Trichinella zimbabwensis n.sp. (Nematoda), a new non-encapsulated species from crocodiles (Crocodylus niloticus) in Zimbabwe also infecting mammals

Since 1995, Trichinella larvae have been detected in 39.5% of farmed crocodiles (Crocodylus niloticus) in Zimbabwe. Morphological, biological, biochemical and molecular studies carried out on one isolate from a farmed crocodile in 2001 support the conclusion that this parasite belongs to a new species, which has been named Trichinella zimbabwensis n.sp. This species, whose larvae are non-encapsulated in host muscles, infects both reptiles and mammals. The morphology of adults and larvae is similar to that of Trichinella papuae. Adults of T. zimbabwensis cross in both directions with adults of T. papuae (i.e. male of T. zimbabwensis per female of T. papuae and male of T. papuae per female of T. zimbabwensis), producing F1 offspring which produce very few and less viable F2 larvae. Muscle larvae of T. zimbabwensis, like those of T. papuae, do not infect birds. Three allozymes (of a total of 10) are diagnostic between T. zimbabwensis and T. papuae, and five are diagnostic between T. zimbabwensis and Trichinella pseudospiralis, the third non-encapsulated species. The percentage of the pairwise alignment identity between T. zimbabwensis and the other Trichinella species for the cytochrome oxidase subunit I gene, the large subunit ribosomal-DNA (mt-lsrDNA) gene and the expansion segment five, shows that T. zimbabwensis is more similar to the two non-encapsulated species T. papuae (91% for cytochrome oxidase I; 96% for mt-lsrDNA; and 88% for expansion segment five) and T. pseudospiralis (88% for cytochrome oxidase I; 90% for mt-lsrDNA; and 66–73% for expansion segment five) than to any of the encapsulated species (85–86% for cytochrome oxidase I; 88–89% for mt-lsrDNA; and 71–79% for expansion segment five). This is the first non-encapsulated species discovered in Africa. The finding of a new Trichinella species that infects both reptiles and mammals suggests that the origin of Trichinella parasites dates back further than previously believed and can contribute to understanding the phylogeny and the epidemiology of the genus Trichinella.     

Trichinella nativa and Trichinella T9 in the Hokkaido island, Japan

Trichinella sp. muscle larvae were isolated from the thigh muscle of two red foxes (Vulpes vulpes) captured in Sapporo and Otofuke, Hokkaido, Japan, in 2003. Multiplex PCR designed for genotyping the genus Trichinella revealed that the Sapporo isolate showed a specific pattern to T. britovi complex (T. britovi, Trichinella T8 and Trichinella T9) and the Otofuke isolate showed that to T. nativa. Nucleotide sequences of a part of the mitochondrial cytochrome oxidase subunit I (COI) gene and internal transcribed spacer 2 (ITS2) of the Sapporo isolate showed the highest similarity to those of Trichinella T9, a species detected in the mainland of Japan. This study shows that both T. nativa and Trichinella T9 are circulating in wildlife of the Hokkaido island.     

A comparison of the antigenic characteristics of three geographical isolates of Trichinella

The antigenic characteristics of isolates of Trichinella from Alaska, Canada and Kenya (arctic, north temperate and tropical, respectively) were compared using two dimensional crossed immunoelectrophoresis. It was found that there were some differences in the antigenic components of the three isolates but little difference in the number of antigens. The usefulness of serology in taxonomy is discussed in relation to these three isolates.     

Trichinella spiralis and Trichinella pseudospiralis induce collagen synthesis by host fibroblasts in vitro and in vivo

Immunoperoxidase staining of muscle infected with Trichinella spiralis for murine collagen types I and IV provided both qualitative and quantitative evidence of extensive synthesis of both types of collagen by fibroblasts in infected muscle compared to that seen uninfected muscle. Moreover, fibroblasts in muscle infected with T. pseudospiralis, a nonencapsulating species, showed significantly less staining for both types of collagen compared to muscle from mice infected with T. spiralis. Analysis of collagen composition of isolated nurse cells using an ELISA specific for either type I or type IV murine collagen suggested that of these 2 types of collagen, only type IV basement membrane collagen is found in Trichinella capsular collagen. Excretory/secretory products of T. spiralis and T. pseudospiralis induced extensive synthesis of exclusively type IV collagen by 3T3 murine fibroblasts in vitro.     

Trichinellosis of animals in Chukotka, USSR: A general review

During an examination, for trichinellosis, of 1489 mammals of 12 different species, in the territory of Chukotka National district (regions of Anadyr', Bering, Iul'tin, Chaun and Chukotka) and in the region of North-Even of Magadan province, Trichinella larvae were detected in 236 (16·4%) of these animals from 6 of the species: in 109 (44·8%) of the blue polar foxes kept in cages, in 12 (15·8%) of the silver-black foxes kept in cages, in 84 (18·2%) of the white polar foxes, in 17 (17·5%) of the common foxes, in 13 (12·1%) of the dogs and in the one European bear examined. Trichinella larvae were not found in 190 marine mammals examined: 178 ringed seals, 7 whales, 4 walruses, and 1 bearded seal, nor in the 8 Siberian squirrels and 159 domestic pigs examined. The high incidence of Trichinella infection amongst blue polar foxes and silver-black foxes on fur-animal farms in Chukotka, where the food ration of these animals is predominately the flesh of marine mammals (walruses and other pinnipeds), suggests that marine mammals may constitute one of the sources of infection to caged animals. In Chukotka, as in other regions of the Extreme North and North-East, the circulation of trichinae in nature may occur between two ecologically different groups of animals: terrestial and marine mammals. A wide distribution of trichinellosis among animals in nature, or kept in cages and among dogs in Chukotka National district and in the North-Even region of Magadan province provides a constant threat of infection to man. Thus the flesh and sub-products of potential hosts of trichinae (including dogs, fur animals, European and polar bears, marine mammals, etc.) should undergo thorough trichinelloscopy examination before being consumed by man. The flesh of sea mammals should be examined for trichinellosis before being used as food for animals; the presence of trichinae in it requires its sterilization.     

Detection of Trichinella spiralis in a horse during routine examination in Italy

Routine examination for Trichinella infection by artificial digestion of 5-g samples of muscle tissue revealed the presence of muscle larvae in one out of 28 borses imported from Romania to an abattoir in Italy. The parasite, identified as Trichinella spiralis by the polymerase chain reaction, showed a reproductive capacity index of 68 in Swiss mice. Light microscope examination of 200 nurse cell-larva complexes showed that 22% of them were calcified and that the capsules of the non-calcified nurse cells were 17.5–27.5 μm (s=22.67 μm) thick and had very few cellular infiltrates. The serum samples from the parasitologically positive horse and from three other horses of the same stock, from which Trichinella larvae were not recovered by digestion, showed a low level of positivity as determined by ELISA and Western blot analyses using a crude antigen, whereas negative results were observed in both tests when an excretory-secretory antigen was used. The results, together with data from the literature, suggest that the horse had acquired the infection 8–10 months previously and confirm earlier observations obtained from experimental infections, which showed that muscle worm burden and specific circulating antibodies were lost several months after infection.     

Hypoxic pulmonary vasoconstriction in reptiles: a comparative study of four species with different lung structures and pulmonary blood pressures

Low O2 levels in the lungs of birds and mammals cause constriction of the pulmonary vasculature that elevates resistance to pulmonary blood flow and increases pulmonary blood pressure. This hypoxic pulmonary vasoconstriction (HPV) diverts pulmonary blood flow from poorly ventilated and hypoxic areas of the lung to more well-ventilated parts and is considered important for the local matching of ventilation to blood perfusion. In the present study, the effects of acute hypoxia on pulmonary and systemic blood flows and pressures were measured in four species of anesthetized reptiles with diverse lung structures and heart morphologies: varanid lizards (Varanus exanthematicus), caimans (Caiman latirostris), rattlesnakes (Crotalus durissus), and tegu lizards (Tupinambis merianae). As previously shown in turtles, hypoxia causes a reversible constriction of the pulmonary vasculature in varanids and caimans, decreasing pulmonary vascular conductance by 37 and 31%, respectively. These three species possess complex multicameral lungs, and it is likely that HPV would aid to secure ventilation-perfusion homogeneity. There was no HPV in rattlesnakes, which have structurally simple lungs where local ventilation-perfusion inhomogeneities are less likely to occur. However, tegu lizards, which also have simple unicameral lungs, did exhibit HPV, decreasing pulmonary vascular conductance by 32%, albeit at a lower threshold than varanids and caimans (6.2 kPa oxygen in inspired air vs. 8.2 and 13.9 kPa, respectively). Although these observations suggest that HPV is more pronounced in species with complex lungs and functionally divided hearts, it is also clear that other components are involved.     

深圳大鹏湾六种珊瑚共附生真菌多样性研究

本研究使用6种不同培养基对深圳大鹏湾6种珊瑚上的共附生真菌进行平板涂布法分离培养,结合ITS-rDNA基因序列特征和形态学特征进行鉴定来研究深圳大鹏湾海域珊瑚共附生真菌的多样性。共分离培养出457株真菌菌株,分属于7个属,分别为红酵母属Rhodotorula（180株）、曲霉属Aspergillus（170株）、木霉属Trichoderma（50株）、青霉属Penicillium（34株）、枝孢属Cladosporium（21株）、透孢黑团壳属Massarina（1株）和弯颈霉属Tolypocladium（1株）,其中优势属为红酵母属Rhodotorula和曲霉属Aspergillus,占菌株分离总数的76.59%。单独鹿角珊瑚上分离到的共附生真菌数量和种类最多,为160株9种;在该种珊瑚上还分离出弯颈霉属Tolypocladium sp.,该属真菌此前从未在珊瑚共附生真菌的研究中分离出来。本研究还发现SDA培养基分离出的共附生真菌数量最多,CDA培养基的真菌数量最少,表明不同培养基分离的共附生真菌多样性存在差异。     

一株南海指海绵共附生真菌多样性及其抗菌活性

本研究通过采用6种不同的培养基对南海指海绵上的共附生真菌进行平板涂布法分离培养,结合ITS-rDNA基因序列特征和形态学特征进行鉴定来研究海绵共附生真菌多样性。共分离培养出55株(21种)真菌菌株,优势菌属为木霉菌属Trichoderma sp.(31株),其中的哈茨木霉Trichoderma harzianum 22株作为优势菌种,占真菌菌株总数的40%。其余真菌则属于枝孢菌属Cladosporium、迈耶氏酵母属Meyerozyma、茎点霉属Phoma、红酵母属Rhodotorula、青霉菌属Penicillium、曲霉属Aspergillus等10个属,还有两株未鉴定到种的子囊菌。选取金黄色葡萄球菌Staphylococcus aureus等7种病原指示菌,采用琼脂扩散法对21种真菌的代表性菌株进行抗菌活性筛选,其中有3种真菌至少对一种指示菌表现出抑菌效果,占菌株种类总数的11%。     

Specificity of the interrelations of Leishmania and host cells in vitro

Experiments on cross infection of peritoneal macrophages of mice with Leishmania of reptiles L. gymnodactyli and free cells of abdominal cavity of caucasian Agama (some part of which is composed by fibroblasts) with Leishmania of mammals L. major and L. donovani have shown the possibility of reproduction of the above species both in reptiles and mammals. The persistence of L. gymnodactyli and L. major in macrophages of mice was traced up to 10 days, the abundance of L. gymnodactyli during the whole period of observations being lower than that of L. major. The abundance of the above Leishmania in these cells happened to be higher than in the cells of reptiles. In the cells of reptiles the infection with these three species of Leishmania was eliminated by 5-6 days. More activity internalization of Leishmania of reptiles into cells of reptiles as compared to Leishmania of mammals was revealed that, apparently, reflects a definite degree of their adaptation to existence in reptiles in vivo.     

健康与患病刺梨植株可培养叶际真菌菌群差异比较

本研究比较了健康与患叶斑病刺梨植株叶际真菌群落特征差异,以期探索病原菌的潜在来源,为人工构建具拮抗功能群落和刺梨叶斑病的生物防控提供参考。通过可培养方法对不同健康状况的刺梨叶际真菌进行分离培养,结合形态学和分子系统学对菌株进行综合鉴定;利用FUNGuild平台对真菌进行功能注释;结合根际、根部真菌作拆分网络分析探索病原菌的潜在来源。本研究结果表明：1）刺梨叶际真菌具有丰富的多样性。从刺梨叶际8个样品中共分离到真菌266株,其隶属于3门、6纲、13目、30科、46属中的61个种。其中,健康植株叶际内生真菌（LHE）包括8属10种27株,附生真菌（LHS）包括33属37种77株。患病叶际内生真菌（LDE）分离到7属10种38株;附生真菌（LDS）分离到31属35种124株。2）不同样品的真菌优势属和特有类群有差异。不同健康状况下叶际附生真菌的优势属均为拟盘多毛孢属Pestalotiopsis,但二者的相对多度存在差异,LHS为11.49%,LDS为32.26%;内生真菌优势属二者均为链格孢属Alternaria,但相对多度各异,LHE为33.33%,LDE为63.16%。其中,LHE特有类群为盘长孢状刺盘孢 Colletotrichum gloeosporioides和果生刺盘孢Colletotrichum fructicola等8种;LDE特有类群为茄链格孢 Alternaria solani和Didymella sinensis 等8种;LHS特有类群是草酸青霉Penicillium oxalicum和Peniophora crassitunicata等21种;LDS特有类群是尖孢镰刀菌Fusarium oxysporum、赭绿青霉Penicillium ochrochloron和易脆毛霉Mucor fragilis等19种。3）不同样品叶际真菌功能不同。经FUNGuild解析表明,LHS、LHE和LDE的叶际真菌功能群主要以腐生型为主,LDS则主要以植物病原菌群为主。本研究结果初步揭示刺梨植株健康与患叶斑病叶际间真菌多样性、群落组成及营养功能群存在差异,植株健康状况与其真菌群落特征密切相关;叶斑病病原菌主要源于刺梨叶际的附生微生物群。     

There are at least three genetically distinct small piroplasms from dogs

The 18S nuclear subunit ribosomal RNA (18S rRNA) gene of small piroplasms isolated from dogs from Okinawa (Japan), Oklahoma, North Carolina, Indiana, Missouri, and Alabama, was isolated and sequenced. Phylogenetic analysis of these sequences and comparisons with sequences from other Babesia, Cytauxzoon, and Theileria species revealed that all canine small babesial isolates, with the exception of isolates from California and Spain, were placed in a group containing the Babesia spp. sensu stricto. Within the Babesia spp. sensu stricto, there was support for separating the small canine piroplasms from the large canine piroplasm, Babesia canis. The isolate from California was in a distinct phylogenetic clade, closely related to babesial isolates from wildlife and humans from the Western US. The canine isolate from Spain was closely related to Babesia microti. These results suggest that there are multiple small piroplasm species in dogs. The isolates from the Midwestern and Eastern US and the one from Japan probably represent a single species with wide geographic distribution.     

通过红色名录评估探讨中国爬行动物受威胁现状及原因

为了评估中国爬行动物红色名录, 我们按照爬行动物生物学特性, 适当修改了IUCN濒危物种红色名录标准Version 3.1。在此基础上, 评估了中国爬行动物生存现状, 参与编制了《中国生物多样性红色名录——脊椎动物卷》。此次评估的中国爬行动物有3目32科133属461种, 结果为: 区域灭绝(RE) 2种、极危(CR) 34种、濒危(EN) 37种、易危(VU) 66种、近危(NT) 78种、无危(LC) 175种以及数据缺乏(DD) 69种。中国受威胁(包括极危、濒危、易危)的爬行动物共计137种, 约占总数的29.72%, 包括龟鳖目31种、有鳞目蛇亚目67种、蜥蜴亚目38种和鳄形目1种, 高于2014年《IUCN濒危物种红色名录》评估的世界爬行动物受威胁比例(13.61%)。在所有受威胁物种中, 受威胁比例最高的类群是鳄形目(100%)和龟鳖目(91.18%), 其次是有鳞目蛇亚目(28.39%), 第三是有鳞目蜥蜴亚目(20.21%)。中国爬行动物特有种受威胁物种有39种, 占特有种总数(143种)的27.27%, 占受威胁物种总数(137)的28.47%。长江以南的华南和西南地区受威胁的物种最多。爬行动物受人类干扰严重, 主要表现为: 栖息地质量退化及生境破碎化、过度利用及污染和气候变化等。尽管自1989年《中华人民共和国野生动物保护法》实施以来, 一些中国濒危爬行动物的生存状况得到改善。但鉴于中国爬行动物区系的独特性和多样性、地形地貌的复杂性及社会经济发展的不均衡性, 为了维持区域生态安全和资源可持续利用, 拯救中国濒危爬行动物, 尤其是中国特有爬行动物中的极危物种, 是中国动物保护工作最迫切的任务之一。     

鄂西地区蜜环菌生物种的遗传多样性研究

蜜环菌属Armillaria真菌是世界范围内广泛分布的担子菌,既是引起森林病害的重要病原菌,也是天麻和猪苓生长的必须共生真菌,具有重要的经济价值。本文对我国鄂西天麻主产区即神农架林区、大老岭自然保护区、宜昌市所属的夷陵区和五峰县、恩施自治州等地进行了系统的调查采样,共获得45份蜜环菌子实体样本及其二倍体菌株,其中通过孢子印分离获得约320个单孢菌株。在此基础上,采用IGS、ITS和elongation factor-1 alpha（EF1-α）分子标记对45份二倍体菌株进行测序和系统发育分析,对有单孢株的样本采用遗传互交不育性实验进行生物种鉴定。结果表明,鄂西地区的蜜环菌分为4个clade,共有6个不同的蜜环菌生物种,分别为CBS J,CBS L,CBS N,CBS O,CBS F和Armillaria mellea,具有丰富的遗传多样性。其中,24个样本鉴定为CBS L,是鄂西地区的优势种;9个样本鉴定为CBS J;9个样本鉴定为CBS N;另有3个样本分别鉴定为CBS O,CBS F和Armillaria mellea。其中CBS L、CBS N、CBS O首次在湖北地区报道,扩大了这3个生物种的分布范围。本研究为了解鄂西地区蜜环菌的资源情况提供基础数据,对天麻生产蜜环菌菌种选育具有重要的参考价值。     

Comparison of the Host Ranges and Antigenicity of Cryptosporidium parvum and Cryptosporidium wrairi from Guinea Pigs

ABSTRACT. Oocysts of a Cryptosporidium isolate from guinea pigs were not infectious for adult mice, but were infectious for two of three newborn calves and for suckling mice. However, oocysts isolated from calves or mice infected with guinea pig Cryptosporidium were not infectious for guinea pigs. Four isolates of C. parvum from calves were incapable of infecting weanling guinea pigs. Microscopic examination of tissue from the colon and cecum of suckling guinea pigs inoculated with C. parvum revealed sparse infection of some pups. These host range studies and previously described differences in ¹²⁵I-labeled oocyst surface protein profiles between Cryptosporidium sp. from guinea pigs and C. parvum suggest they are distinct species. We propose the name Cryptosporidium wrairi be retained. Studies with monoclonal antibodies indicate that C. wrairi and C. parvum are antigenically related.     

从自然保护区到国家公园体制试点: 三江源国家公园环境覆盖的变化及其对两栖爬行类保护的启示

三江源地区具有丰富的高原生物多样性, 是我国重要的生态安全屏障。作为我国第一个体制试点的国家公园, 三江源国家公园将被建成青藏高原大自然保护展示和生态文化传承区。为更好地服务于三江源国家公园建设, 本研究从海拔、年均温和年降水等方面, 在环境空间上系统比较了国家公园与三江源和可可西里自然保护区的差异; 以4种两栖爬行动物即高原林蛙(Rana kukunoris)、倭蛙(Nanorana pleskei)、西藏齿突蟾(Scutiger boulengeri)和青海沙蜥(Phrynocephalus vlangalii)分布点环境条件在国家公园内外的异同为例, 探讨分布受环境制约大、扩散能力弱的物种保护在国家公园建设中可能面临的机遇和挑战。结果表明, 国家公园与已建自然保护区在地理空间上重叠较大, 但它们在自然环境上的差别很明显; 自然保护区内两栖爬行类所偏好的环境条件较多未包含在国家公园内。鉴于三江源的区位特殊性及对气候变化的敏感性, 区域内两栖爬行动物等对环境依赖性强、扩散能力弱物种的有效保护不仅有助于保持物种遗传多样性和区域生态系统的完整性, 也有利于更好地实现国家公园目标定位并服务于生态文明建设。为此, 在国家公园内, 应开展两栖爬行动物种群动态和群落结构的长期监测, 加强基础生物学研究, 掌握环境变化对两栖爬行动物分布、遗传、行为、形态、种群动态及群落可能产生的影响, 实现区域内两栖爬行动物及其类似物种在环境变化下的永续生存。     

ITS/CHS1靶位序列分析结合SRAP分子标记技术鉴定Nannizzia incurvata临床分离株

以转录间隔区（internal transcribed spacer,ITS）和几丁质合成酶1（chitinase synthase 1,CHS1）为鉴定靶位,结合种系进化分析将2007年至2016年分离自头癣或面癣患儿的12株石膏样小孢子菌复合体成员（现被归入Nannizzia菌属）鉴定到种水平,其中5株为Nannizzia gypsea（Microsporum gypseum）,另7株为Nannizzia incurvata（Microsporum incurvatum）。在此基础上,通过序列相关扩增多态性（SRAP）分子标记进一步证实二者在DNA多位点存在扩增多态性。本研究证实在我国湖北地区存在Nannizzia incurvata,为我国开展Nannizzia菌属的分子流行病学研究提供了实验室依据和支持。     

Larval feeding performance of two Neotropical Ornithodoros ticks (Acari: Argasidae) on reptiles

This paper determines the feeding performance of the larvae of two Neotropical soft tick species namely Ornithodoros rostratus Aragão, 1911 and O. puertoricensis Fox, 1947 on reptiles (Gekkonidae) using rabbits, mice and guinea pigs to provide comparisons with feeding features on mammals. O. puertoricensis produced a larval feeding rate of 63% on reptiles, while that of O. rostratus was only 20%. But the final success (attaching + feeding) was similar, 12.4% for O. puertoricensis and 10.4% for O. rostratus. The feeding time was also very different for both species. In O. puertoricensis, detachment begins at 16th day and lasts until day 27. In O. rostratus detachment begins at 1.5 h and lasts until day 10. These values of feeding on reptiles are different from those obtained on mammals (average 5.6 days for O. puertoricensis and 2.9 for O. rostratus).     

Elucidation of Schistosoma japonicum population dynamics in pigs using PCR-based identification of individuals representing distinct cohorts

The study reported here investigated the interactions of successive infections and acquired resistance of pigs to challenge infections of Schistosoma japonicum. Two morphologically indistinguishable geographical isolates from China (from Anhui and Zhejiang provinces) were used for the infections. The worms of the two isolates were distinguishable by PCR-linked restriction fragment length polymorphism analysis of the nicotinamide adenine dinucleotide phosphate dehydrogenase I gene of the mitochondrial genome. Thirty-two pigs divided into seven groups were used in the experiment. Two groups received a single infection by either the Anhui or the Zhejiang isolate. In Challenge Groups 1, 4, 6, 8 and 12, a primary infection of the Zhejiang isolate was followed by a challenge infection with the Anhui isolate at week 1, 4, 6, 8 or 12 after the primary infection. In this way it was possible to determine whether worms recovered by perfusion originated from the primary or the challenge infection. Only the challenge infection at week 1 resulted in a higher worm burden when compared with a single primary infection with the Zhejiang isolate. The results showed that challenge worms were able to establish, and that the proportion of worms originating from challenge infection increased at the later challenge infections, however without an increase in the total number of worms. In addition, mixed pairs of the two isolates were found in all challenge-infected groups. The results indicate that pigs are able to mount a partial resistance against re-infection with S. japonicum by 4 weeks after a primary infection, but that worms of the challenge infections eventually replace the primary infection. The finding of mixed pairs of the two isolates indicates that worms of S. japonicum are either polygamous or able to wait in solitude for up to 12 weeks for a partner.