Evidence for bovine besnoitiosis being endemic in Italy--first in vitro isolation of Besnoitia besnoiti from cattle born in Italy

Until 2009, bovine besnoitiosis had never been considered endemic in Italy and the only report on the disease in this country referred to animals imported from France shortly before. However, recently, an autochthonous outbreak of bovine besnoitiosis was reported in four herds located at the intersection of the borders between Emilia-Romagna, Toscana and Marche (Northern Apennine Mountains), which has led to an increased awareness concerning this disease. The present study describes a further outbreak of bovine besnoitiosis in Italy. The afflicted herd was a dairy herd with no evidence for contact with cattle from regions known to be endemic for bovine besnoitiosis. The farm investigation was initiated after a three-year old Holstein Friesian dairy cow with generalized thickening and lichenification of the skin was diagnosed with bovine besnoitiosis. The clinical diagnosis was confirmed by gross pathology, histopathology, serology and PCR. Bradyzoites released from tissue cysts obtained from the skin of this animal enabled the first in vitro isolation of Besnoitia besnoiti in Italy. This isolate was named Bb-Italy1. Sequencing of a 2118 bp spanning region including the complete internal transcribed spacer 1 and parts of the 18S and the 5.8S rRNA gene from DNA extracted from skin-derived zoites revealed a 99.9% identity to sequences known for other B. besnoiti isolated from cattle in Europe. Two GKO mice which had been inoculated intraperitoneally with bovine skin-derived bradyzoites became ill 7 days post inoculation. Parasitophorous vacuoles with multiplying zoites were observed in the cell culture inoculated with peritoneal fluids of these mice and a B. besnoiti infection in the mice and in the cell culture could be confirmed by real-time PCR. A serological investigation in the afflicted herd using immunoblots and an immunofluorescent antibody test (IFAT) revealed an overall herd seroprevalence of 9.7% (31/321), whereas within the female animals older than 2 years 17.0% (29/171) of the dams were tested positive. With one exception, an imported cow from Germany, all the seropositive animals were born in Italy. In connection with previously described autochthonous cases of bovine besnoitiosis the case described herein suggests that bovine besnoitiosis should be considered endemic in Italy.     

A modified agglutination test for the diagnosis of Besnoitia besnoiti infection

Bovine besnoitiosis is caused by Besnoitia besnoiti, an obligate intracellular apicomplexan parasite. Affected animals present cutaneous and systemic manifestations and the disease may lead to considerable economic losses. Although generally associated to tropical and subtropical areas, bovine besnoitiosis is now considered an emergent disease in Europe, due to the increasing number of new cases and apparent geographical expansion. In this study we evaluated the performance of a modified agglutination test (B-MAT) in the serodiagnosis of bovine besnoitiosis in comparison to the indirect immunofluorescent-antibody test (IFAT). To establish optimal protocol conditions we used bovine sera with a known infection status for B. besnoiti infection. Positive animals (n=36) presented B. besnoiti dermal cysts and anti-B. besnoiti specific antibodies, as determined by the indirect immunofluorescence test (IFAT). Negative animals (n=103) were from non-endemic areas in Portugal and negative by the IFAT. From here, we evaluated the sensitivity and specificity of the B-MAT relative to the IFAT with a panel of sera from herds with history of bovine besnoitiosis in Portugal, Spain and France (n=402), using three serum dilutions (1:80, 1:160, 1:320). Considering the positive cut-off at 1:160 serum dilution, the B-MAT showed an almost perfect test agreement with the IFAT; (κ=0.968; 95% CI: 0.941-0.996) with a relative sensitivity of 97.2% (95% CI: 94.1-100%) and a relative specificity of 99.3% (95% CI: 98.4-100%). As a simple and inexpensive technique the B-MAT represents a valuable tool for the diagnosis and study of the epidemiology of bovine besnoitiosis.     

Quantitative real time polymerase chain reaction assays for the sensitive detection of Besnoitia besnoiti infection in cattle

Bovine besnoitiosis, an economically important disease in cattle in some countries of Africa and Asia, is emerging in Europe. The definitive host of Besnoitia besnoiti, the causative agent of bovine besnoitiosis, is unknown and the transmission of the parasite is not completely understood. Sensitive and quantitative DNA detection methods are needed to determine whether serologically positive animals are infectious and to examine the role of vectors (e.g. haematophagous insects) in the transmission of the parasite. To this end, we established two different 5'-nuclease quantitative assays to detect B. besnoiti infection in cattle and to estimate the parasite load in samples (BbRT1 and BbRT2). These PCRs are based on the sequence of the internal transcribed spacer region 1 (ITS-1) of the ribosomal RNA gene. Tests with serial dilutions of B. besnoiti genomic DNA in a buffer containing 100 ng/μl bovine DNA revealed a detection limit of 0.01 pg genomic B. besnoiti DNA. Reliable quantification was possible in samples containing ≥1 pg B. besnoiti genomic DNA with a coefficient of variation of ≤ 2%. To estimate the diagnostic sensitivity of the tests, skin biopsies and scrapings from the mucous membrane of the vestibulum vaginae (vaginal scrapings) were taken from cattle with clinical signs of chronic besnoitiosis. Regardless of the real time PCR assay used, 90.7% (39/43) of these animals were positive in at least one of two samples (skin or vaginal scrapings). Antibody titers, as determined by an immunofluorescent antibody test, and the threshold cycle values of the real time PCR obtained for skin samples and vaginal scrapings, were significantly correlated. The specificity of the PCRs was confirmed using genomic DNA from related parasites, including genomic DNA of Besnoitia spp., Neospora caninum, Toxoplasma gondii, Hammondia hammondi, Hammondia heydorni, Isospora spp., Sarcocystis spp., Eimeria bovis, Cryptosporidium parvum, and Trypanosoma brucei brucei. Since the sequence of the ITS-1 region of B. besnoiti is identical with that of Besnoitia species isolated from donkeys (Besnoitia bennetti), and reindeer (Besnoitia tarandi), both real time PCRs detected also DNA of these parasites. One of the B. besnoiti real time PCRs, BbRT1, but not BbRT2, cross-reacted with Besnoitia darlingi, Besnoitia oryctofelisi, and Besnoitia neotomofelis when large amounts of genomic DNA (10 ng) were used. The other B. besnoiti real time PCR assay (BbRT2) was specific for B. besnoiti, B. bennetti and B. tarandi, but did not react when 10 ng DNA of other related parasite species from the genus Besnoitia or other genera were subjected to analysis.     

Immunodiagnosis of Besnoitia besnoiti infection by ELISA and Western blot

Besnoitia besnoiti, an obligate intracellular apicomplexan protozoan parasite, is the causative agent of bovine besnoitiosis. This infection may dramatically affect body condition and lead to irreversible infertility in males, resulting in important economical losses in livestock production. Identification of serologically positive animals is of major relevance to elaborate appropriate measures of control. While identification of clinical cases is relatively easy to carry out, the finding of subclinical forms of infection is more difficult, thus serology is considered as an appropriate diagnostic tool. In view to improve and validate immunodiagnosis, we evaluated an enzyme-linked immunosorbent assay (ELISA), complemented with a Western blot (both using a somatic B. besnoiti-tachyzoite antigen) to detect anti-B. besnoiti antibodies in bovine sera. The comparative evaluation of the 2 methods, using 13 sera from animals affected by the chronic phase of besnoitiosis and 10 asymptomatic carriers, yielded a diagnostic sensitivity of 87% for ELISA and 91% for Western blot analyses. Specificity was tested with sera from animals with confirmed Toxoplasma gondii (n=5) and Neospora caninum (n=12) infection, and with 64 negative sera from either an endemic or a non-endemic area. The ELISA specificity ranged between 96.4% and 98%, the Western blot specificity between 96.4% and 100%. The present study demonstrated that ELISA and Western blot, using in vitro generated somatic B. besnoiti antigen, is a useful tool combination to reliably detect animals that have been exposed to B. besnoiti infection, including both asymptomatic and symptomatic courses of disease.     

Isolation of Besnoitia besnoiti from infected cattle in Portugal

Besnoitia besnoiti, an obligate intracellular protozoan parasite belonging to the phylum apicomplexa, is the causative agent of bovine besnoitiosis. Besnoitiosis is responsible for significant losses in the cattle industry of Africa and Mediterranean countries due to the high morbidity rate, abortion and infertility in males. The acute stage of disease is associated with the proliferative forms (tachyzoites) and is characterized by fever, whimpery, general weakness and swelling of the superficial lymph nodes. During the following chronic stage, a huge number of cysts are formed mainly in the subcutaneous tissues. This process is non-reversible, and chronic besnoitiosis is characterized by hyper-sclerodermia, hyperkeratosis, alopecia and, in bulls, atrophy, sclerosis and focal necrosis that cause irreversible lesions in the testis. In this paper we report on the identification of large cysts in the skin of a cow and a bull in Portugal, which presented loss of hair and enlargement and pachydermis all over the body. The observation of a two-layered cyst wall within the host cell, the encapsulation of the host cell by a large outer cyst wall, and the subcutaneous localization of the cysts within the host, were characteristic for B. besnoiti. The parasites were isolated from the infected animals and successfully propagated in Vero cells without prior passages in laboratory animals. Morphological characterization of B. besnoiti tachyzoites and the amplification of the 149 bp segment from the internal transcribed spacer 1 (ITS1), aided with specific primers, confirmed the identification of B. besnoiti.     

Evaluation of a commercial ELISA for the specific detection of antibodies against Besnoitia besnoiti

Bovine besnoitiosis is an economically important disease in cattle caused by the protozoan parasite Besnoitia besnoiti, which occurs endemically in many countries of Africa and Asia and is spreading in Europe. Serological identification of subclinically infected cattle is important to avoid the introduction of infected animals into naive herds. Here we determine the sensitivity and specificity of the PrioCHECK^® Besnoitia Ab, a serological test recently introduced into the European market. Analytical specificity was examined using sera from animals experimentally infected with parasites related to B. besnoiti (n = 27). Three animals experimentally infected with Neospora caninum or Toxoplasma gondii showed inconclusive reactions in the ELISA (percent positivity relative to the positive control [PP] 10% ≤ 20%) while all other sera reacted negative (PP < 10%). An estimate of the diagnostic specificity was obtained by analysing field sera from bovine herds without besnoitiosis but with abortion problems associated to N. caninum (n = 403). The analysis revealed a specificity of 94.3% or 96.8% depending on the applied cut-off (PP 10% or 20%, respectively). Sensitivity was assessed with sera from 110 animals of a herd in Germany where clinical bovine besnoitiosis was first diagnosed in September 2008. A positive serological reference standard was defined regarding sera from animals as reference positive, if these animals had tested positive in at least two of a panel of three other serological tests (two different B. besnoiti immunoblots and one immunofluorescence antibody test) on both of two sampling dates, November 2008 and April 2009. A diagnostic sensitivity of 91.8% or 75.5% was determined for sera collected in November 2008 and a sensitivity of 82.7% or 50% for sera collected in April 2009 (cut-off PP 10% or PP 20%, respectively). The marked drop in sensitivity from November 2008 to April 2009 was predominantly observed in reference-positive cattle without clinical signs. We conclude that PrioCHECK^® Besnoitia Ab is a valuable diagnostic tool to detect clinically infected animals. Thus it may be used to support control measures, e.g., for the separation of infected animals from the remaining herd to avoid a further transmission of the infection within the herd.     

Exploring the life cycle of Besnoitia besnoiti - experimental infection of putative definitive and intermediate host species

The biology of Besnoitia besnoiti, the cause of bovine besnoitiosis, is poorly understood. Its definitive host is unknown, and information on potential intermediate hosts is scarce. In order to investigate potential definitive and intermediate hosts for European isolates of B. besnoiti, domestic dogs, cats, rabbits, guinea pigs (Cavia porcellus), gerbils (Meriones unguiculatus), common voles (Microtus arvalis) and NMRI-mice were inoculated with B. besnoiti isolated from naturally infected German cattle. Dogs and cats were fed 5×10(6)B. besnoiti tachyzoites (isolate Bb-GER1), or tissue cysts containing at least 2×10(7)B. besnoiti bradyzoites obtained from the skin of a naturally infected Limousin cow from the same herd where strain Bb-GER1 was isolated. Rodents and rabbits were subcutaneously inoculated with either 5×10(5) Bb-GER1 tachyzoites or 5×10(5) bradyzoites. Groups of 2-4 non-inoculated animals of each species were monitored as negative controls. Feces from all dogs and cats were daily examined by a sedimentation-flotation technique for at least 11 weeks after inoculation but no B. besnoiti oocysts were identified. Cats fed tachyzoites and dogs did not seroconvert, but specific antibodies to B. besnoiti tachyzoites were detected by IFAT (titer≥100) in 2 out of 3 cats fed tissue cysts since 5-7 weeks post infection. By immunoblot, these two cats exhibited a reaction pattern against tachyzoite antigens similar to that observed in naturally infected cattle. Antibodies against B. besnoiti tachyzoites were detected in all inoculated rodent species and rabbits by both, IFAT and immunoblot since 3 weeks post-inoculation. Rabbits and rodents, subcutaneously inoculated with same doses of inactivated bradyzoites remained serologically negative (IFAT titer<50). Clinical signs observed in the inoculated rabbits included fever, serous conjunctivitis and transient swelling of the testes. No clinical abnormalities were noticed in the other tested animal species. Voles developed pneumonia as observed by histological examination. B. besnoiti-DNA was detected by PCR in blood from rabbits, gerbils and voles at 9 days post-infection, and in skin, heart, lung, striated muscle and kidney tissues from voles at 19-21 weeks post-infection. Domestic dogs and cats could not be shown to be definitive hosts of B. besnoiti, but cats seroconverted after feeding on B. besnoiti tissue cysts indicating that B. besnoiti stages had invaded the cats' tissues. The molecular and serological results from this study indicate that European B. besnoiti isolates may infect cats, rabbits, guinea pigs, gerbils, mice and voles; however a persistence of the parasite could be demonstrated only in voles.     

Serodiagnosis of bovine besnoitiosis by ELISA and immunofluorescence tests

Sera from non-infected cattle and cattle infected with Anaplasma, Babesia, Theileria and Sarcocystis were tested for antibodies to Besnoitia in ELISA and immunofluorescence tests (IFT) with Besnoitia besnoiti of blue wildebeest origin as antigen. Only 2 out of 86 sera gave false positive reactions in ELISA and none in the IFT, indicating a high specificity for the tests. Three-hundred-and-three bovine sera from 3 farms in an area endemic for besnoitiosis were similarly tested and the results were correlated with clinical findings based on visual inspection for typical symptoms and the presence of cysts in the scleral conjunctiva. Most of the positive tests were observed in cattle older than 1 year. Of the cases with scleral cysts, 68,7% were positive in the ELISA and 81,74% in the IFT. However, 45,74% (ELISA) and 49,47% (IFT) of the clinically negative cattle were clinically positive, indicating a high incidence of clinically inapparent infection. These results indicate a relatively low sensitivity for these serological tests. An unexpected finding was that the ELISA remained negative for at least 60 days after experimental infection of the cattle, the maximum period for which tests were done, whereas the IFT became positive. No antibodies against B. besnoiti could be found in human sera. Besnoitia jellisoni antigen gave positive results with B. besnoiti antibodies in ELISA, but not in the IFT.     

Immunological relationship between Neospora caninum and Besnoitia besnoiti

Neospora caninum is a coccidian parasite identified as a major cause of abortion in cattle. A combined infection of N. caninum with another taxonomically related parasite of cattle, Besnoitia besnoiti can occur in geographical areas endemic for both species. Both infections are routinely diagnosed serologically, and incorrect diagnosis could occur if immunological cross-reactivity exists between the two parasites. To investigate the possible degree of cross-reactivity, we compared results obtained with two serological techniques, immunofluorescent antibody test (IFA), and Western blot analysis on known positive and negative sera. The test sera were derived from naturally infected cattle and from experimentally infected Mongolian gerbils. In IFA of bovine sera, no cross-reactvity was detected at the commonly used serum dilution cutoffs of 1:200 for N. caninum and 1:256 for B. besnoiti. However, at 1:64 dilution of both cattle and gerbil sera, anti-N. caninum sera reacted with B. besnoiti antigen in some individual samples. Anti-B. besnoiti serum did not react with N. caninum antigen at any dilution. This low level one directional cross-reactivity was confirmed by Western blot analysis. B. besnoiti antigen showed two immunoreactive bands when probed with anti-N. caninum serum, while no bands appeared when N. caninum antigen was probed with B. besnoiti antiserum. Immunization and challenge experiments in the highly susceptible Mongolian gerbil (Meriones unguiculatus) showed essentially no cross-protection between N. caninum and B. besnoiti.     

Enzyme linked immunosorbent assay for detection of antibodies againstBesnoitia besnoiti in cattle

The use of the ELISA method for the detection of antibodies to B. besnoiti in cattle is described and compared to the IFAT technique. One hundred and twenty-one sera were examined, of which 61 were sera of calves experimentally infected with B. besnoiti, 52 sera from field animals and eight were sera with high titres of antibodies to other parasites. The specificity of both assays correlates but ELISA seemed to be more sensitive. The ELISA technique provides a rapid and reliable method for the screening of B. besnoiti infection in cattle.     

Serological studies on bovine besnoitiosis in Israel

Sera from 1,700 beef and dairy cattle in Israel were tested for besnoitiosis by indirect immunofluorescence. Over 90% of dairy animals were negative whereas about 50% of beef cattle were positive. Among beef bulls the percentage of positive reactors increased with age but in beef cows the pattern was less clear; 36% of young imported bulls negative upon arrival developed antibody titres after one season on the range. Cross-tests with Toxoplasma gondii antiserum and antigen indicated that the results of the survey for besnoitiosis were unlikely to have been influenced by Toxoplasma antibodies.     

Identification and partial purification of soluble antigens from culture-grown Besnoitia besnoiti endozoites

Soluble antigens from culture-grown Besnoitia besnoiti endozoites were identified following their partial purification by affinity chromatography. A specific eluate obtained after affinity chromatography on a column to which antibodies from serum of a naturally infected cow were bound exhibited seven polypeptide bands on sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Five bands were observed in the eluate from an immunoadsorbent to which antibodies from an experimentally infected calf were coupled. Eluted antigens were reactive in the enzyme-linked immunosorbent assay (ELISA). Reactivity of electrophoretically resolved antigens with sera of endozoite-inoculated cattle and sera from field cases of besnoitiosis were studied using the Western immunoblotting technique.     

Caprine besnoitiosis: an emerging threat and its relationship to some other infections of ungulates by Besnoitia species

Caprine besnoitiosis, caused by the cyst-forming protozoal apicomplexan Besnoitia caprae appears to be endemic in Kenya, Nigeria and Iran, but has yet to be detected in other parts of the world. The infection causes an important parasitic disease of goats in affected developing countries. Bovine besnoitiosis, is a widespread disease of cattle in Africa, Asia (but not Iran) and southern Europe. Recent epidemiological data confirm that the incidence and geographical range of bovine besnoitiosis in Europe is increasing, which is why growing attention has been given to the condition during the past decade. This paper reviews pertinent information on the biology, epidemiology, pathology, clinical signs, diagnosis and control of caprine besnoitiosis, together with its similarities to, and differences from, bovine besnoitiosis. The serious economic consequences of besnoitiosis on goat breeding and local meat and hide industries is also considered.     

Trichinella spiralis infection induces β-actin co-localized with thymosin β4

A serological survey for Neospora caninum and Besnoitia besnoiti was carried out in beef and dairy cattle in South Australia. Serum samples of dairy cattle (n=133) from 9 properties and tank milk samples from a further 122 dairy herds were tested. An additional 810 sera from beef cattle from 51 properties were also tested. Testing at the individual animal level by IDEXX NEOSPORA X2 Ab test ELISA revealed a low prevalence of N. caninum antibodies of only 2.7% (95% CI; 1.6-3.7%) sera positive, as did the milk testing that showed 2.5% (95% CI; 1.4-3.6%) of tank milks being positive. At the herd level, 29.4% (95% CI; 16.9-41.9%) of beef, and 44.4% (95% CI; 12.0-76.9%) of dairy cattle herds showed serum antibodies. The highest within-herd prevalence in beef was 20% and 25%in dairy, which explains the low herd prevalence in dairy detected by bulk milk testing. Testing for B. besnoiti antibodies by PrioCHECK(?) Besnoitia Ab 2.0 ELISA initially identified 18.4% (95% CI: 15.8-21.0%) of 869 individual cattle sera as positive by ELISA at the manufacturer's suggested cut-off threshold (15 PP). Additional tests by immunoblot and IFAT, however, could not confirm any of the ELISA results. The use of a higher (40 PP) threshold in the ELISA is suggested to improve specificity. There is thus no evidence of B. besnoiti infection in South Australian cattle.     

Bluetongue serotype 8 vaccine coverage in northern and south-eastern England in 2008

A postal survey of all registered cattle and sheep farmers in East Anglia was carried out from July 2008 to determine bluetongue virus serotype 8 (BTV-8) vaccine uptake in the region. The vaccine was available to farmers in this region from May 2008. The survey was repeated in Cumbria and Northumberland at the beginning of 2009. In these regions, the vaccine was not available until September 1, 2008. Holding-level vaccine uptake was estimated to be 85 per cent (95 per cent confidence interval [CI] 83 to 87 per cent, n=1623) in East Anglia and 36 per cent (95 per cent CI 32 to 40 per cent, n=633) in northern England. A telephone follow-up of non-responders reduced these estimates to 79 and 29 per cent in East Anglia and northern England, respectively. In both regions, vaccine coverage was higher in sheep than in cattle, with 92 per cent of sheep in East Anglia having been vaccinated. The proportion of holdings that had applied the vaccine or were intending to apply the vaccine in 2009 in the northern region was 51 per cent (95 per cent CI 47 to 54 per cent, n=664), with a further 37 per cent undecided at the time of response.     

Specific antibodies to Besnoitia besnoiti precipitated from serum of cattle by live parasites and by soluble antigen

Specific precipitable antibodies of both IgG and IgM classes were detected in sera of cattle naturally infected with B. besnoiti. The amount of specific antibodies of the IgG class precipitated by soluble antigen was in the range of 17-50 micrograms/ml serum while that of the IgM class ranged between 5 and 24 micrograms/ml serum. Specific antibodies precipitated by live B. besnoiti parasites were in amounts of 10 to 22 micrograms/ml serum for IgG and 4 to 26 micrograms/ml serum for IgM. Different ratios of IgG/IgM were obtained by the two methods of precipitation. This might indicate that antibodies to B. besnoiti of the IgM class can be precipitated and detected in sera of naturally infected cattle in similar amounts either by live parasites or by solubilized antigen, whereas antibodies of the IgG class can be preferentially detected when solubilized antigen is used for precipitation.     

A simple field diagnostic smear test for bovine besnoitiosis

A fast and inexpensive skin biopsy smear was used for confirming suspected clinical cases of bovine besnoitiosis. The technique was based on the demonstration of Besnoitia besnoiti bradyzoites (cystic stages), which appeared stumpy, each organism 6.2 microns by 3.1 microns in size, or banana-shaped, 7.7 microns by 1.5 micron in affected skin smears. A more rapid non-surgical technique, scleral conjunctival scraping, revealed similar bradyzoites, thus enhancing the diagnostic value of conjunctival cysts in more chronic infections. The technique will aid prompt management decisions to contain suspected outbreaks in herds not routinely served by tissue-processor-equipped diagnostic laboratories.     

Preliminary findings from an experimental study of caprine besnoitiosis in Kenya

Inoculation of cystozoites obtained from natural, chronic cases of caprine besnoitiosis produced clinical disease in goats but not in rabbits, mice, guinea pigs, hamsters, rats or cattle. Histological examination of tissue sections from the experimental animals showedBesnoitia cysts only in goats. This, together with field observations that cattle reared together with goats having besnoitiosis do not contract the disease, suggests that theBesnoitia species that infects goats in Kenya is host-specific and is notBesnoitia besnoiti. We suggest that the nameBesnoitia caprae be adopted for the caprine pathogen.     

间接酶联免疫吸附试验诊断牛贝诺孢子虫病的试验研究

由贝诺孢子虫病牛皮肤组织包囊内分离滋养体制备抗原。然后,通过正交试验筛选影响ELISA敏感性的几种主要因素,建立了诊断牛贝诺孢子虫病的问接酶联免疫吸附试验方法。对已知阳性和阴性血清各30头份进行检测,其符合率均为100％;对疫区244头份血清进行检测,其阳性检出率为34.8％,面临床通过巩膜险查的阳性检出率仅为4.5％;对巴贝贝西虫、双芽巴西虫、瑟氏泰勒虫、环形泰勒虫、伊氏锥虫、肉孢子虫等感染牛血清68头份进行检测,除22头份肉孢子虫惑染牛血清中有5头份判为阳性外,其余均为阴性。初步尝试可以全血于纸片代替血清进行检测,其检测结果与血清一致。对ELISA检测结果以肉眼判定,与OD值测定结果间的误差不超过3.?％。     

A survey on hydatidosis in livestock in Northern Iran based on data collected from slaughterhouses from 2004 to 2008

Data have been collected from slaughterhouses in three provinces across the Northern Iran (Gilan, Mazandaran and Golestan) from March 2004 to March 2008. These data were collected to evaluate the prevalence of hydatidosis in slaughtered cattle, sheep and goats. During the study, 3,347,797 animals were slaughtered. These included 621,686 cattle, 1,719,725 sheep and 1,006,386 goats. The prevalence of infection in cattle, sheep and goats was 12%, 14.6% and 10.1%, respectively. The association of condemnation rates with seasons was not proven statistically.