From January 2007 through December 2009, blood samples were collected opportunistically from feral swine. Using the microscopic agglutination test, we found antibody titres ≥1 : 100 to leptospires in 272 (33.8%) of 804 feral swine. The most frequently reacting serovars to the swine sera were Icterohaemorrhagiae (Icterohaemorrhagiae serogroup) (41.5%) and Bratislava (Australis serogroup) (33.8%). The high seroprevalence and presumptively infecting serovars suggest a link between swine and human infection. 相似文献
13.
The isolation of leptospires from buffaloes worldwide is still limited to a few strains. Thus, the aim of this study was to describe the first Leptospira isolate from buffalo urine, assigned to the Sejroe serogroup, which does not belong to the Wolffi subgroup, traditionally isolated in Brazil. A total of 244 urine samples of water buffaloes (Bubalus bubalis) raised in the Brazilian Amazon were subjected to bacteriological culturing and polymerase chain reaction (PCR) for the detection of leptospires. The obtained isolate was characterized by serogrouping using polyclonal antibodies, partial DNA sequencing, Hardjo-Bovis-specific PCR, multiple-locus variable-number tandem repeat analysis (MLVA/VNTR) and experimental infection in hamsters. PCR was performed on the urine samples; 11/244 were positive (4.5 %) for Leptospira, and only one isolate was recovered (0.4 %). Regarding characterization, the isolate was assigned to the Sejroe serogroup with high titers (12,800) for the Saxkoebing and Sejroe serovar antisera. The isolate was negative for Hardjo-Bovis-specific PCR, and the species Leptospira borgpetersenii was identified by DNA sequencing. The MLVA results showed that the VNTR profile of the isolate was 1−2-5, compatible with that of serovars Sejroe/Istrica. In the experimental infection in hamsters, the animals did not develop clinical signs, and no macroscopic lesions were observed on the organs at necropsy; however, the strain was detected in the kidneys, uterus, and testicles of the animals. The isolate described herein highlights infection by Sejroe strains that may be overlooked in buffaloes and that may be different from those normally isolated and used in serological studies. 相似文献
14.
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
15.
Introduction and purposeTularemia is a zoonotic disease, the most important hosts of which are rodents. Endemic regions and reservoirs of F. tularensis are not well-researched areas in Iran. The present study aimed to study F. tularensis infection in the rodent populations of western Iran.Materials and methodsSamples were collected in different areas of Kabudar Ahang County in Hamadan province (west of Iran) from 2014 to 2017. Tularemia serological and molecular tests were conducted using the tube agglutination test and Real-time PCR method tracking the ISFtu2 gene. Positive serum samples were evaluated for cross-reactivity with brucellosis.ResultsA total of 433 rodents, collected from 33 localities, were included in the study. The most abundant species belonged to the Persian jird (Meriones persicus; 75.5%), and Libyan jird (Meriones libycus; 10.1%). Among the studied samples, three (0.74 %) were seropositive and five (1.15%) were PCR positive. Seropositive samples were two M. persicus and one M. libycus, and PCR positive rodents were four M. persicus and one M. vinogradovi. Tularemia seropositive samples showed no cross-reactivity with brucellosis.ConclusionGiven the presence of infection in rodents with tularemia agent in the studied area, it is crucial to elucidate the risks of rodent exposure to tularemia for physicians, health personnel and the general population. 相似文献
16.
AIM: To investigate the role of free-living animals such as spar- rows, rodents and flies as potential reservoirs of Campylobacter spp on a dairy farm, and to assess the genetic diversity among Campylobacter isolates from the farm and an urban source. METHODS: A total of 290 samples (bovine, passerine and ro- dent faeces, and whole flies) were collected from a large com- mercial dairy farm in the Manawatu district in New Zealand, and from faeces from urban sparrows in a nearby city. Other samples collected from the dairy farm included five from silage, two from aprons worn by workers during milking, two from workers' boots and two from water in troughs in a paddock. Isolates of thermophilic Campylobacter spp were identified mor- phologically and phenotypically and further characterised mo- lecularly using pulsed-field gel electrophoresis (PFGE) and the restriction enzyme SmaI. RESULTS: Campylobacter jejuni was the only Campylobacter species isolated from all samples. The highest prevalence was found in faeces from dairy cows (54%), followed by faeces from sparrows from the urban area (40%) and the farm (38%), and from rodents (11%) and whole flies (9%). Other samples from the farm environment such as silage, trough water, and work- ers' aprons and boots were also positive for C. jejuni. Of the 22 restriction patterns obtained, seven were common to more than one source. CONCLUSIONS: Cattle, sparrows, rodents and flies are po- tential reservoirs of C. jejuni on dairy farms. Identical clones of C. jejuni carried by cattle, sparrows, flies and rodents prob- ably indicate a common source of infection. The high level of asymptomatic carriage of C. jejuni by healthy dairy cows could be sufficient to maintain infections within the dairy farm sur- roundings via environmental contamination. 相似文献
17.
Karen E. Shearer Michael J. Harte Davor Ojkic Josepha DeLay Douglas Campbell 《The Canadian veterinary journal. La revue veterinaire canadienne》2014,55(3):240-248
A total of 460 kidney samples from wildlife (beavers, coyotes, deer, foxes, opossums, otters, raccoons, skunks) were obtained from road-kill and hunter/trapper donations in Ontario between January 2010 and November 2012. The objectives of the study were to detect Leptospira spp. by immunohistochemistry and polymerase chain reaction (PCR), to map presence of leptospires in wildlife relative to livestock and human populations, and to characterize positive samples by sequencing and comparison to leptospires known to affect domestic animals and humans. The proportion of samples that tested positive ranged from 0% to 42%, with the highest rates in skunks and raccoons. Leptospira spp. were present in kidneys of wildlife across Ontario, particularly in areas of high human density, and areas in which livestock populations are abundant. The PCR was too weak in most samples to permit genotyping and examination of the relationship between the leptospires found in this study and those affecting domestic animals and humans. 相似文献
18.
Umhang G Raton V Comte S Hormaz V Boucher JM Combes B Boué F 《Veterinary parasitology》2012,188(3-4):301-305
The life cycle of the zoonotic parasite Echinococcus multilocularis is predominantly sylvatic, involving foxes as definitive hosts infected by predation of rodents, the intermediate hosts. The North-Eastern French departments of Meuse and Haute-Sa?ne are highly endemic, with an estimated fox prevalence of 41% and 36% respectively. Although most of the parasites' biomass occurs in foxes, domestic dogs can also be infected, leading to a major risk of human infection due to the close proximity of dogs and owners. In the present study, dog faeces were collected after praziquantel treatment provided by veterinarians. In all, 860 faecal samples were collected throughout Meuse (n=493) and Haute-Sa?ne (n=367). Intestinal helminth eggs were isolated from the faeces using a flotation technique and observed by microscopy. Parasite species were identified in samples positive for taeniid eggs by sequence analysis after PCR amplification. To study the factors associated with infestation, each sample was linked to a questionnaire filled in by the dog owners. Taeniid eggs were observed in seven faecal samples (0.8%) but none of them were positive for E. multilocularis. Thus, the apparent prevalence of E. multilocularis in dog populations is lower than 1.00% for Haute-Sa?ne and lower than 0.75% for Meuse. In Haute-Sa?ne, a high proportion of dogs observed suspected preying on rodents were not dewormed monthly. In endemic areas, these dogs must be considered at risk of transmitting E. multilocularis to humans. 相似文献
19.
The prevalence of the fox tapeworm in foxes (final host) and muskrats (one of the intermediate hosts) in the Netherlands and Europe has been discussed. The tapeworm was found in 9.4% of the investigated foxes from the province of Groningen and in 0.2% of the muskrats from the same region. Also in the province of Limburg positive foxes were found, but no positive muskrats. Possible ways of infection for humans are described together with methods for prevention. It is concluded that at this moment risks for humans to become infected are minimal, but vigilance and monitoring of foxes and muskrats remains needed. 相似文献
20.
A Hossain Farid 《Acta veterinaria Scandinavica》2013,55(1):10