Molecular investigation of Anaplasma species in sheep from Heilongjiang Province,northeast China identified four Anaplasma species and a novel genotype of Anaplasma capra

Anaplasmosis poses a great threat to the livestock industry and human health in most tropical and subtropical regions of the world. This study investigated the presence of Anaplasma in sheep from Heilongjiang Province, northeastern China. A total of 341 blood samples were detected by PCR with species-specific primers based on the msp4 gene of Anaplasma ovis, 16S rRNA gene of Anaplasma phagocytophilum and Anaplasma bovis and gltA gene of Anaplasma capra. The results showed that Anaplasma infection was found in 103 (30.2%) of 341 sheep. The infection rates were 2.6%, 8.8%, 15.8% and 10.0% for A. ovis, A. phagocytophilum, A. bovis and A. capra in sheep, respectively. Co-infection involving two Anaplasma species was found in 25 sheep (8.0%), which were usually A. phagocytophilum and A. bovis (72.0%). Co-infection involving A. phagocytophilum, A. capra, A. ovis with zoonotic potential, was found in one sheep. Sequence analysis revealed that the isolates of A. ovis, A. bovis and A. phagocytophilum identified in sheep were closely related to those previously reported in ticks and other animal hosts. Phylogenetic analysis showed that A. capra could be classified into two distinct clusters based on the gltA gene and the isolates identified in sheep from this study were clustered in the A. capra genotype II, which was clearly distinct with the human isolates. The findings in this study report four Anaplasma species and a novel A. capra genotype in sheep from northeastern China, and improve our knowledge of Anaplasma, contributing to the control of ovine anaplasmosis.     

Molecular survey of Anaplasma phagocytophilum and Ehrlichia canis in red foxes (Vulpes vulpes) from central Italy

During the 2007-2008 hunting season, 150 spleen samples were collected from free-ranging red foxes (Vulpes vulpes) in central Italy. The specimens were tested by two nested PCR assays to detect DNA of Anaplasma phagocytophilum, etiologic agent of granulocytic ehrlichiosis of animals and humans, and DNA of Ehrlichia canis, which causes the monocytic ehrlichiosis in canids. None of the foxes were PCR-positive for E. canis; 25 (16.6%) were positive for A. phagocytophilum. No specific gross alterations were detected at necropsy, and no histopathologic lesions found on PCR-positive spleen samples.     

渤海中、南部大型底栖动物物种多样性的研究

以1997年 6月、1998年9月和1999年4月共3个航次,覆盖渤海中、南部大部分海区的大型底栖动物丰度、生物量和生产力的研究为基础,进一步对其物种多样性进行了分析。结果表明：研究海域的大型底栖动物物种多样性没有明显的平面分布规律;物种多样性指数与沉积物因子的相关性高于其与底层水因子的相关性;沉积物中污染物含量的确会对大型底栖动物的多样性造成影响。通过对3个航次中3个相同站位的比较,发现不同年份之3个季节的平均Shannon Wiener指数无显著差异（P>0.05）;而1998年9月和1999年4月2个航次中20个相同站位的平均Shannon Wiener指数同样也未表现出明显的差异（P>0.05）。此外,渤海的物种数量要明显地高于胶州湾。     

Molecular identification of mosquito species in the Anopheles annularis group in southern Asia

The Anopheles annularis group of subgenus Cellia Theobald (Diptera: Culicidae) includes five currently recognized species in southern Asia: An. annularis Van der Wulp, Anopheles nivipes (Theobald) and Anopheles philippinensis Ludlow, which are widespread in the region, Anopheles pallidus Theobald, which is known in Sri Lanka, India and Myanmar, and Anopheles schueffneri Stanton, which occurs in Java and Sumatra. Identification of the four mainland species based on morphology is problematic. In view of the fact that the three widespread species are variously involved in malaria transmission in different parts of the region, we developed a species-specific polymerase chain reaction assay based on rDNA internal transcribed spacer 2 (ITS2) sequences to facilitate entomological and epidemiological studies of the four species. The method proved to be reliable when tested over a wide geographical area.     

Molecular genetic identification of southern hemisphere beaked whales (Cetacea: Ziphiidae)

To assist in the species-level identification of stranded and hunted beaked whales, we compiled a database of 'reference' sequences from the mitochondrial DNA control region for 15 of the 20 described ziphiid species. Reference samples for eight species were obtained from stranded animals in New Zealand and South Australia. Sequences for a further seven species were obtained from a previously published report. This database was used to identify 20 'test' samples obtained from incompletely documented strandings around New Zealand. Analyses showed that four of these 'test' specimens (20%) had initially been misidentified. These included two animals of particular interest: (i) a Blainville's beaked whale ( Mesoplodon densirostris) , the first record of this species in New Zealand waters; and, (ii) a juvenile Andrews' beaked whale ( Mesoplodon bowdoini ), a species known from just over 20 strandings worldwide. A published sequence from a beaked whale product purchased in the Republic of Korea was identified as a Cuvier's beaked whale ( Ziphius cavirostris ). Levels of intra- and interspecific variation were compared to determine the potential for misidentification when the database or taxonomy is incomplete. Intraspecific variation was generally <2%, and interspecific divergence was generally >4.7%. Exceptions were within-species variation in Hyperoodon planifrons , southern bottlenosed whale (4.12%), which exceeded the variation between the two species of Berardius (3.78%), and variation between the two specimens assigned to M. hectori , Hector's beaked whale (7.14%). The latter case appears to be an error in species identification, and could represent the discovery of a new species of beaked whale.     

Molecular identification and genetic diversity within species of the genera Hanseniaspora and Kloeckera

Three molecular methods, RAPD-PCR analysis, electrophoretic karyotyping and RFLP of the PCR-amplified ITS regions (ITS1, ITS2 and the intervening 5.8S rDNA), were studied for accurate identification of Hanseniaspora and Kloeckera species as well as for determining inter- and intraspecific relationships of 74 strains isolated from different sources and/or geographically distinct regions. Of these three methods, PCR-RFLP analysis of ITS regions with restriction enzymes DdeI and HinfI is proposed as a rapid identification method to discriminate unambiguously between all six Hanseniaspora species and the single non-ascospore-forming apiculate yeast species Kloeckera lindneri. Electrophoretic karyotyping produced chromosomal profiles by which the seven species could be divided into four groups sharing similar karyotypes. Although most of the 60 strains examined exhibited a common species-specific pattern, a different degree of chromosomal-length polymorphism and a variable number of chromosomal DNA fragments were observed within species. Cluster analysis of the combined RAPD-PCR fingerprints obtained with one 10-mer primer, two microsatellite primers and one minisatellite primer generated clusters which with a few exceptions are in agreement with the groups as earlier recognized in DNA-DNA homology studies.     

五种常见嗜尸性蝇类的分子鉴定

为了解决法医学人员对于嗜尸性蝇类的鉴定难题,对中山市、广州市及西安市5种常见嗜尸性蝇类共17个样本,对其线粒体COⅠ基因的348 bp大小片段进行了RFLP和DNA序列分析,采用ABI377测序仪测序,DNASTAR软件预测限制性位点,DdeⅠ, DraⅠ和HinfⅠ3种限制性内切酶消化,非变性聚丙烯酰胺凝胶电泳检测酶切结果,MEGA3.0软件包进行序列分析和构建系统发育树。结果表明： 采用mtDNA扩增结合银染技术和DNA序列分析,均可以方便快捷地进行上述三地5种常见嗜尸性蝇类的种类鉴定。本文结果为法医昆虫学中嗜尸性昆虫DNA鉴定数据库的建立提供了数据资料。     

Molecular phylogeography and genetic diversity of East Asian goats

The domestic goat is one of the most important livestock species, but its origins and genetic diversity still remain uncertain. Multiple highly divergent maternal lineages of goat have been reported in previous studies. Although one of the mitochondrial DNA lineages, lineage B, was detected only in eastern and southern Asia, the geographic distribution of these lineages was previously unclear. Here, we examine the genetic diversity and phylogeographic structure of Asian goats by mitochondrial DNA sequences and morphological characteristics. The analyses of a total of 1661 Asian goats from 12 countries revealed a high frequency of lineage B in Southeast Asia. The frequency of this lineage tended to be higher in mountain areas than in plain areas in Southeast Asian countries, and there was a significant correlation between its frequency and morphological traits. The results suggest an original predominance of lineage B in Southeast Asia and the recent infiltration of lineage A into Southeast Asian goats.     

Two new species of Taeniolina from southern China

Taeniolina euryae sp. nov. and T. schimae sp. nov. were discovered on dead branches of Eurya macartneyi and Schima superba, respectively, in tropical forests of southern China. They are described, illustrated and compared with closely related taxa. A key to species of Taeniolina is provided.     

Three new species of Sporidesmiella from southern China

Sporidesmiella rosae sp. nov., S. machili sp. nov., and S. archidendri sp. nov., occurring, respectively, on dead branches of Rosa chinensis, Machilus grijsii, and Archidendron clypearia, were collected from subtropical forests in southern China. They are described, illustrated, and compared with similar taxa. A dichotomous key to Sporidesmiella species is provided.     

中国南方地区暗色丝孢真菌两个新种（英文）

霸王岭爱氏霉和版纳葚孢是采自我国南方地区的暗色丝孢菌的2个新种。霸王岭爱氏霉的主要特征是分生孢子梗分化明显,单生,产孢细胞单芽生并产生17–24个假隔膜,115–145×15.5–20μm,倒棍棒形的分生孢子。版纳葚孢的主要特征是分生孢子梗粗大,单生,产孢细胞单芽生并产生11–25个真隔膜,215–430×12–16μm,倒棍棒形分生孢子。     

Molecular survey of Hepatozoon species in lizards from North Africa

The prevalence of Hepatozoon parasites in 460 lizards from North Africa was studied by amplification and sequencing of the 18S rRNA gene. The phylogenetic analysis of the 18S rRNA gene provides new insights into the phylogeny of these parasites with multiple genetically distinct lineages recovered. Parasite prevalence differed significantly between lacertid lizards and geckos. Our results show that there is limited host specificity and no clear relation to the geographical distribution of Hepatozoon parasites.     

Molecular identification and genotyping of Blastocystis sp. in sheep and goats from some areas in Inner Mongolia,Northern China

Blastocystis sp. is a kind of unicellular intestinal commensal which is widely distributed in humans and animals, and frequently found in the people who are in close contact with animals. To investigate the prevalence and evaluate the zoonotic potential of Blastocystis sp. in sheep and goats from Inner Mongolia, China, a total of 1037 samples were collected from them, and subjected to nested PCR amplification based on the small subunit ribosomal RNA (SSU rRNA) gene of Blastocystis sp. The sanger sequencing was used for Blastocystis sp. subtype identification. The results indicated that the average infection rate of Blastocystis sp. was 10.70% [95CI: 8.82%–12.58%] (111/1037), including 11.30% [95CI: 7.96%–14.64%] for sheep (39/345) and 10.40% [95CI: 8.13%–12.67%] for goats (72/692). Five Blastocystis subtypes (ST5, ST10, ST14, ST21 and ST26) were identified in the present study. Among them, ST10 was the most dominant subtype in sheep and goats, accounting for 70.27% (78/111) of the total identified positive samples. This is the first report regarding Blastocystis sp. subtypes ST21 and ST26 in goats in China. This study has provided a detail epidemiological data on the prevalence and subtypes distribution of Blastocystis sp. in sheep and goats in Inner Mongolia, China. Our results indicated that sheep and goats could be reservoir host for multiple Bastocystis subtypes, including the zoonotic subtypes. Further studies among humans, livestock and wild animals are needed to better understand their role in the spread of Blastocystis sp.     

The development of PCR methodology for the identification of species of the tapeworm Moniezia from cattle, goats and sheep in central Vietnam

The aims of this study were to investigate the prevalence of Moniezia spp. in domestic ruminants in central Vietnam and to develop a polymerase chain reaction (PCR) technique to distinguish M. expansa from M. benedeni. Among 2040 examined domestic animals (540 cattle, 800 goats, 700 sheep) Moniezia was recovered from 5.4% of cattle, 16.4% of sheep and 20.6% of goats. A set of primers for PCR was designed to classify M. expansa and M. benedeni based on the amplification of DNA corresponding to the internal transcribed spacer of 5.8S rRNA. The 457 specimens (75 from cattle, 162 from goats, 150 from sheep, 30 from horses, 30 from chickens and 10 from dogs) were subjected to PCR for classification of Moniezia spp. PCR products with the expected sizes were amplified from bovine, ovine and caprine specimens. No specific PCR products were found for specimens from horses, chickens and dogs. Of the 75 specimens from cattle, nine were classified as M. expansa and 66 were M. benedeni. Among 162 caprine specimens, 138 were M. expansa and 24 were M. benedeni. The distribution of M. expansa and M. benedeni in 150 ovine specimens was 132 and 18, respectively. These results show that M. expansa is dominant in goats and sheep, whereas M. benedeni is more common in cattle; PCR can be used for classification of these two species.     

Molecular genetic variation in Passiflora alata (Passifloraceae), an invasive species in southern Brazil

A total of 85 wild plants were collected in different areas of southern and south-eastern Brazil, and studied for nrDNA internal transcribed spacers (ITS). Thirty-two of them were also investigated for two other nuclear ( G3pdh and LEAFY ), as well as eight plastid systems. Sixty-one ITS sequence types were identified, in a region of 412 bp, whereas 42 G3pdh haplotypes were found in a region of 1010 bp, and 23 LEAFY haplotypes in a region of 565 bp. GC content was higher in ITS (65%) than in the two other nuclear regions (40–42%), whereas nucleotide diversity was markedly higher in G3pdh than in the other two. No variability was found in the plastid systems. Most of the nuclear molecular diversity (56–86%) occurs at the intrapopulation level, and there is clear evidence of population expansion and little geographical structure. Historical information and field studies indicate that Passiflora alata is actively colonizing previously unoccupied areas in southern Brazil. High indices of intrapopulation variability were found in the region that is being colonized, similar to those found in areas where P. alata is native. Multiple introductions, as well the species' genetic structure and human interference, may explain these findings.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 611–630.     

Isolation and identification of Acanthamoeba species from thermal spring environments in southern Taiwan

Acanthamoeba species are free-living amoebae found in a range of environments. Within this genus, a number of species are recognized as human pathogens, potentially causing Acanthamoeba keratitis, granulomatous amoebic encephalitis, and chronic granulomatous lesions. In this study, 60 water samples were taken from four thermal spring recreation areas in southern Taiwan. We detected living Acanthamoeba spp. based on culture-confirmed detection combined with the molecular taxonomic identification method. Living Acanthamoeba spp. were detected in nine (15%) samples. The presence or absence of Acanthamoeba spp. in the water samples depended significantly on the pH value. The most frequently identified living Acanthamoeba genotype was T15 followed by T4, Acanthamoeba spp., and T2. Genotypes T2, T4, and T15 of Acanthamoeba, are responsible for Acanthamoeba keratitis as well as granulomatous amoebic encephalitis, and should therefore be considered a potential health risk associated with human activities in thermal spring environments.     

Ethnobotanical survey of cooling herbal drinks from southern China

In terms of scientific activities generally and ethnobiological pursuits in particular, North Korea, officially known as the Democratic People’s Republic of Korea, is an almost blank entity on the quilt of global research. During a sabbatical semester at Pyongyang University of Science and Technology the author used this opportunity to gather some information on the uses of insect and other terrestrial arthropods as human food and components of traditional healing methods in that country. Despite the widely publicised shortcomings in the supply of food stuffs to the population of North Korea, insects are not generally seen as a source of food worthy of exploitation. However, the therapeutic use of insects, centipedes and scorpions to treat illnesses as diverse as the common cold, skin rashes, constipation, dysentery, nervous prostration, whooping cough, osteomyelitis, tetanus, and various forms of cancer is apparently still popular. The arthropods used therapeutically are credited with anti-inflammatory, immunological and other health-promoting effects, because they are said to contain hormones, steroids, lipids and plant-derived alkaloids, all of which capable of exerting their effects on the human body.     

Molecular identification of Candida species from urinary infections in Honduras

Background

Candiduria is a common infection among hospitalised patients. Although the clinical relevance of yeasts in urine is not clearly defined, fungal urinary tract infections have increased significantly in the last decades, becoming a growing public health problem. Candida albicans is the most commonly reported species in urinary infections, although other species of the genus are becoming particularly important, because some of them are linked with resistance to antifungal drugs.

Molecular identification of Pilobolus species from Yellowstone National Park

Pilobolus, a widely distributed coprophilous fungus, grows on herbivore dung. Species of Pilobolus traditionally are described with imprecise morphological characteristics potentially leading to misidentification. In this study we used PCR analysis of taxonomically informative sequences to provide more consistent species identification from isolates obtained in Yellowstone National Park. We collected Pilobolus park-wide from six taxa of herbivores over 9 y. Multiple transfers of single sporangium isolates provided pure cultures from which DNA was extracted. Sequence analysis of internal transcribed spacer (ITS) regions of DNA that code for rRNA genes were used to distinguish among similar species. We describe several species of Pilobolus associated with herbivores in various habitats, including two species not previously reported, P. heterosporus and P. sphaerosporus. Our results show that phylogenetic species identification of Pilobolus based on sequence analysis of pure culture isolates provides a more reliable means of identifying species than traditional methods.     

Morphological and molecular identification of two new species of Tubulicrinis (Hymenochaetaceae,Hymenochaetales) from southern China

Two new wood-inhabiting fungal species, Tubulicrinis xantha and T. yunnanensis spp. nov., are described based on morphological and molecular characters. Tubulicrinis xantha is characterized by resupinate, furfuraceous basidiomata with buff to yellowish hymenial surface, amyloid lyocystidia and cylindrical to allantoid, hyaline, thin-walled, smooth basidiospores (5.3–6.3 × 1.2–1.6 μm). Tubulicrinis yunnanensis is characterized by pruinose basidiomata with primrose to olivaceous hymenial surface and encrusted lyocystidia and cylindrical, hyaline, thin-walled, smooth basidiospores (4.2–6.2 × 1.2–2 μm). Sequences of ITS and LSU nrRNA gene regions of the studied samples were generated, and phylogenetic analyses were performed with maximum likelihood, maximum parsimony and bayesian inference methods. The phylogenetic analyses based on molecular data of ITS and ITS + nLSU sequences showed that T. xantha grouped with T. martinicensis, and T. yunnanensis grouped with T. glebulosus.