Clinostomum attenuatum (Digenea) from the eye of Bufo marinus

Two specimens of the cane toad, Bufo marinus, were found with single worm infestations of the eye. One worm was excysted in the eye chamber, the other still encysted in the choroid layer; both worms were identified as Clinostomum attenuatum. This finding is both a new host record and a new site of development for this amphibian parasite.     

Clinostomum complanatum and Clinostomum marginatum (Rudolphi, 1819) (Digenea: Clinostomidae) are separate species based on differences in ribosomal DNA

Infections by metacercariae of Clinostomum (Leidy, 1856) species adversely affect aquacultured fish and are potentially transmissible to humans. Molecular methodologies are efficient tools, which enable diagnosis of all life-history stages of trematodes in their diverse hosts. The small subunit of ribosomal DNA genes of adults of the Old World Clinostomum complanatum (Rudolphi, 1819) and the New World Clinostomum marginatum (Rudolphi, 1819), obtained from a little egret Egretta garzetta (Linnaeus, 1766) and the great blue heron Ardea herodias (Linnaeus, 1758), respectively, were amplified, sequenced, and aligned. The resulting alignment was used to develop a genetic assay to differentiate between these species.     

Parasitism of Cichlasoma paranaense Kullander, 1983 and Gymnotus carapo Linnaeus, 1814 by Clinostomum complanatum (Rudolphi, 1814) metacercariae in the Taquari river.

The infection, known as yellow spot disease, produced by metacercariae of Clinostomum spp. was studied in fishes of the Taquari river, located in Jataizinho, Paraná State, Brazil. A total of 1,582 specimens, belonging to 36 species, were collected between March 1999 and April 2001. Yellow spot disease was observed only in Gymnotus carapo Linnaeus, 1814 (Gymnotiformes, Gymnotidae) and Cichlasoma paranaense Kullander, 1983 (Perciformes, Cichlidae). This parasitism was generated by metacercariae of Clinostomum complanatum (Rudolphi, 1814) (Digenea, Clinostomidae). Among the 88 specimens of G. carapo examined, 7 (prevalence = 8%) had cysts of the parasite. Four of them were captured in July 1999 and three in October 1999. In the other months, no specimen found was infected. Using relative condition factor (Kn) analysis, it was determined that both infected and non-infected specimens had a total weight equal to the theoretically expected value for each total length (Kn = 1.0). Among 56 individuals of Cichlasoma paranaense, 6 (prevalence = 10.7%) had between 1 and 27 metacercariae of C. complanatum (mean intensity of infection = 9.3 +/- 9.6). In March 1999 and April of both 2000 and 2001, the specimens examined were not infected. The infected fish had a total weight higher than the expected value (Kn > 1.0), while the non-infected fish had a weight equal to the expected value (Kn = 1.0).     

Digenetic trematodes of the genus Clinostomum Leidy, 1856 (Digenea: Clinostomidae) from birds of Queensland, Australia, including C. wilsoni n. sp. from Egretta intermedia

Two species of Clinostomum previously described from Australia, C. hornum from Botaurus poiciloptilus (Australian bittern) and Nycticorax caledonicus (Nankeen night heron) and C. australiense from Pelecanus conspicillatus (Australian pelican), which have previously been synonymised with C. complanatum, are redescribed and recognised as valid species. In addition, C. complanatum is recorded from Egretta alba (large egret), E. garzetta (little egret), E. intermedia (plumed egret), N. caledonicus and Ardea novaehollandiae (white-faced heron). C. wilsoni n. sp. is described from E. intermedia from Queensland. C. wilsoni differs from the other three species in size and shape of the body and in the oral collar, oral sucker, intestinal caeca, caecal diverticula and position of testes. Taxonomic problems within the genus Clinostomum are discussed.     

Distribution, abundance, and genetic diversity of Clinostomum spp. metacercariae (Trematoda:Digenea) in a modified Ozark stream system

Land-use alterations can have profound influences on faunal distributions, including host-parasite relationships. Yellow grub trematodes ( Clinostomum spp.) have complex life cycles involving 3 hosts: a snail, a fish or amphibian, and a bird. Here, we analyze the distribution, prevalence, intensity, abundance, and genetic diversity of encysting metacercariae of Clinostomum spp. in salamanders and fishes throughout an aquatic system that includes a natural Ozark stream and man-made ponds. We found Clinostomum sp. infecting permanently aquatic Oklahoma salamanders ( Eurycea tynerensis ; 56% prevalence) and larval grotto salamanders ( Eurycea spelaea ) immediately downstream from a man-made pond. However, Clinostomum sp. did not infect any salamanders in the spring that supplies this pond, or in sections farther downstream (~0.5 and 2 km). Metacercariae of Clinostomum sp. were present in ~90% of introduced largemouth bass ( Micropterus salmoides ) in the man-made pond adjunct to the stream. Morphological examination and phylogenetic analyses based on the mitochondrial gene cytochrome oxidase 1 ( Co1 ) and the nuclear ribosomal gene 18S show that fishes and salamanders at this site are primarily infected with Clinostomum marginatum . There is a relatively high degree of mitochondrial haplotype diversity in C. marginatum at this site but no consistent genetic difference between parasites in largemouth bass from the man-made pond and those in salamanders from the stream. Based on the microgeographic distribution and relationships of metacercariae of C. marginatum at this site, we hypothesize that the adjunct man-made pond has created an ecological situation that brings the cercariae of this parasite into contact with novel stream salamander hosts.     

Study of the Chemical Constituents in Hypericum attenuatum Chois

Four crystalline components had been isolated from Hypericum attenuatum Chins., which is wild plant in Hulun Buir Meng of Nei Monggol Zizhiqu. On the basis of spectral analysis, hydrolyzed acid, the prepared derivatives, and the pysico-chemical constants crystalline Ⅰ, Ⅱ of components had been identified as hyperin (hyperoside) and quercetin respectively, crystalline Ⅲ identified with chromatograph is chlorogenic acid with a few impurity. The component Ⅳ is a flavonoid compound too, but it is not identified. The average of the contents of flavonoid in the whole herb is 1.67%, but the contents of flavonoids in the different organs as the flower, the fruit, the leaf and the stem are quite different. The content in the leaf is highest, that in the reproductive organs is secondary and that in the stem is lowest. The resources of Hypericum attenuatum Chois., are abundant in nature. The content of flavonoids in Hypericum attenuatum Chois. is higher. Its pharmacological function is extensive. Therefore, it is a kind of medicinal herb worthily for further study.     

Biologically active substances isolated from Hypericum attenuatum Choisy

Four dianthrones, two oxymethylanthraquinones, two reductive forms of anthraquinones and five flavonoids have been isolated from the plant source Hypericum attenuatum Choisy (Guttiferae) firstly investigated for these substances contents. Dianthrones pseudohypericin, protohypericin, protopseudohypericin, oxymethylanthraquinones frangulaemodin, frangulin, reductive anthraquinones emodin anthrone, emodin anthranol, and flavonoids quercetin, quercitrin, isoquercitrin, rutin have been identified for the first time. High hyperoside contents in herb of Hypericum attenuatum is established.     

不同产地、不同物候期对乌腺金丝桃中总黄酮含量的影响

目的:探讨不同产地、不同物候期乌腺金丝桃中总黄酮含量的变化。方法:采用比色法测定了3个产地乌腺金丝桃在不同物候期总黄酮的含量。以总黄酮为技术指标筛选乌腺金丝桃药材的采收期、黄酮含量最高的产地。结果:以总黄酮成分为指标确定的产地、采收期为:佳木斯猴石山产,花盛期采收。结论:通过研究发现乌腺金丝桃药材以勃利县产花盛期采收总黄酮含量最高,对乌腺金丝桃的开发有一定指导意义。     

Morphological and molecular differentiation of Clinostomum complanatum and Clinostomum marginatum (Digenea: Clinostomidae) metacercariae and adults

The separation of Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819 has long been unclear. Recent data confirm the validity of the junior species, C. marginatum , by ～ 1% differences in its 18S rDNA sequences. We collected adults and metacercariae of C. complanatum and C. marginatum and found reliable morphological differences in the genital complex at both developmental stages. In addition, we identified basic morphometrics (distance between suckers, body width) in metacercariae that may be useful for discriminating the species. The morphological differences were supported by the comparison of sequences of internal transcribed spacers of ribosomal DNA and of the mitochondrial gene cytochrome c oxidase I (COI) from 39 specimens. In 36 specimens, the average divergence between the species was 7.3% in ITS and 19.4% in COI sequences. Two specimens from North America and 1 from Europe had sequences that did not allow them to be clearly allied with either species.     

Clinostomum complanatum (Trematoda: Clinostomatidae) in five new fish hosts in Japan.

Metacercariae of Clinostomum complanatum were found in six species of natural freshwater fishes collected at Koyama Pond in Tottori City of Tottori Prefecture, Japan. Five of these fish species are reported here as new records for second intermediate hosts of C. complanatum in Japan: the silver crucian carp (Carassius gibelio langsdorfi), the deepbodied crucian carp (Carassius cuvieri), the carp (Cyprinus carpio), the topmouth gudgeon (Pseudorasbora parva) and the rose bitterling (Rhodeus ocelatus).     

渐狭蜡蚧菌产生的几丁质酶对芦笋茎枯病菌的抑制作用

目的:研究渐狭蜡蚧菌产生的几丁质酶及其对芦笋茎枯病菌天门冬拟茎点霉的抑制效果。方法:通过乙酰葡萄糖胺法、对硝基苯酚法和活性电泳法测定渐狭蜡蚧菌CGMCC5328的产几丁质酶特性及其粗酶液对芦笋茎枯病菌的抑菌效果。结果:渐狭蜡蚧菌CGMCC5328几丁质酶粗提取液的几丁质降解酶系活性第4 d达到高峰期;几丁质外切酶活性第6 d达到高峰期;活性染色检测到5条几丁质酶谱带,相对分子质量分别为32.9×103、42.1×103、54.1×103、65.6×103和79.6×103。用该几丁质酶粗酶液处理芦笋茎枯病菌,与灭活酶液处理相比较,抑菌效果明显,酶液处理导致菌丝畸形。结论:渐狭蜡蚧菌CGMCC5328产生的几丁质酶对芦笋茎枯病菌有抑制作用。     

Helminths from the fish Dormitator maculatus (Osteichthyes: Eleotridae) from Alvarado, Veracruz, Mexico

Dormirator maculatus (n=184) was collected in the Alvarado Lagoon, Mexico during a year period (Oct. 1993-1994). In the helminthologic review, the presence of Clinostomum complanatum (82.3%), Neoechinorhynchus golvani (76.1%), Spiroxys sp. (21.3 %), and Camallanus sp. (6.2%) was registered. Reduction of the hematocrit caused by such infection is significant (t, a= 0.05).     

The Effect of Activated Charcoal Supplemented Media to Browning of In Vitro Cultures of Piper species

With the aim to effectively minimise the browning of tissue cultures of different Piper species (P. longum, P. attenuatum, P. betle, P. nigrum) explants from stem (nodal and internodal segments), petiole, and leaf were planted on Murashige and Skoog's basal medium supplemented with activated charcoal (AC). AC in the concentration of 200 mg dm^-3 proved to be the best. Among the taxa studied, P. longum showed the least browning whereas, wild P. nigrum showed maximum browning.     

Parasites with zoonotic potential found in commercially important fish in Tamaulipas,Northeastern Mexico

Human population is exposed to numerous parasitic ichthyozoonoses. Although Tamaulipas state (northeastern Mexico) is well known for its fishing and aquaculture industry, there are few reports of this type of zoonosis. Therefore, it is imperative to investigate whether the parasites that affect these fish may represent a zoonotic risk for the inhabitants of the area.The objective of this study was to identify molecular and/or morphologically muscle parasites of fish from coastal locations in Tamaulipas, Mexico, and assess the risk of infection for humans. Between 2017 and 2018, 764 individual fish belonging to 28 species were examined for parasites. Collected worms were processed for their identification using morphological characteristics. In addition, partial sequences of the large subunit (28S) ribosomal RNA gene were obtained from some species to corroborate their identity. Prevalence and mean intensity of all registered infections were calculated. A total of seven species of parasites were found: cestodes (Poecilancistrium caryophyllum), trematodes (Clinostomum tataxumui, Clinostomum cichlidorum), nematodes (Eustrongylides sp., Contracaecum sp.) and pentastomids (Sebekia purdieae, Sebekia sp.). Parasites infected 10 species belonging to different fish families (Ariidae, Centrarchidae, Centropomidae, Cichlidae, Eleotridae, Ictaluridae, Mugilidae and Sciaenidae). Congeneric species of parasites or related to those registered in this study have been identified as zoonotic agents in other regions of the world. Despite the low levels of infection (2.6–16.6% prevalence and 1–5.5 parasites per infected host), there is a latent risk of transmission to humans, so it is recommended to avoid eating raw or undercooked fish meat.     

Natural infections of Clinostomum complanatum (Trematoda: Clinostomatidae) in wild herons and egrets, Tottori Prefecture, Japan.

Nycticorax nycticorax, Ardea cinerea, Egretta garzetta, and Egretta intermedia were naturally infected with Clinostomum complanatum (Trematoda: Clinostomatidae) among fourteen wild herons, seven wild egrets and one wild bittern evaluated at the Veterinary Hospital of Tottori University, Tottori, Japan. The latter three species of heron and egrets are reported for the first time as definitive hosts of this parasite in Japan.     

Mechanisms associated with cellular desiccation tolerance of Artemia encysted embryos from locations around the world

Using differential scanning calorimetry we demonstrated the presence of biological glasses and measured the glass transition temperatures (Tg) in dry encysted gastrula embryos (cysts) of the brine shrimp, Artemia, from eleven different locations, two of which provided cysts from parthenogenetic animals. Values for Tg were highest, by far, in Artemia franciscana cysts from the Mekong Delta, Vietnam (VN), these cysts having been produced from previous sequential inoculations into growth ponds of cysts from the San Francisco Bay, California, USA. Tg values for three groups of A. franciscana cysts were significantly higher than those of other cysts (except those of Artemia persimilis) studied here, as well as all other desiccation-tolerant animal systems studied to date. We also measured three stress proteins (hsc70, artemin and p26) in all these cysts as well as the total alcohol soluble carbohydrates (ASC), about 90% of which is the disaccharide trehalose, a known component of biological glasses. We interpret the results in terms of mechanisms involved with desiccation tolerance and, to some extent, with thermal conditions at the sites of cyst collection.     

Mechanisms associated with cellular desiccation tolerance in the animal extremophile artemia

Using differential scanning calorimetry, we demonstrated the presence of biological glasses and measured the transition temperatures in dry encysted embryos (cysts) of the brine shrimp, Artemia franciscana. Cysts from the following three geographic locations were studied: San Francisco Bay (SFB); the Great Salt Lake, Utah (GSL); and the Mekong Delta, Vietnam (VN; these cysts were produced from previous sequential inoculations of SFB cysts into growth ponds). Values for the glass transition temperature, T(g), were highest in VN cysts. This study indicates that the composition and properties of these biological glasses can be altered by natural selection and thermal adaptation. To our knowledge, T(g) values for all three kinds of cysts were significantly higher than those for any other desiccation-tolerant animal system. To gain insight into the significance of T(g), we examined the thermal stability of these dry cysts at 80 °C. GSL cysts were the least tolerant, by far, with VN cysts being extremely tolerant and SFB cysts not far behind. Those results correlated with the thermal transition values. Also measured were alcohol-soluble carbohydrates, ~90% of which is the disaccharide trehalose, a known component of biological glasses. Amounts in the GSL cysts were significantly less than those in the other two kinds of cysts. Several stress proteins were measured in the three groups of cysts, with all of them being in lesser amounts in GSL cysts compared with the SFB and VN cysts. We interpret the data in terms of mechanisms involved with desiccation tolerance and thermal conditions at the sites of cyst collection.     

Morphological variability and degenerative evolution of human hepatic hydatid cysts

The findings are presented of a macro and microscopic investigation of 89 hydatid hepatic cysts removed intact from 59 patients by total pericystectomy. Detailed analysis revealed significant morphostructural variability and cysts grouped into 10 types were characterized, providing useful clinical indications. Only 30 cysts resulted fertile (33.7%), probably due to mean age of sample; 7 of these were "classic" cysts, 1 "septated" and 22 "multivesicular" packed with daughter cysts (DC), of varying turgidity or collapsed. Among the remaining 59 sterile cysts, 52 were degenerated and classified as "hyperlaminated" cysts due to the presence of large convoluted sheets of laminar tissue (SLT) surrounded by varying amounts of caseous (40 specimens), granular (6) or gelatinous (6) matrix. Moreover, "multivesicular", "acephalocyst", "caseous" and "serous" cysts were also recovered among the sterile specimens. Some "multivesicular" cysts (14) appeared as "transitional forms" towards the various types of "hyperlaminated" cysts containing all different forms of DC and large SLT intermingled with a variously degenerated matrix. The comprehensive study allows to hypothesize the train of events leading, over the years, to the gradual transformation and degeneration of the larval form Echinococcus granulosus in the human liver.     

Cross-species transmission of Giardia spp.: inoculation of beavers and muskrats with cysts of human, beaver, mouse, and muskrat origin

Giardia cysts isolated from humans, beavers, mice, and muskrats were tested in cross-species transmission experiments for their ability to infect either beavers or muskrats. Giardia cysts, derived from multiple symptomatic human donors and used for inoculation of beavers or muskrats, were shown to be viable by incorporation of fluorogenic dyes, excystation, and their ability to produce infections in the Mongolian gerbil model. Inoculation of beavers with 5 x 10(5) Giardia lamblia cysts resulted in the infection of 75% of the animals (n = 8), as judged by the presence of fecal cysts or intestinal trophozoites at necropsy. The mean prepatent period was 13.1 days. An infective dose experiment, using 5 x 10(1) to 5 x 10(5) viable G. lamblia cysts collected by fluorescence-activated cell sorting, demonstrated that doses of between, less than 50, and less than 500 viable cysts were required to produce infection in beavers. Scanning electron microscopy of beaver small intestine revealed that attachment of G. lamblia trophozoites produced lesions in the microvillous border. Inoculation of muskrats with G. lamblia cysts produced infections when the dose of cysts was equal to or greater than 1.25 x 10(5). The inoculation of beavers with Giardia ondatrae or Giardia muris cysts did not produce any infection; however, the administration to muskrats of Giardia cysts of beaver origin resulted in the infection of 62% of the animals (n = 8), with a prepatent period of 5 days. Our results demonstrated that beavers and muskrats could be infected with Giardia cysts derived from humans, but only by using large numbers of cysts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Recovery of Cryptosporidium oocysts and Giardia cysts from source water concentrates using immunomagnetic separation

Immunomagnetic separation (IMS) procedures for the simultaneous isolation of Cryptosporidium oocysts and Giardia cysts have recently become available. We validated Dynal's GC-Combo IMS kit using source water at three turbidity levels (5000, 500 and 50 nephelometric turbidity units [ntu]) obtained from different geographical locations and spiked with approximately 9--11 (oo)cysts per ml. Mean recoveries of Cryptosporidium oocysts and Giardia cysts in deionized water were 62% and 69%, respectively. In turbid water matrices, mean recoveries of Cryptosporidium oocysts were between 55.9% and 83.1% while mean recoveries of cysts were between 61.1% and 89.6%. Marginally higher recoveries of the heat inactivated (oo)cysts were observed (119.4% Cryptosporidium oocysts and 90.9% Giardia cysts) in deionized water when compared with recoveries of viable (oo)cysts (69.7% Cryptosporidium oocysts and 79% Giardia cysts). Age of (oo)cysts on recoveries using the GC-Combo IMS kit demonstrated no effects up to 20 months old. Recovery of Giardia cysts was consistent for isolates aged up to 8 months (81.4%), however, a significant reduction in recoveries was noted at 20 months age. Recoveries of low levels (5 and 10 (oo)cysts) of Cryptosporidium oocysts and Giardia cysts in deionized water using IMS ranged from 51.3% to 78% and from 47.6% to 90.0%, respectively. Results of this study indicate that Dynal's GC-Combo IMS kit is an efficient technique to separate Cryptosporidium/Giardia from turbid matrices and yields consistent, reproducible recoveries. The use of fresh (recently voided and purified) (oo)cysts, aged (oo)cysts, viable and heat-inactivated (oo)cysts indicated that these parameters do not influence IMS performance.