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1.
为了观察ICT人工消化法检验肉类中旋毛虫敏感性的影响因素与优化方案的检验效果.将120只小鼠随机分为3组(每组40只),分别经口感染300条、20条、5条旋毛虫感染性肌幼虫,42 d后剖杀.肌肉剪碎后经ICT-消化法消化,收集肌幼虫,观察不同沉淀容器、温度、过滤筛及消化时间对幼虫回收效果的影响.结果表明,幼虫回收率随筛孔径的增大而升高(p<0.01),随温度升高而降低(p<0.01).肉样消化2 h的幼虫回收率(96.18%)明显高于消化005 h(88%)(p<0.05).ICT-消化法与改良消化法用于5条旋毛虫感染小鼠肌肉检验时的阳性率均为100%,但后者收集的幼虫数(68.70条)明显多于前者(55.90条)(p<0.01);改良消化法对每克肌肉虫荷(1pg)为0.1 lpg和1 lpg的肉样检验时,敏感性与幼虫回收率均为100%.消化法检验肉类中旋毛虫的最佳方案为肉样43℃消化2 h、消化液冷却至4℃时40目(425 μm孔径)筛过滤及锥形量杯沉淀.  相似文献   

2.
为了观察小鼠感染不同种旋毛虫后肉汁抗体水平的变化及其与血清抗体水平的相关性,将200只昆明小鼠随机分成4组(每组50只),每只分别感染300条乡土旋毛虫(T2)、布氏旋毛虫(T3)、伪旋毛虫(T4)及纳氏旋毛虫(T7)幼虫,感染后2~6周每组每周随机剖杀10只小鼠,收集血清及肉汁,用旋毛虫肌幼虫ES抗原ELISA检测血清及肉汁中抗体水平;另取40只昆明小鼠随机分成4组(每组10只),每只分别感染500条T2、T3、T4、T7幼虫,感染后6周剖杀,制备肉样,用ELISA检测4℃及-20℃保存不同时间后的肉汁抗旋毛虫抗体动态。T2、T3、T4、T7感染小鼠后的肉汁抗体水平和变化趋势相似,均是在感染后第3周检测出肉汁特异性抗体,抗体阳性率分别为60%、30%、30%、30%,至感染后第4周肉汁抗体阳性率均升至100%。4组小鼠感染后2~6周的肉汁与血清抗体水平均具有相关性。T2、T3、T4、T7感染小鼠肌肉4℃保存7d与1d的肉汁抗体水平相比均无显著性差异;所有的感染小鼠肌肉-20℃保存2个月的肉汁抗体水平与保存1个月的相比均无显著性差异;虽然保存3个月的肉汁抗体水平与保存1个月的相比均已有显著性差异,但抗体阳性率仍均为100%并持续至实验结束时的4个月保存期。结果表明,肉汁中抗旋毛虫抗体的检测可用于新鲜肉、冷藏肉及冷冻肉中乡土旋毛虫、布氏旋毛虫、伪旋毛虫、纳氏旋毛虫捡疫的初筛。  相似文献   

3.
本研究旨在分析肌幼虫期旋毛虫感染小鼠肠道菌群的结构组成变化。选取12只雌性BALB/c小鼠,随机分为对照组(CK)和感染组(Ts35),感染组小鼠经口感染300条旋毛虫肌幼虫,对照组口服等量PBS。收集对照组和感染组小鼠在感染后第35 d的盲肠内容物,提取DNA,并对其进行16S rRNA测序及生物信息学分析。结果发现,对照组小鼠和感染组小鼠肠道菌群的Alpha多样性没有显著差异,而β多样性发生变化,表明两组的菌群结构不同。Metastats分析发现,与CK组相比,旋毛虫感染35 d后的小鼠肠道中产丁酸的罗斯拜瑞氏菌(Roseburia)和抗炎及组织修复相关的梭菌属(Clostridium)丰度显著升高(P<0.05),而拟杆菌(Bacteroides)(P<0.01)、乳酸杆菌(Lactobacillus)和Ralstonia(P<0.05)的丰度显著下降。同样地,LEfSe分析发现罗斯拜瑞氏菌的丰度明显升高(P<0.05)。综上表明,肌幼虫期旋毛虫感染小鼠肠道菌群的组成与对照组有明显的不同。  相似文献   

4.
旋毛虫感染小鼠对p46 000重组抗原的抗体应答   总被引:1,自引:0,他引:1  
分别以旋毛虫肌幼虫ES抗原和p46000重组蛋白作为抗原,对小鼠人工感染旋毛虫后的抗体应答进行了ELISA检测。结果表明,以肌幼虫200条/只经口感染小鼠后,肌幼虫ES抗原在感染后9d可检出抗体,并于感染后35~42d达到最高水平;应用重组抗原检测时,感染后10d可检出抗体,抗体水平略低于用ES抗原,但是其消长规律基本一致.而且与阴性血清相比差异明显;抗体在117d后仍维持于较高水平。  相似文献   

5.
为进一步探讨旋毛虫P53基因原核表达重组蛋白的免疫保护作用,本实验采用纯化的重组蛋白作为抗原免疫小鼠,免疫剂量为50μg/100μL,每隔10d免疫1次,共3次。末次免疫后10d,每只小鼠以200条旋毛虫肌幼虫攻虫。分别检查感染后7d小鼠肠道内成虫数量、体外培养雌虫所产新生蚴数量以及感染后35d肌肉荷虫量。结果显示:小鼠肠道内成虫、新生蚴和肌幼虫的减虫率分别为61.9%、53.45%和71.48%,表明旋毛虫表达P53基因的重组蛋白对小鼠产生了较好的免疫保护效果。  相似文献   

6.
通过在小鼠体内感染猪、犬旋毛虫肌幼虫50条,再用同源免疫和交叉免疫方法攻击感染旋毛虫肌幼虫200条,来检测其肌幼虫的减虫率。试验得到猪旋毛同源免疫的减虫率为87.47%,对犬旋毛虫的交叉免疫减虫率是86.47%,犬旋毛虫同源免疫的减虫率为83.70%,其对猪旋毛虫的交叉免疫是83.46%。试验表明:旋毛虫肌幼虫自然免疫具有较好的免疫保护作用,交叉免疫的保护率低于同源免疫。  相似文献   

7.
将单一性别、混合性别旋毛虫肌幼虫分别灌胃感染小鼠,新生旋毛虫幼虫尾静脉注射感染小鼠,并以未感染小鼠作对照,在感染后不同时间检测IL-2、IFN-γ、IL-4、IL-10、TGF-β的血清含量变化。结果显示,与对照组相比,在感染后前7 d,混合性别感染组IL-2含量降低,IL-10、TGF-β、IFN-γ含量升高,而IL-4含量无明显变化;感染后7~35 d,混合性别感染组小鼠的5种细胞因子含量均升高。与混合性别感染组相比,感染后前7 d,单一性别感染组小鼠IFN-γ和TGF-β含量较高,IL-2含量较低;在感染后7~35 d, 5种细胞因子含量较低。与对照组相比,在感染后前7 d,新生幼虫尾静脉注射感染组小鼠的IL-2、IFN-γ、IL-10、TGF-β含量升高,而IL-4含量无明显变化;感染后7~28 d,各细胞因子含量均显著性升高。与混合性别感染组相比,在模拟的对应时间点,新生幼虫感染组5种细胞因子变化无显著差异。结果表明,旋毛虫单一性别感染和混合性别感染引起的细胞因子变化在观察期间有较大差异,而新生幼虫感染和混合性别感染在对应时间点细胞因子变化相似。因此,我们认为单一性别感染和混...  相似文献   

8.
本试验通过差速超速离心法获得旋毛虫肌幼虫期胞外囊泡(Trichinella spiralis muscle larvae extracellular vesicles,Ts-ML-EVs),经透射电镜观察、纳米颗粒追踪分析、流式细胞术和Western blot鉴定。选择6~8周龄的健康雌性BALB/c小鼠,随机分为4组,每组8只。试验设计为PBS对照组(PBS组)、佐剂对照组(PBS+佐剂组)、旋毛虫肌幼虫期排泄分泌产物免疫组(Ts-ML-ES+佐剂组)以及旋毛虫肌幼虫期胞外囊泡免疫组(Ts-ML-EVs+佐剂组),分别取相应抗原与佐剂等体积混合采用多点皮下注射方式免疫小鼠。首免后第2,4周各加强免疫1次,剂量不变。三免后2周,每只小鼠灌胃300条旋毛虫肌幼虫,灌胃后6 d每组取2只剖杀,统计旋毛虫成虫减虫结果;攻虫35 d后所有小鼠全部处死,计算肌幼虫减虫率。每次免疫前、攻虫前以及处死前眼眶采血收集血清并保存,统一进行ELISA分析。结果显示:成功获得Ts-ML-EVs,随着免疫次数增加,小鼠血清中特异性IgG、IgG1、IgG2a、IgA和IgE抗体水平均显著上升,在三免后2周达到...  相似文献   

9.
《中国兽医学报》2019,(5):931-935
基于上转发光免疫层析(UPT-LF)技术,旨在建立UPT-LF快速检测猪抗旋毛虫IgG抗体的方法。采用间接法制备UPT-LF试纸条,通过共价偶联使二抗山羊抗猪IgG和上转发光纳米颗粒(UCP)结合,并加入样品垫和被检血清结合,以旋毛虫肌幼虫丝氨酸蛋白酶抑制因子基因(WM5)表达的蛋白WM5为猪抗旋毛虫IgG特异性结合抗原,WM5(2 g/L)与兔抗山羊IgG(0.5 g/L)喷点于分析膜上作为UPT-LF试纸条检测带(T)与质控带(C),该试纸条命名为Tsp-UPT-LF。Tsp-UPT-LF通过对95份阴性血清检测,确定Tsp-UPT-LF的Cutoff值为0.099。Tsp-UPT-LF和ELISA分别对273份旋毛虫感染猪血清样本进行检测,总符合率为92.31%,一致性系数Kappa(K)值为0.837,表明两种方法具有高度一致性。Tsp-UPT-LF检测旋毛虫、猪囊虫、华支睾吸虫感染的猪血清Tsp-UPT-LF呈现对旋毛虫良好的特异性。Tsp-UPT-LF检测感染50 000条旋毛虫肌幼虫28 d猪血清为阳性,T/C值0.109,122 d阳性最强,T/C值为0.207,至425 d时阳性值有所下降,抗旋毛虫IgG阳性率为71.4%。本研究建立了基于UPT-LF技术快速检测猪抗旋毛虫IgG抗体的方法,为猪旋毛虫即时感染检测、确保食品安全提供了一种可参考的检测方法。  相似文献   

10.
应用RAPD技术分析了玳瑁的遗传多样性。用20个随机引物对中国南海海域玳瑁7个个体的基因组DNA进行了PCR扩增,共扩增出1 351条DNA片段,平均每个个体扩增出193条条带。在检测到的193条条带中,多态性条带为69条,多态性条带百分比为35.8%,条带大小在200 bp~3 000 bp之间,7个个体间遗传距离为0.082 9~0.1813,平均遗传距离为0.132 7±0.029 9,表明中国南海海域玳瑁的遗传多样性水平较低,应加强该区域玳瑁种质资源的保护。采用类平均聚类法(NJTREE)构建了7个个体相互关系的分子聚类图,表明该7个玳瑁个体没有形成种群的分化。  相似文献   

11.
The red fox (Vulpes vulpes) is considered one of the main reservoir of Trichinella spp. in Europe. As limited information on Trichinella infection in wildlife of Hungary is available, 2116 red foxes, representing more than 3% of the estimated fox population of the country, were screened to detect Trichinella larvae by a digestion method. Trichinella larvae from the 35 positive foxes were identified by a multiplex PCR as Trichinella britovi (30 isolates, 85.7%), Trichinella spiralis (4 isolates, 11.4%), and Trichinella pseudospiralis (1 isolate, 2.9%). The true mean intensity of T. britovi, T. spiralis and T. pseudospiralis larvae in lower forelimb muscles was 23.6, 3.5 and 13.5larvae/g, respectively. T. spiralis was detected only in the southern and eastern regions. The non-encapsulated T. pseudospiralis was recorded for the first time in Hungary. Although the overall true prevalence of Trichinella infection in foxes was only 1.8% (95% confidence interval, CI=1.5-2.1%), the spatial analysis reveals different risk regions. In the north-eastern counties bordering Slovakia and Ukraine (21% of the Hungarian territory), the true prevalence of Trichinella infection is significantly higher than that observed in other regions (6.0%, CI=4.8-7.1%). In the southern counties bordering Croatia, Serbia and Romania (41% of the Hungarian territory), the true prevalence of Trichinella infection is moderate (1.4%, CI=1.0-1.8%). In the north-western and central counties (38% of Hungarian territory), the prevalence of Trichinella infection is significantly lower (0.2%, CI=0.1-0.4%) than that of the other regions. Based on the statistical analysis and the evaluation of epidemiological data, none of the counties can be considered free of Trichinella infection. In the past decade, Trichinella infection has been detected only in few backyard pigs, and only few wild boar-related autochthonous infections in humans were described. Nevertheless, these results highlight the need of the maintenance of a strict monitoring and control programmes on Trichinella infection in farmed and hunted animals of Hungary.  相似文献   

12.
In Sweden, the prevalence of Trichinella infection in domestic pigs has greatly decreased since the 1970s, with no reports in the past 4 years. However, infected wild animals continue to be found. The objective of the present study was to identify the species of Trichinella present in animals of Sweden, so as to contribute to the knowledge on the distribution area and hosts useful for the prevention and control of this zoonosis. In the period 1985-2003, Trichinella larvae were detected in the muscles of 81/1800 (4.5%) red foxes (Vulpes vulpes), 1/6 (16.7%) arctic fox (Alopex lagopus), 1/7 (14.3%) wolf (Canis lupus), 10/200 (5.0%) lynxes (Lynx lynx), 4/8000 (0.05%) wild boars (Sus scrofa), and 27/66 x 10(6) (0.000041%) domestic pigs. All four Trichinella species previously found in Europe were detected (Trichinella spiralis, T. nativa, T. britovi and T. pseudospiralis). The non-encapsulated species T. pseudospiralis was detected in three wild boars from Holo (Stockholm area) and in one lynx from Froso (Ostersund area), suggesting that this species is widespread in Sweden. These findings are consistent with those of a study from Finland, both for the unexpected presence of T. pseudospiralis infection and the presence of the same four Trichinella species, suggesting that this epidemiological situation is present in the entire Scandinavian region. The widespread diffusion of T. pseudospiralis in the Scandinavian region is also important in terms of it potential impact on public health, given that human infection can occur and the difficulties to detect it by the trichinelloscopic examination.  相似文献   

13.
Twelve Merino sheep were experimentally shown to be susceptible to infection with Trichinella spiralis or T. pseudospiralis by feeding on infected carcasses of mice or by oral intubation with recovered muscle larvae. The larvae recovered from the sheep showed variable tissue distribution. The diaphragm and tongue were most affected. The viability of the recovered larvae was confirmed by successful passage in mice. The reproductive capacity of T. spiralis in sheep was higher than that of T. pseudospiralis, and also higher than its reproductive capacity in C57BL/6J mice. The reproductive capacity of T. pseudospiralis in sheep at a lower dose was higher than that observed in mice. However at higher doses, it was significantly lower than that in mice. Therefore, it may be concluded that the sheep may be considered a suitable host for both species of Trichinella.  相似文献   

14.
Epidemiology of trichinellosis in Asia and the Pacific Rim   总被引:4,自引:0,他引:4  
The epidemiology of trichinellosis, species of Trichinella present and the food and eating habits of people affected in Asia and the Pacific Rim are reviewed with emphasis on Japan, China and Thailand. Trichinella seems to be prevalent throughout this region although outbreaks of trichinellosis have not been reported in some areas. Major outbreaks of the disease have been reported primarily in China and Thailand. This is the result of three factors: (1) China and Thailand are highly endemic areas for this parasite; (2) the two countries are well-organized and there is a public health system that enables precise reporting of disease outbreaks and (3) culinary habits provide many opportunities to eat undercooked meats. Trichinella found in Asia and the Pacific Rim includes both encapsulated species (Trichinella spiralis, Trichinella britovi, Trichinella nativa) and noncapsulated species (Trichinella pseudospiralis, Trichinella papuae). T. britovi, isolated in Japan, is a different genotype from the European strain. Therefore, the Japanese strain of T. britovi is designated Trichinella T9. Human trichinellosis caused by T. pseudospiralis has occurred in New Zealand and Thailand. Tasmania has had animal cases of T. pseudospiralis infection and animals with T. papuae infection have been found in Papua New Guinea. Economic losses due to Trichinella infection are not negligible in China, where there have been more than 500 outbreaks of human trichinellosis, affecting more than 20,000 people and causing more than 200 deaths. In Thailand, over the past 27 years, 120 outbreaks were reported involving nearly 6700 patients and 97 deaths. Japan has had fewer outbreaks and some sporadic cases have been attributed to imported infection.  相似文献   

15.
Enzyme immunoassays using the triple antibody enzyme linked immunosorbent assay (ELISA) with both Trichinella spiralis spiralis and T. spiralis nativa excretory-secretory (ES) antigens and a commercial Trichinella spiralis enzyme immunoassay test kit were carried out on sera from pigs that were infected with light, moderate and high doses of infective T. spiralis spiralis and T. spiralis nativa respectively. Seroconversion occurred in all pigs given infective Trichinella larvae although no trichinae were recovered from pigs given T. spiralis nativa larvae and examined between days 92 and 99 postinfection by pepsin digestion. Anti-Trichinella antibodies were detected in pigs infected with T. spiralis spiralis and T. spiralis nativa by ELISA using either the homologous or heterologous ES antigen. The commercial Trichinella spiralis enzyme immunoassay test kit also detected anti-Trichinella antibodies in both the T. spiralis spiralis and T. spiralis nativa infected pigs. The commercial test kit did not appear to be as sensitive as the triple antibody ELISA since it usually took two to three days longer for seroconversion to be detected by the former procedure. Finally seroconversion occurred more rapidly in swine infected with T. spiralis spiralis than with pigs receiving comparable doses of T. spiralis nativa.  相似文献   

16.
A wild boar (Sus scrofa) from the island Usedom in Mecklenburg-Western Pomerania (north-east Germany) was detected as Trichinella-positive during routine meat inspection. Encapsulated and non-encapsulated larvae were detected in the muscle tissue by trichinoscopy. In the diaphragm, 922 larvae per g were detected by artificial digestion. Muscle larvae displayed two different sizes of about 700 and 1100 microm. By a multiplex PCR analysis, larvae with a large size were identified as Trichinella spiralis, whereas those of a smaller size were identified as Trichinella pseudospiralis. This is the first finding of a mixed infection of T. spiralis and T. pseudospiralis in a naturally infected animal and it supports the tendency of more frequent detection of the non-encapsulated species T. pseudospiralis in Europe.  相似文献   

17.
A total of 72 specific pathogen-free (SPF) and Iberian pigs (three animals per group) were inoculated with 200, 1000 or 20,000 muscle larvae of T. spiralis, T. nativa, T. britovi and T. pseudospiralis. For each animal, the muscle larva burden was evaluated in nine muscle samples by digestion. The anti-Trichinella IgG kinetics in blood samples, taken twice prior and at days 5, 10, 15, 20, 25, 30, 40, 50 and 60 post-inoculation, and in muscle juice, obtained at necropsy, was evaluated by an ELISA using an excretory/secretory antigen. The mean larval recovery rate in SPF/Iberian pigs corresponded with the level of inoculum dose, and tongue, diaphragm and masseter were identified as predilection muscles. In SPF and Iberian pigs receiving 20,000 larvae of T. spiralis, an earlier seroconversion was detected from day 25 post-inoculation. At a 10-fold dilution, the muscle juice showed a good test agreement with blood serum.  相似文献   

18.
Our previous study showed that the IgA monoclonal antibody (mAb) HUSM-Tb1 forms immunoprecipitates on the cuticular surface of infective larvae of Trichinella britovi, and that intraperitoneal injection of this mAb to mice 5 hr before challenge infection confers a high level of protection against intestinal T. britovi. The same treatment produced a similar effect in BALB/c mice inoculated orally with Trichinella pseudospiralis larvae, indicating that the effects may be seen upon most members of the genus Trichinella. Worms recovered from the intestinal mucosa at 1 hr after challenge infection with T. pseudospiralis was few in mice passively immunized with the mAb, whereas a substantial number of worms were recovered from the mucosa of control groups. These results suggest that the IgA mAb impedes establishment of infective Trichinella worms in the intestinal mucosa. Trichinella worms inoculated orally into BALB/c mice vaccinated with ultraviolet-irradiated muscle larvae 3 weeks earlier were expelled between days 4 and 7 after challenge infection. Although the mAb HUSM-Tb1 originated from the mesenteric lymph node cells of mice vaccinated repeatedly with such irradiated larvae, IgA-mediated expulsion does not seem to play an important role in this vaccination model.  相似文献   

19.
Nematodes of the genus Trichinella are maintained in nature by sylvatic or domestic cycles. The sylvatic cycle is widespread on all continents, from frigid to torrid zones, and it is maintained by cannibalism and scavenging behavior of carnivores. Trichinella is primarily a parasite of carnivorous mammals, although one non-encapsulated species, Trichinella pseudospiralis, has also been detected in birds. The anaerobic metabolism of larvae in nurse cells allows their survival in extremely decayed meat. Encapsulated larvae in the decomposing carcass function similarly to the species-dispersing population of eggs or larvae of other nematodes, suggesting that the natural cycle of Trichinella includes a free-living stage when the parasite is no longer protected by the homeothermy of the host. Consequently, environmental temperature and humidity play an important role in the transmission of Trichinella among wildlife. Of the 10 recognized genotypes of Trichinella, only Trichinella spiralis is transmitted and maintained in a domestic cycle, although it can be present also in wildlife. All other genotypes (Trichinella nativa, Trichinella britovi, T. pseudospiralis, Trichinella murrelli, Trichinella nelsoni and Trichinella papuae, Trichinella T6, T8, and T9) are transmitted and maintained only in a sylvatic cycle. This generalization does not preclude sylvatic species of Trichinella from invading the domestic habitat, and T. spiralis may return to this habitat when humans fail in the management of wildlife and domestic animals. However, the presence of sylvatic genotypes of Trichinella in the domestic habitat represents a "dead-end" for the sylvatic cycle. Synanthropic animals (rats, foxes, mustelids, cats, dogs, etc.) contribute to the flow of sylvatic Trichinella genotypes from wildlife to domestic animals and of T. spiralis from domestic to sylvatic animals. Furthermore, human behavior not only influences the transmission patterns of Trichinella genotypes in the domestic habitat, but also it can contribute to the transmission and spread of this infection among wildlife, for example by improper hunting practices.  相似文献   

20.
Infection of Trichinella spp. is widespread among wildlife in Slovakia and the red fox (Vulpes vulpes) is the main reservoir of Trichinella britovi. Trichinella spiralis has been rarely documented in sylvatic and domestic animals of this country. During routine examination of domestic pigs at the slaughter, Trichinella larvae were detected by artificial digestion in a domestic pig of a large-scale breeding farm in Eastern Slovakia. The parasite has been identified by molecular (PCR) and biochemical (allozymes) analyses and by the morphology of the nurse cell as the non-encapsulated species Trichinella pseudospiralis infecting both mammals and birds. The epidemiological investigation carried out at the farm level revealed the presence of the same parasite species in other three pigs of 192 examined (2.1%), in 3 of 14 (21.4%) examined synanthropic rats (Rattus norvegicus) and in a domestic cat. The farm was characterized by inadequate sanitary conditions, insufficient nutrition, cannibalism and the presence of rat population. A different profile has been observed at the phosphoglucomutase locus in T. pseudospiralis isolates from Slovakia in comparison with the T. pseudospiralis reference isolate from the Palearctic region. This is the first documented focus of T. pseudospiralis from Central Europe. The detection in domestic pigs of a non-encapsulated parasite infecting both mammals and birds stresses the need to avoid the use of trichinelloscopy to detect this infection at the slaughterhouse.  相似文献   

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