An IC-PCR method for detection of Cryptosporidium and Giardia in natural surface waters in Finland

We developed an immunocapture-based polymerase chain reaction (PCR) assay for simultaneous detection of Cryptosporidium parvum oocysts and Giardia intestinalis cysts in surface water. Using primer pairs Cry9/Cry15 and LaxA/LaxB for Cryptosporidium and Gdh1/Gdh4 for Giardia, the sensitivity of the entire detection procedure (dealing with concentration, separation, DNA purification and PCR amplification) was at the level of 50–100 oocysts and cysts. Of 54 surface water samples, 4 were positive for Cryptosporidium and 1 for Giardia. Cryptosporidium and Giardia were detected for the first time in surface water in Finland.     

Prevalence and genotyping of human isolates of Giardia duodenalis from Albania

Microscopical and PCR-based techniques were performed in order to investigate the prevalence of infection and the genotypes of Giardia duodenalis from 125 stool samples collected from children living in the urban and the rural areas of Tirana (Albania) and hospitalized with acute gastroenteritis. 7 out of 125 samples resulted positive for Giardia at the microscopic examination (5.6%). In 50 selected samples including the 7 samples positive for Giardia by microscopy, 3 and 15 additional positive samples were detected by immunofluorescence and PCR, respectively. Seasonality appeared as an important parameter to be evaluated in order to better understand the prevalence of infection. Sequence analysis revealed both human Assemblage A and B. This result represents the first data on G. duodenalis genotypes in Albania.     

Relationships between four novel ceratomyxid parasites from the gall bladders of labrid fishes from Heron Island, Queensland, Australia

Ceratomyxid parasites from the gall bladder of fishes from the family Labridae were examined for their taxonomic identity and their relatedness to other species of myxozoans. This paper describes four novel species Ceratomyxa choerodonae n. sp. from Choerodon cyanodus; Ceratomyxa cheilinae n. sp. from Cheilinus chlorourus; Ceratomyxa oxycheilinae n. sp. from Oxycheilinus digramma; and Ceratomyxa thalassomae n. sp. from Thalassoma lunare. Each species is characterised morphologically and small subunit (18 S) rDNA sequences were used in molecular phylogenetic analyses. Comparison of the small subunit rDNA sequences revealed four distinct species even though morphological differences indicated the presence of only two morphologically distinctive spore types. Phylogenetic analyses revealed that the ceratomyxids from labrid fishes did not form a clade to the exclusion of all other myxosporeans and provides evidence that host relatedness is not a good character to discriminate between species within the genus Ceratomyxa.     

A retrospective study of the microbiological and parasitological status of laboratory rodents in France

Between 1988 and 1997, 72 mouse colonies and 38 rat colonies were examined for the presence of bacteria parasite infections. Among mouse and rat bacteria, high positive rates were observed with Proteus species (sp.), Pasteurella pneumotropica, Mycoplasma sp. and Pseudomonas aeruginosa. Concerning murine colonies, parasites frequently detected were Tritrichomonas sp., Syphacia sp., Aspiculuris tetraptera, Entamoeba muris, Spironucleus muris, Myobia musculi, Chilomastix sp. and Myocoptes musculinus. In rats, high rates were obtained with Syphacia sp., Tritrichomonas sp., Spironucleus muris, Entamoeba muris and Chilomastix sp. During the first part of the last decade, some agents such as Clostridium piliforme, Citrobacter sp., Mycoplasma sp., Pseudomonas aeruginosa, Myobia musculi, Radfordia ensifera, Spironucleus muris and Giardia muris were often found among rodents, and most of them were still present in 1997. At the time of our study, results point out that some agents are still persistent, even increasing during the same period. It is particularly the case for parasites such as Entamoeba muris and the oxyurids, but also for bacteria like Proteus sp. and Klebsiella pneumoniae. We can thus conclude that only very limited success has been achieved in preventing microbial and parasitic infections in mice and rats colonies.     

冬季上海市城区河道中原生动物群落的结构特征

于2007年11月至2008年1月,对上海市城区经过初步修复的四条河道(午潮港、横港、朝阳河和曹杨环浜)中原生动物的群落结构和多样性进行了调查.共鉴定出原生动物88种,其中鞭毛虫31种,肉足虫13种,纤毛虫44种.除曹杨环浜无优势种外,其他三条河道的种类均以耐污种类为优势种,如波豆虫 Bodo sp.、滴虫 Monas sp.、隐藻 Cryptomonas sp.等;四条河道均呈现纤毛虫和鞭毛虫种类较为丰富,肉足虫相对较少的现象.四条河道中原生动物多样性指数的平均值变动在1.14～2.87之间;密度变动在900.35～2837.89 ind/ml间.分析发现,前期生态治理较为成功的曹杨环浜具有多样性指数高、密度大、无明显优势种等特征,而其他三条河道中原生动物种类相对较单一,且分布不均匀,主要优势种为一些耐污种类.提示经过人工合理的修复,可以有效改善受污染河道的状况.因此对已经初步修复河道的生态系统进行评估,探索和制定出有针对性的修复方案,并对其加以合理的维护,对建立健康、稳态的河道生态系统是十分必要的.     

Implications of the new eukaryotic systematics for parasitologists

An accurate understanding of evolutionary relationships is central in biology. For parasitologists, understanding the relationships among eukaryotic organisms allows the prediction of virulence mechanisms, reconstruction of metabolic pathways, identification of potential drug targets, elucidation of parasite-specific cellular processes and understanding of interactions with the host or vector. Here we consider the impact of major recent revisions of eukaryotic systematics and taxonomy on parasitology. The previous, ladder-like model placed some protists as early diverging, with the remaining eukaryotes “progressing” towards a “crown radiation” of animals, plants, Fungi and some additional protistan lineages. This model has been robustly disproven. The new model is based on vastly increased amounts of molecular sequence data, integration with morphological information and the rigorous application of phylogenetic methods to those data. It now divides eukaryotes into six major supergroups; the relationships between those groups and the order of branching remain unknown. This new eukaryotic phylogeny emphasizes that organisms including Giardia, Trypanosoma and Trichomonas are not primitive, but instead highly evolved and specialised for their specific environments. The wealth of newly available comparative genomic data has also allowed the reconstruction of ancient suites of characteristics and mapping of character evolution in diverse parasites. For example, the last common eukaryotic ancestor was apparently complex, suggesting that lineage-specific adaptations and secondary losses have been important in the evolution of protistan parasites. Referring to the best evidence-based models for eukaryotic evolution will allow parasitologists to make more accurate and reliable inferences about pathogens that cause significant morbidity and mortality.     

陕西圈养珍稀野生动物肠道寄生虫感染及其形态观察

2007年11～12月对陕西省珍稀野生动物抢救饲养研究中心中的珍稀野生动物进行了寄生虫感染情况及其种类、形态的观察.采用生理盐水涂片法、碘液染色法对18种75头/只野生动物的粪便进行检查,对检出的寄生虫进行数码显微摄片.结果共检出11种寄生虫,总感染率为88.9%,以芽囊原虫(Blaaocyais sp.)、阿米巴原虫感染较为突出.     

Hematozoa in endemic subspecies of common kestrel in the Cape Verde Islands

We examined 130 Common Kestrel (Falco tinnunculus) representing two endemic subspecies and nine resident island populations on the Cape Verde archipelago between 1996 and 1999 to study diversity, prevalence, and intensity of hematozoa. Hematozoan diversity was very low; we detected only Plasmodium fallax, a species that is rarely found in Falconoformes, and, possibly, Haemoproteus brachiatus. Moreover, prevalence of Plasmodium fallax was low (1.5%) with a mean intensity of infection of 0.05 protozoa/10(-3) erythrocytes. Only one bird (0.8%) was infected with a gametocyte that was most likely Haemoproteus brachiatus; the intensity in this infected bird was 1.5 protozoa/10(-3) erythrocytes. A single parasite or two parasites were observed in blood smears in four additional birds, but identification to genus was not possible. This is the first record of blood parasites in birds on the Cape Verde Archipelago. The low prevalence of these parasites might be because of arid and less-favorable conditions for the pathogen's vectors. The sedentary nature and high level of isolation of the island kestrel populations are also factors that could decrease the probability of infection.     

Experimental aerosol inoculation of Mycobacterium bovis in North American opossums (Didelphis virginiana)

The goal of this study was to evaluate the susceptibility of North American opossums (Didelphis virginiana) to aerosol inoculation of Mycobacterium bovis at two dose levels in order to gain information on disease pathogenesis, fecal shedding of the organism, and the potential role that opossums play in the spread of this disease in nature. Six opossums received high dose (1 x 10(7) colony forming units (cfu) by aerosol inoculation, six opossums received low dose (1 x 10(3) cfu inoculation, and six opossums were sham-inoculated with sterile water and served as controls. Lungs were the most frequently infected tissues, with nine of 12 inoculated opossums positive for M. bovis on culture. Gross lesions consisted of multifocal pneumonia and enlarged lymph nodes. Microscopically, granulomatous pneumonia and granulomatous lymphadenitis associated with acid-fast bacilli were present in eight of 12 inoculated opossums. Fecal shedding of M. bovis was uncommon at both inoculation doses. While opossums were highly susceptible to aerosol inoculation of M. bovis, they did not become emaciated or develop widely disseminated lesions. From this study, opossums may transmit tuberculosis by aerosol infection to other opossums in close contact and serve as a source of infection to carnivores that feed upon them, however, transmission of the disease to large herbivores by fecal shedding or direct contact may be less likely.     

Coprozoic Protozoa from Wyoming Mammals

SYNOPSIS. Coprozoic protozoa from elk, bison, bear, moose, coyote, marmot, cattle, horse, sheep and man were kept in feces at 4° C. for 2–6 months. During that time in most of the samples there developed the flagellates Cercomonas sp., Copromonas ruminantium and Monas communis; the amoebae Vahlkampfia sp. and Sappinia diploidea , and ciliates of the Nyctotherus type, and an unidentified smaller species. There was a correlation between numbers of protozoa and bacteria. The same species of protozoa in soil or in soil mixed with boiled feces failed to live. Coprozoic protozoa may require certain essential metabolites from bacteria as do true parasites. The wide variation in appearance of cysts made it practically impossible to identify the protozoa with certainty in that stage. Reliance had to be on motile forms which readily developed in the cold cultures.     

Hematozoa of raptors from southern New Jersey and adjacent areas

Blood smears from 259 birds of 12 species, representing four families of raptors, from New Jersey, Pennsylvania, Delaware, and Virginia were examined for blood parasites. Infected birds constituted 59.1% of the total. Birds were infected with one or more of the following genera of protozoa: Leucocytozoon (43.2%); Haemoproteus (21.6%); Plasmodium (1.2%); and Trypanosoma (1.2%). Blood culture of 142 raptors of 11 species for Trypanosoma revealed a prevalence of 41.5%. Plasmodium circumflexum is reported for the first time in Accipiter striatus, and Trypanosoma sp. in Buteo jamaicensis.     

Status of intestinal parasites infection among primary school children in Kampongcham,Cambodia

A survey was made to find the extent of intestinal parasite infection in Kampongcham, Cambodia in February 2002. A total of 251 fecal specimens were collected from Tonlebat primary school children and examined by formalin-ether sedimentation technique. The overall infection rate of intestinal parasite was 54.2% (males, 57.3%; females, 50.8%). The infection rate of intestinal helminths by the species were as follows: Ascaris lumbricoides 26.3%, Echinostoma sp. 15.6%, hookworm 6.4%, Opisthorchis sp. 4.0%, Rhabditis sp. 2.4%, and Trichuris trichiura 0.4%. The infection rate of intestinal protozoa were as follows: E. coli 7.6%, G. lamblia 3.2%, I. butschlii 3.2%, and E. histolytica 0.8%. More than two different kinds of parasites were found in 16.7% of the stool samples. All the children infected were treated with albendazole, praziquantel and metronidazole according to parasite species. The results showed that intestinal parasites are highly endemic in this area.     

内蒙古天竺葵腐霉枯萎病病原菌

为了明确天竺葵腐霉枯萎病的病原菌种类,从内蒙古通辽采集天竺葵腐霉枯萎病样品,对病原物进行了分离、纯化和致病性测定,并对病原物进行形态学和rDNA-ITS序列分析鉴定。结果表明,从10个植物样品中分离得到16个腐霉菌株,分别属于终极腐霉Pythium ultimum var. ultimum、瓜果腐霉P. aphanidermatum和两个与P. ultimum var. ultimum相似的待定种。其中,P. ultimum var. ultimum的分离频率为75%、P. aphanidermatum的分离频率为12.5%,两个腐霉待定种的分离频率均为6.25%;P. ultimum var. ultimum是优势类群。致病性测定结果表明,4种腐霉菌都能引起天竺葵腐霉枯萎病,与自然发病症状相同,其中P. ultimum var. ultimum和P. aphanidermatum的致病性较强,发病率分别为71.4%和85.7%。     

Chemical composition of essential oils of seven species of Eugenia from Monteverde, Costa Rica

The leaf essential oils of seven species of Eugenia from Monteverde, Costa Rica (Eugenia austin-smithii, Eugenia cartagensis, Eugenia haberi, Eugenia monteverdensis, Eugenia zuchowskiae, Eugenia sp. A aff. haberi, and Eugenia sp. B aff. oerstediana) have been obtained by hydrodistillation and analyzed by GC–MS. The seven species were compared to determine the similarities and differences among their volatile chemical compositions. The major component in each of the seven species was as follows: E. austin-smithii and E. cartagensis was trans-2-hexenal, E. haberi and E. zuchowskiae was -pinene, E. monteverdensis was linalool, Eugenia sp. A was zingiberene, and Eugenia sp. B was 1,8-cineole. The following six components were present in all seven species: -copaene, β-caryophyllene, -humulene, δ-cadinene, trans-nerolidol, and torreyol. The complex array and differing abundances of these compounds among the Eugenia species studied suggest that they may provide useful characters in understanding the phylogenetic relationships among closely related species.     

Three new intestinal protozoan species of the genus Latteuria n.g. (Ciliophora: Trichostomatia) from Asian and African elephants

Three new ciliate species presumably belonging to the family Paraisotrichidae were recognized in the fecal samples from zoo-kept Asian and African elephants. All the ciliates possess a unique but similar arrangement of somatic ciliature, thus a new genus Latteuria n.g. has been created for them. The genus is characterized by the presence of a tapered frontal ‘spout’ at the anterior end of the body, posterior ciliary rows in narrow grooves encircling the posterior half of the body and an anterior arch of cilia. Latteuria polyfaria n.sp. is the largest species in the genus with 9–11 posterior ciliary rows. In L. media n.sp., of medium body size, the number of rows varies from four to six, and the smallest species, L. trifaria n.sp., has only three-four posterior ciliary rows.     

黑竹冲河小裳蜉(Leptophlebia sp.)和蜉蝣(Ephemera sp.)周年生产量

大型底栖动物在河流生态系统中发挥着重要作用。2003年6月至2004年6月间对汉江流域2级河流--黑竹冲河大型底栖动物群落优势种类的生产力进行了为期一周年的调查研究。结果表明,主要优势种小裳蜉(Leptophlebia sp.)和蜉蝣(Ephemera sp.)的生活史均为一年两代,蜉蝣的两个代之间界限比较明显;小裳蜉成虫羽化主要发生在秋季和冬季,而蜉蝣成虫羽化主要发生在夏季和冬季。小裳蜉和蜉蝣种群的生物量和多度均出现两次峰值。采用体长频率法(size-frequencymethod)测算的周年生产量分别为:小裳蜉,38.0362g·m^-2·a^-1,P/B为11.4;蜉蝣,76.0318g·m^-2·a^-1,P/B为11.8。     

Status of intestinal parasite infections among children in Bat Dambang, Cambodia

A survey was conducted to determine the extent of intestinal parasite infection in Bat Dambang, Cambodia in March 2004. A total of 623 fecal specimens was collected from kindergarten and schoolchildren and examined using the formalin-ether sedimentation technique. The overall infection rate of intestinal parasites was 25.7% (boys, 26.2%; girls, 25.1%), and the infection rates of intestinal helminthes by species were as follows: Echinostoma sp. 4.8%, hookworm 3.4%, Hymenolepis nana 1.3%, and Rhabditis sp. 1.3%. The infection rates of intestinal protozoa were; Entamoeba coli 4.8%, Giardia lamblia 2.9%, Iodamoeba butschlii 1.4%, Entamoeba polecki 1.1%, and Entamoeba histolytica 0.8%. There were no egg positive cases of Ascaris lumbricoides or Trichuris trichiura. All children infected were treated with albendazole, praziquantel, or metronidazole according to parasite species. The results showed that intestinal parasites are highly endemic in Bat Dambang, Cambodia.     

Intracellular parasitism: cell biological adaptations of parasitic protozoa to a life inside cells

Several protozoan parasites evade the host's immune defence because most of their development takes place inside specific host cells. Only a few of these protozoa live within the host cell cytosol. Most parasites are sequestered within membrane-bound compartments, collectively called ‘vacuoles’. Recent advances in the cell biology of intracellular parasites have revealed fundamental differences in the strategies whereby such organisms gain entry into their respective host cells. These differences have important implications for host-parasite interaction and for nutrient acquisition by the parasite. Leishmania spp. take advantage of the phagocytic properties of their host cells and presumably contribute little to the uptake process. In contrast, apicomplexan parasites have developed highly specialised organelles, called micronemes and rhoptries, to actively invade a variety of nucleated cells and, in the case of Plasmodium falciparum, human erythrocytes. Following invasion, parasites use a multitude of strategies to protect themselves from the defence mechanisms of the parasitized cells. In addition, they induce novel pathways within the infected cell that allow a most efficient nutrient acquisition both from the host cell cytoplasm and from the extracellular environment. Parasite-induced changes of host cells are most apparent in erythrocytes infected with Plasmodium spp. Mammalian erythrocytes are deficient in de novo protein and lipid biosynthesis and, consequently, pathways which allow the transport of macromolecules and small solutes are established by metabolic activities of the parasite. Research into the cell biology of intracellular parasitism has identified fascinating phenomena some of which we are beginning to understand at a molecular level. They are fascinating because they allow insights into a very intimate interaction between two eukaryotic cells of entirely different phylogenetic origins.     

Host adaptation and host-parasite co-evolution in Cryptosporidium: implications for taxonomy and public health

To assess the genetic diversity and evolution of Cryptosporidium parasites, the partial ssrRNA, actin, and 70 kDa heat shock protein (HSP70) genes of 15 new Cryptosporidium parasites were sequenced. Sequence data were analysed together with those previously obtained from other Cryptosporidium parasites (10 Cryptosporidium spp. and eight Cryptosporidium genotypes). Results of this multi-locus genetic characterisation indicate that host adaptation is a general phenomenon in the genus Cryptosporidium, because specific genotypes were usually associated with specific groups of animals. On the other hand, host–parasite co-evolution is also common in Cryptosporidium, as closely related hosts usually had related Cryptosporidium parasites. Results of phylogenetic analyses suggest that the Cryptosporidium parvum bovine genotype and Cryptosporidium meleagridis were originally parasites of rodents and mammals, respectively, but have subsequently expanded their host ranges to include humans. Understanding the evolution of Cryptosporidium species is important not only for clarification of the taxonomy of the parasites but also for assessment of the public health significance of Cryptosporidium parasites from animals.