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畜牧人

標題: 請各位老師幫我看看這篇翻譯 [打印本頁]

作者: 張家富 時間: 2008-5-9 14:06
標題: 請各位老師幫我看看這篇翻譯

請各位老師幫我看看這篇翻譯

這是我畢業前要交的一篇外文翻譯，請各位老師幫我看看并提出您寶貴的建議。非常感謝！

由于在這個貼子中不能上傳PDF格式的文件因此

英文原文上傳在http://www.cqnsfzw.com/plugin.php?identifier=download&module=download&acti=softview&softid=10057

由于下載是要花費大家的論壇幣的，我會在各位下載后把論壇幣返還給各位的。

并且對于提出修改建議的給予一定的論壇幣做為補賞您花費的時間。

以下是英文原文：

First report of Enterocytozoon bieneusi infection on a pig farm in the Czech epublic

Abstract
Enterocytozoon bieneusi infects humans and animals and can cause life-threatening diarrhea in immunocompromised people.
The routes of transmission and its zoonotic potential are not fully understood. Pigs have been frequently reported to have E.
bieneusi; therefore, we surveyed farm-raised pigs in the Czech Republic to determine its presence and genetic diversity. Spores were
detected by microscopy in the faeces of 65 out of 79 examined animals (82%). A species-specific polymerase chain reaction (PCR)
identified E. bieneusi in 94% of samples. Genotyping based on the ITS regions of the SSU rRNA gene identified that most pigs were
infected with the species-specific genotype F, while two animals had the zoonotic genotype D and two had genotype Peru 9. This is
the first report of E. bieneusi in swine in the Czech Republic, and demonstrated that most infections were with pig-specific
genotypes. Nonetheless, swine may still play a role in the transmission of E. bieneusi to humans.
# 2008 Elsevier B.V. All rights reserved.
Keywords: Enterocytozoon bieneusi; Pigs; Microsporidia; Genotypes; Zoonosis

1. Introduction
Of the 14 species of microsporidia currently known
to infect humans, Enterocytozoon bieneusi (Desportes
et al., 1985) is the most common and important cause of
human microsporidiosis associated with diarrhea and
systemic disease (Didier, 2005). The only published
case of microsporidiosis caused by E. bieneusi in AIDS
patient in the Czech Republic was recorded in 1994
(Ditrich et al., 1994).
Microsporidia spores could be released into the
environment via stool, urine, or respiratory secretions.
They are the infectious stage and can survive in different
environmental matrices such as water, soil, and food
products (Sinski, 2003). The spores of E. bieneusi have
been identified in surface waters in the USA and France
(Dowd et al., 1998; Cotte et al., 1999; Fournier et al.,
2000), and the possible sources of contamination were
humans or animals. Recently, E. bieneusi was also
detected in several wild, domestic and livestock animals
including swine, cattle, goats, birds, rabbits, dogs, cats
and macaques (Deplazes et al., 1996; Kondova et al.,
1998; Breitenmoser et al., 1999; del Aguila et al., 1999;
Mathis et al., 1999; Rinder et al., 2000; Dengjel et al.,
2001; Buckholt et al., 2002; Lores et al., 2002; Reetz
et al., 2002; Fayer et al., 2003; Santı´n et al., 2004, Ha
et al., 2005, Lobo et al., 2006). Sequence analysis of the
ITS region of the rRNA gene showed a close relationship
between E. bieneusi isolates from humans and pigs,
suggestive of an absence of transmission barriers between
these microsporidia isolates (Rinder et al., 2000).
The fact that E. bieneusi was identified in different
animals and water sources have raised public health
concerns about its potential as a zoonotic and waterborne
pathogen (Didier et al., 2004; Cama et al., 2007).
Although vertebrate hosts have been identified for this
microsporidium species infecting humans, the reservoirs
and the modes of transmission of E. bieneusi are
still unknown. Additionally, there is insufficient data to
understand the dynamics of microsporidia infections in
captive and farm animals in Central Europe, including
the Czech Republic.
We report the first survey on the occurrence and
prevalence of E. bieneusi infection on a pig farm in the
Czech Republic and detect its genotypes as a potential
source of human infection.
2. Materials and methods
2.1. Collection of stool samples
We conducted a coprological survey for microsporidia
in a randomly selected farm in the region of
Vysocˇina, Czech Republic during the autumn of 2006.
The selected farm had three different units: two
breeding complexes and a growing complex. Each
breeding complex was divided into two sections: one
with individual pens for furrowing sows and their litters
where piglets (pre-weaners) stay until weaned at 4
weeks of age. The second section was adjacent and
composed of large communal pens for weaned piglets
(starters), where they were kept until reaching 8 weeks
of age. Thereafter, piglets were transferred to the
growing complex (pre-growers), where they were kept
until about 12 weeks of age.
Faeces were collected from animals of different age
categories: pre-weaners, starters, pre-growers and sows.
We randomly sampled 10% of sows, 10% of pregrowers
and three piglets (pre-weaners or starters) per
litter of each sow in this study. The samples were
collected from the floor immediately after defecation
into individually labeled sterile tubes and stored at 4 8C
until processed in the laboratory.
2.2. Detection of microsporidium spores in faeces
Microsporidia were microscopically detected in
faeces using calcofluor white staining (Va´vra et al.,
1993). Briefly, thin smears were made from individual
stool samples and fixed in absolute methanol
and spores were visualized with 0.1% calcofluor
M2R stain (Sigma–Aldrich, St. Louis, MO, USA)
in phosphate-buffered saline (PBS). Evans blue
solution at 0.5% was used to facilitate differentiation.
The slides were examined using UV-light
with a filter wavelength of 490 nm and 1000 magnification.
2.3. Molecular characterization of microsporidia
species and genotypes
2.3.1. DNA extraction
Two to three hundred milligrams of faecal samples
from each sample were homogenized by bead disruption
using a Mini-BeadBeater (Biospec Products,
Bartlesville, OK, USA) for 120 s at a speed
5000 rpm. Total DNA was extracted using the
QIAamp1 DNA Stool Mini Kit (QIAGEN, Hilden,
Germany) following the manufacturer’s instructions
and was kept frozen at 20 8C until PCR amplification.
2.3.2. Molecular identification and
characterization of E. bieneusi
A nested PCR protocol that differentiates E.
bieneusi from other microsporidia commonly found
in humanswas used to amplify a 508 bp fragment of the
small subunit rRNAgene comprised of 122 bp of the 30-
end of the SSU rRNA gene, 243 bp of the ITS and
143 bp of the 50-region of the LSU rRNA gene
(Katzwinkel-Wladarsch et al., 1996). Briefly, primers
sets MSP-1 and MSP-2B, and MSP-3 and MSP-4B
were used for the primary and secondary PCR
amplifications, respectively. PCR amplification consisted
of 35 cycles, of 94 8C for 45 s, 52 8C for 45 s, and
72 8C for 60 s preceded by a denaturation step of 94 8C
for 3 min and followed by a final extension at 72 8C for
7 min. The amplicons were electrophoresed in 2%
agarose gels and visualized with 0.2 mg/ml ethidium
bromide.
All amplified products were sequenced in both
directions using BigDye Terminator chemistries
(Applied Biosystems, Foster City, CA, USA) using
the secondary primers MSP-3 and MSP-4B and
sequenced on the ABI PRISM 3031 (Applied Biosystems)
genetic analyzer. The resulting sequences were
assembled and aligned using the programs ChromasPro
Version 1.32 (Technelysium Pty. Ltd., Qld, Australia)
and Clustal X (ftp://ftp-igbmc.u-strasbg.fr/pub/ClustalX/)
and compared with reference sequences from
GenBank.
B. Sak et al. / Veterinary Parasitology 153 (2008) 220–224 221
2.4. Statistics
For statistical evaluation of results, Statistica1,
Release 5.1 Software (Statsoft, Tulsa, OK, USA, 1997)
was used. The Chi-square statistic test for evaluation of
significant differences were utilized.
3. Results
A total of 79 faecal samples were collected from 31
pre-weaners, 16 starters, 17 pre-growers and 15 sows.
At the time of collection, most animals appeared to be in
good health condition, and in two cases (pre-weaner and
pre-grower) diarrhea was inferred from the liquid
consistency of the stools. No association between the
occurrence of diarrhea and E. bieneusi infections in pigs
was found (x2 = 0.1386; df, 1; P > 0.05).
Microsporidia infection was detected in samples
from the three sections of the farm and in all age
categories (Table 1). Stool samples from 65 (82%) pigs
were microscopically positive (range 80–88%).
The PCR analysis of all stool samples detected E.
bieneusi in 74 (94%) of the animals, with very high
percentages among all age categories: all 31 pre-weaners,
15out of16starters, 16out of17pre-growersand12out of
15 sows (Table 1). Sequence analyses of the PCRamplified
product showed that themajority of E. bieneusi
detected in samples were 100% homological with
genotype F (GenBank accession number AF135833),
while four samples belonged to different genotypes: two
were 100% homological with genotype D (samples 50
and 52) and twowith genotype Peru 9 (samples 16 and 80)
(GenBank accessionnumbersAF101200 andAY371284,
respectively),whichwere previously reported in humans.
4. Discussion
In this work the specific diagnosis of E. bieneusi on a
closed pig farm in the Czech Republic was described.
We chose swine because they have been previously
reported to harbour human pathogenic genotypes of E.
bieneusi.
Surprisingly, we identified that 94% of the tested
pigs had E. bieneusi, and that this proportion was
similar among all age categories, and significantly
higher than any previously reported. In 1999, Breitenmoser
reported E. bieneusi in 35% of 109 pigs, and
with much higher occurrence among weaned piglets
(Breitenmoser et al., 1999). Similar to our results, most
of those isolates belonged to genotype F. In an 18-month
survey at a slaughter house in Massachusetts, 32% of
202 finished pigs had E. bieneusi: 18% of them had
microsporidia in their stool samples and the rest in
samples of bile (Buckholt et al., 2002). A small study of
six pigs suffering from severe diarrhea and stunting
identified that four of the six animals had microsporidiosis
(67%) (Rinder et al., 2000).
Although all studies do indicate that microsporidia,
and specifically E. bieneusi can infect swine, it is not yet
clear why there is a broad difference in prevalence
among the different studies. It can be suggested that the
observed differences were the result of different
farming practices from different parts of the world,
although all reported surveys including this one, used
samples from pigs that were raised under intense swine
production practices. These practices include having a
swine exclusive farm, age segregation and age-specific
feeding and enclosed facilities. Besides husbandry
practices, other factors that may affect the presence of
microsporidia in farmed animals can be the health
condition of the herd, age of the animals studied,
location and weather conditions at the farm.
Our results also suggest that it is very likely that
piglets may acquire the infection from their mothers at a
very early age, although the specific routes are unknown.
The faecal oral route has been proposed for acquiring E.
bieneusi infections, and transmission could have
occurred through direct contact with stools from the
sow, nursing from contaminated teats, or ingestion of
faecal material. Nevertheless, additional studies are
needed to ascertain the mechanics of this transmission.
Our study is the first to conduct a methodical survey
that discriminates pigs by age categories within
intensive farming conditions, while also collecting
samples from the sow and its litter. In this case, the close
proximity of the sow with her litter might have resulted
in an overestimation of E. bieneusi positive piglets,
which could have occurred as the result of contamination
with spores from the sow rather than actual
infection of the piglet. Nonetheless, the high percentage
of microsporidia positive animals persisted in piglets
that were weaned and physically separated from their
mothers, reinforcing our findings among pre-weaners.
Our findings also confirm that E. bieneusi genotype F
is the most frequently detected microsporidia in pigs
strengthening the concept of host specificity for this
genotype. Nonetheless, the detection of some genotypes
previously identified in humans suggests that pigs may
play a role, although minor, in the zoonotic transmission
of E. bieneusi.
Although pigs become infected at an early age and
excrete spores lifelong, microsporidia may remain
unrecognized because it is only rarely associated with
severe gastrointestinal symptoms and its detection by
microscopy requires specialized stains that are not part
of the routine coproparasitological diagnosis. Further
epidemiological studies are needed to fully ascertain its
distribution in different geographical settings, weather
conditions and husbandry practices, its economic
impact for the swine industry, and the zoonotic potential
of microsporidiosis in pigs in the Czech Republic.
Acknowledgements
We would like to thank the farm management and
employees to enable us to obtain samples.
This work was supported by the research project of
the Ministry of Education, Youth and Sports of the
Czech Republic (MSM 6007665806), by the grant of
the Grant Agency of the Czech Republic (project no.
523/07/P117) and research project of the Institute of
Parasitology, Biology Centre of the Academy of
Sciences of the Czech Republic (Z60220518).

Table 1
Frequency of E. bieneusi in different age categories of pigs on farms using light microscopy and genotyping
Age category Examination (positive/examined) Genotype F/D/Peru 9
Microscopy PCR
Breeding complex 1 Pre-weaners 19/21 21/21 21/0/0
Starters 10/13 13/13 13/0/0
Sows 7/11 9/11 9/0/0
Breeding complex 2 Pre-weaners 9/10 10/10 8/0/2
Starters 2/3 2/3 2/0/0
Sows 3/4 3/4 3/0/0
Growing complex Pre-growers 15/17 16/17 14/2/0

[ 本帖最后由張家富于 2008-5-9 14:32 編輯 ]

作者: 張家富 時間: 2008-5-9 14:25
標題: 譯文：很多詞沒有翻譯出來請各位老師幫忙
捷克共和國豬場的第一次Enterocytozoon bieceusi 感染報道
謫要：Enterocytozoon bieceusi感染人類和動物，并對免疫弱的人群造成腹瀉的生命危險，但對其傳染途徑及人獸共患的潛在性并不十分清楚。經常出現豬感染E. bieceusi的報道。于是我們在捷克共和國某個農場飼養豬群中檢測，以便確定病毒的存在及其遺傳多樣性。通過鏡檢術，在79只豬糞中檢測出65只豬的糞中含有病毒芽孢（82%）。在檢測中，一種特異性的聚合酶鏈反應（PCR）識別E. bieceusi的占94%。可辯出大部分豬感染了這種特異性的基因型F，其基因分型取決于小亞基rRNA基因ITS片段。雖然兩種動物含有人獸共患的基因型D，兩種含有基因型Peru 9。這是第一份在捷克共和國有關豬群感染E. bieceusi的報道。并且證實了大部分的感染帶有特異性的基因型。盡管如此，豬在E. bieceusi傳染給人的途徑仍然可能扮演重要角色。
關鍵詞：Enterocytozoon bieceusi；豬；微孢子目；基因型；人患共患病

1．說明
在當前已知的感染人類的微孢子目的14個種類中，Enterocytozoon bieceusi是造成與腹瀉、系統疾病有關的人類microsporidiosis的普遍和重要的原因。唯一出版過的由E. bieceusi造成的microsporidiosis事件，1994年發生在捷克共和國AIDS患者身上。微孢子目芽孢能通過糞便、尿液或者呼吸分泌物排泄到環境中，這些是傳染的階段，然后殘存于是同環境的基質中，如水、石油和食物成品中。E. bieceusi芽孢已在美國和法國地表水中檢測出，且可能成為人類或動物的污染源。近來E. bieceusi也在一些野生動物、家養的牲畜動物中檢測出，包括豬、牛、鳥、兔、狗、貓和macaque.rRNA基因ITS片段的數字分析顯示：在E. bieceusi從人類分離和從豬分離之前有密切聯系，在這些微孢子目隔離中，傳染障礙的不存在，讓人產生聯想。
其實，E. bieceusi已在不同種類動物和水資源中被檢測出，已經引起公共健康，關注穹成為人獸共患、水傳播的潛在的致病菌。雖然脊椎動物宿主已經被識別出這種感染人類的microsporidiosis，但貯主和E. bieceusi傳染模式仍然未知。此外，在捷克共和國等中歐地區，通過捕獲的和農場動物，只是利用不充分的數據去研究微孢子目的感染dynamics.
我們在這次不幸事件和在捷克共和國一個豬場E. bieceusi感染的流行事件中報道了第一次檢測，并檢測了它作為人類感染的潛在的根源的基因型。
2．材料和方法
2．1糞便取樣收集
我們在捷克共和國的2006年秋季期間，Vysocˇina，地區隨機選擇的一個農場，處理一個為微孢子目相對應的糞便學檢測。被選擇的農場分成三個單元：兩組繁殖組合和一組生長組合。每組繁殖組合被分成兩部分：一部分是單圈飼養面部有皺紋的經產母豬及其所產仔豬，仔豬（斷奶前仔豬）直到4周齡隔離，另一部分是毗鄰的和大部分共有圈組成，飼養斷奶仔豬直到8周齡。其后，生長豬被轉運到生長組合，直至大約12周齡。
來自不同年齡階段動物質糞便被收集：哺乳仔豬、斷奶仔豬、生長豬和繁殖母豬。這次的研究是：隨機采樣10%繁殖母豬，10%生長豬和每頭母豬每窩3只仔豬（哺乳仔豬或斷奶仔豬）。這些豬樣本在排糞后，立即從地板上收集后，分別裝入標記過的消毒的試管，然后貯存在實驗室4℃環境條件下直到下一步處理。
2．2糞便中microsporidium芽孢的檢測
微孢子目是在糞樣染色時使用calcofluor被微弱地檢測到的。簡單地說，薄層涂片是由各自的糞樣和純甲醇混合制成，芽孢在磷酸緩沖鹽溶液（PBS）中通過0.1% calcofluor M2R染色顯示，0.5%伊文氏藍溶液用以提高鑒別。載波片用帶有濾光片的490nm波長的紫外線在放在1000倍的條件下檢測。
2．3微孢子目種分子特征及其基因型
2．3．1DNA提取
所采取的糞樣各自用轉速為5000rpm/120s的小型球拌器通過破碎顆粒攪拌均勻，然后采樣200—300mg。接下來參照QIAamp DNA stool Mini Kit 產品說明書提取所有DNA，保存在-20℃條件下直至PCR擴增。
2．3．2分子識別和E. bieceusi特征
一種用于鑒別E. bieceusi和其它微孢子目的嵌套PCR方案普遍成立，在人類被用于擴增一個由末端為3’—的小亞基rRNA基因的122堿基對，ITS的243對堿基和LSU rRNA 基因的5’范圍的143對堿基構成的一個小亞基rRNA基因的508對堿基。總之，分別地將引物調定MSP-1和MSP-2B，MSP-3和MSP-4B用作首要和其次的PCR 擴增，PCR擴增在94℃條件下45s和72℃條件下60s循環35次，然后經過94℃條件下3分鐘的變性作用，緊接在72℃條件下7分鐘的最后擴散環節下完成。
所有擴增產物在兩個途徑下有序顯示，使用了BigDye 絡化學物利用，其次引物MSP-3和MSP-4B和在ABI PRISM 3031遺傳學analyzer。最后的有序結果和利用Chromas Pro Version 1.32和Clustal X排列相似，并和從基因庫參考的有序排列進行對比。
2．4統計
Statistica1,Release 5.1 Software 用來對結果的統計評估。The Chi-square統計用于有意義的區別評估。
3．結果
收集的79個糞樣，來自31頭哺乳仔豬、16頭斷乳仔豬和15頭繁殖母豬。在收集過程中，大部分動物處于健康狀況，在哺乳仔豬和斷奶仔豬時期，腹瀉的判定是依據糞便液體的一致性，且腹瀉的遭遇與E. bieceusi感染豬群沒有發現存在聯系。
微孢子目感染的檢測樣本，來自農場和所有年齡段分三部分，其中65頭豬（82%）的糞樣的微孢子目是呈陽性的（范圍80—88%）。
PCR分析，所有動物樣本中74只感染E. bieceusi（94%），在所有年齡分段中占很高比例：全部31只哺乳仔豬，15/16斷奶仔豬，16/17生長豬和12/15繁殖母豬。PCR擴增產物的有序分析顯示：在樣本中，大部分E. bieceusi 感染的是100%基因型F純合，4個樣本含有不同基因型：兩種100%基因型D純合和兩種含有基因型Peru 9 先前在人類感染報道過。
4．討論
我們的工作是在捷克共和國一個封閉的豬場研究E. bieceusi 的特異性特征。之所以選擇豬是因為先前報道了對人類有害的E. bieceusi 致病基因型。
令人意料不到的是，我們檢測的豬樣中94%含有E. bieceusi，且這個比例在所有年齡階段相似，更有意義的是比先前的任何報道都高。1999年，Breitenmoser 報道109只豬中含有E. bieceusi的占35%且遭遇較高的斷奶仔豬群中，和我們的結果相同之處是，大部分分離病毒屬于基因型F。在Masschusettes的一個屠宰室長達18個月的調查中202頭屠宰豬中32%含有E. bieceusi，18%在糞便采樣中檢測到微孢子目，余下的在膽汁采樣中檢測到。一項小研究，在6頭遭受嚴重腹瀉和功能喪失的豬樣中，檢測出有4頭帶有microsporidiosis。
盡管所有研究顯示微孢子目和特異的E. bieceusi 能感染豬，但是為什么在不同的研究中有關流行方面有多大的區別并不清楚。盡管所有報道的檢測包括一條，使用的樣品來自飼養豬在緊張狀態下的排泄物的常規。但它意味著觀測不同結果是由世界各地不同農場的慣例所造成。這些慣例包括：專有豬的農場，年齡分段和階段飼養技術和被固定的設備，此外還有管理。其它可能影響農場動物帶有微孢子目的因素：畜群的健康狀況，研究動物的年齡及農場的地理位置和氣候條件。
我們的結果也意味著，盡管不清楚感染途徑，但仔豬很可能在很小的時候從母豬那獲得感染。糞便的口服途徑已經被看成獲得E. bieceusi感染，傳染發生于直接接觸母豬糞便，哺乳時來自污染乳頭，或糞便物質的攝入。于是，額外的研究是需要查明傳染的方法。
我們的研究是第一次進行一個有條理的辨別檢測，即在一個集約豬場條件下通過年齡分段，應該收集的樣品來自繁殖母豬及其仔豬。在這樣的實施下，繁殖母豬及其仔豬的嚴格相鄰，致使出現一批E. bieceusi 陽性過高的仔豬，其可能產生的原因為帶孢子的污染物，是來自繁殖母豬的豬群中，這種微孢子目高比例陽性動物持續，也增強了我們在哺乳仔豬群的調查結果。
我們的發現階也證實了E. bieceusi基因型F是在豬群中大多經常檢測到的微孢子目。增強了這種基因型宿主特異性的理念。盡管如此，先前在人類被辯認的一些基因型的檢測暗示豬在E. bieceusi人獸傳染中，扮演了較小但重要的角色。盡管豬在很小的時候感染，一生排泄芽孢，但微孢子目可能仍然不被認知，因為它只有極少才聯系到嚴重胃腸的癥狀，它的感染通過特定染色后鏡檢斷定，染色劑不是日常coproparasitological 的組成。進一步的流行病學研究需要完全確定在不同地理位置，氣候條件和管理常規中的分布，它對養豬行業的經濟影響和在捷克共和國豬群microsporidiosis的人獸共患的潛在性。

作者: 張家富 時間: 2008-5-9 14:30
把不會的詞先集中一下，主要要幾下幾個：Enterocytozoon bieceusi
E. bieceusi
microsporidiosis
macaque.rRNA
dynamics
Vysocˇina
microsporidium
calcofluor
QIAamp DNA stool Mini Kit
BigDye
ABI PRISM 3031
analyzer
Chromas Pro Version 1.32
Clustal X
Statistica1,Release 5.1 Software
The Chi-square
Breitenmoser
Masschusettes
coproparasitological

作者: bonderic 時間: 2008-5-9 14:47
Enterocytozoon bieceusi  比氏腸胞蟲病
E. bieceusi 比氏腸胞蟲病
microsporidiosis 微孢子蟲病
macaque.rRNA  獼猴 rRNA
dynamics 動力學
Vysocˇina　摩拉維亞山區　（是一個地名）
microsporidium　微孢子蟲
calcofluor　卡爾科弗盧爾熒光染色劑
QIAamp DNA stool Mini Kit
BigDye
ABI PRISM 3031
analyzer
Chromas Pro Version 1.32
Clustal X
  Statistica1,Release 5.1 Software
The Chi-square
Breitenmoser
Masschusettes
coproparasitological
查著，我覺得這些你不會的生詞完全可以通過百度或者是google來查出來，我也沒必要一個個幫你查。你在查這些生詞的同時一些句子的翻譯你自然就會了。多用借助一些網絡工具。

作者: leiforever 時間: 2008-7-2 21:55
看來樓主用google翻譯的？

作者: yy0746 時間: 2008-7-8 15:50
給你個建議：用CNKI的翻譯助手翻譯單詞會相對專業一點。

歡迎光臨畜牧人 (http://www.cqnsfzw.com/)