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.     

Evaluation of the infectivity of Trichinella spp. for reptiles (Caiman sclerops)

Experimental inoculation with nine well-characterised Trichinella isolates was performed on caimans (Caiman sclerops) to determine their infectivity for reptiles belonging to the family Crocodilidae. As controls, the same larval batches of Trichinella isolates were inoculated into mice and guinea pigs. It was suggested that Trichinella pseudospiralis was more likely to infect reptiles than encapsulating species, but whereas all Trichinella species established in mice and guinea pigs, the caimans remained negative. The finding that caimans could not be experimentally infected contrasts with a recent report on infections in farmed crocodiles (Crocodylus niloticus).     

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.     

The genetic analysis of F1 hybrid larvae between female Trichinella spiralis and male Trichinella britovi

Hybrids between female Trichinella spiralis and male Trichinella britovi were constructed. Then, hybrid genotype was characterized by DNA markers including mitochondrial cytochrome c oxidase subunit I (CO I) gene, the gene encoding the 43-kDa excretory–secretory (ES) protein, and genomic DNA fragments specific for T. spiralis and T. britovi identified from random amplified polymorphism DNA (RAPD). PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial CO I gene revealed that all hybrids carried a T. spiralis pattern. The same analysis of the gene encoding the 43-kDa ES protein showed that each hybrid carried both T. spiralis and T. britovi gene type simultaneously. In the analysis of genomic DNA using RAPD-derived PCR primers, some hybrids carried T. spiralis and T. britovi-specific RAPD markers, while others carried the RAPD marker of T. spiralis only.     

NADH dehydrogenase subunit 1 and cytochrome c oxidase subunit I sequences compared for members of the genus Taenia (Cestoda)

Nine members of the genus Taenia (Taenia taeniaeformis, Taenia hydatigena, Taenia pisiformis, Taenia ovis, Taenia multiceps, Taenia serialis, Taenia saginata, Taenia solium and the Asian Taenia) were characterised by their mitochondrial NADH dehydrogenase subunit 1 gene sequences and their genetic relationships were compared with those derived from the cytochrome c oxidase subunit I sequence data. The extent of inter-taxon sequence difference in NADH dehydrogenase subunit 1 (5.9–30.8%) was usually greater than in cytochrome c oxidase subunit I (2.5–18%). Although topology of the phenograms derived from NADH dehydrogenase subunit 1 and cytochrome c oxidase subunit I sequence data differed, there was concordance in that T. multiceps, T. serialis (of canids), T. saginata and the Asian Taenia (of humans) were genetically most similar, and those four members were genetically more similar to T. ovis and T. solium than they were to T. hydatigena and T. pisiformis (of canids) or T. taeniaeformis (of cats). The NADH dehydrogenase subunit 1 sequence data may prove useful in studies of the systematics and population genetic structure of the Taeniidae.     

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.     

Effect of acute versus chronic Trichinella pseudospiralis infections on systemic cell-mediated immunity.

Infection of the mouse with Trichinella pseudospiralis is accompanied by pronounced suppression of host inflammatory response. This study examines the effects of infection with this parasite on several key elements in cell-mediated immunity. Early down-regulation of host granulomatous response to subcutaneously implanted cotton string and delayed-type hypersensitivity (DTH) response to trinitrochlorobenzene (TNCB) was followed later during infection by normalization of these parameters compared to that seen in uninfected mice. Cytotoxic T lymphocyte responses to tumor-specific antigens expressed on the syngeneic P91 mastocytoma were depressed early following infection with T. pseudospiralis relative to that seen in uninfected mice but were similar in these two groups during the later stages of infection. Down-regulation of the components of cell-mediated reaction examined herein accompanied the presence of migratory larvae in the host.     

Nouvelles données sur les estérases des trichogrammes (hym. trichogrammatidae)

New electrophoretic analysis allowed us to show that the two fastest esterases of T. brassicae are derived from the Est 5′ locus and not from the Est 5 and Est 6 locus. This result changes only slightly the evanescens group place in the phylogenetic tree previously suggested. Esterases have been studied in several other species. Those of T. lacustre and T. piceum are not against the classification of these species in the perkinsi and fasciatum groups, respectively. Those of T. leptoparameron lead us to place this species in the pretiosum group, near T. daumalae. Those of a population of T. dendrolimi reveal a polymorphism clearly higher than what was known in bisexual forms of this species. Lastly, esterases analysed by other authors tend to show that T. chilonis and T. closterae belong to the nubilale group (the appellation of which has to be changed) and that T. ostriniae belongs to the minutum group.     

Trichinella papuae n.sp. (Nematoda), a new non-encapsulated species from domestic and sylvatic swine of Papua New Guinea

Encapsulated and non-encapsulated species of the genus Trichinella are widespread in sylvatic animals in almost all zoogeographical regions. In sylvatic animals from Tasmania (Australian region), only the non-encapsulated species Trichinella pseudospiralis has been reported. Between 1988 and 1998, non-encapsulated larvae of Trichinella were detected in five domestic pigs and six wild boars from a remote area of Papua New Guinea. Morphological, biological, and molecular studies carried out on one strain isolated from a wild boar in 1997 suggest that these parasites belong to a new species, which has been named Trichinella papuae n.sp. This species can be identified by the morphology of muscle larvae, which lack a nurse cell in host muscles, and whose total length is one-third greater than that of the other non-encapsulated species, T. pseudospiralis. Adults of T. papuae do not cross with adults of the other species and genotypes. Muscle larvae of T. papuae are unable to infect birds, whereas those of T. pseudospiralis do. The expansion segment V of the large subunit of the ribosomal DNA differs from that of the other species and genotypes. All of these features allow for the easy identification of T. papuae, even in poorly equipped laboratories. The discovery and identification of a second non-encapsulated species in the Australian region strongly supports the existence of two evolutionary lines in the genus Trichinella, which differ in terms of the capacity of larvae to induce a modification of the muscle cell into a nurse cell.     

Cryopreservation of the first-stage larvae of trichostrongylid nematode parasites

First stage (L1) larvae of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta can be cryopreserved in the presence of DMSO using a two-step freezing protocol involving an initial period at −80°C prior to transfer to liquid nitrogen. Thawed L1 larvae continue development in vitro producing third stage (L3) larvae that are infective to sheep when dosed per os. Establishment rates for L3 larvae grown from thawed L1 larvae were 40 and 80% for H. contortus and T. colubriformis, respectively. There was no difference in survival or infectivity between benzimidazole (BZ)-susceptible and BZ-resistant H. contortus isolates and cryopreservation caused no shift in their BZ-resistance status as indicated in an in vitro larval development assay. Cryopreservation also had no effect on the sensitivity of these isolates to the avermectins or levamisole in vitro. High survival rates (60–70%), good levels of establishment and the stability of anthelmintic resistance status of isolates indicate that little if any selection occurs during the cryopreservation process. L1 larvae of all 3 species have been successfully recovered after 16 months storage in liquid nitrogen, cultured to the L3 stage and established in sheep.     

Sequence variation in the cytochrome oxidase I, internal transcribed spacer 1, and Ts14 diagnostic antigen sequences of Taenia solium isolates from South and Central America, India, and Asia.

We examined the genetic variability in the pig–human tapeworm, Taenia solium, by sequencing the genes for cytochrome oxidase I, internal transcribed spacer 1, and a diagnostic antigen, Ts14, from individual cysts isolated from Peru, Colombia, Mexico, India, China, and the Philippines. For these genes, the rate of nucleotide variation was minimal. Isolates from these countries can be distinguished based on one to eight nucleotide differences in the 396 nucleotide cytochrome oxidase I (COI) sequence. However, all of the 15 isolates from within Peru had identical COI sequences. The Ts14 sequences from India and China were identical and differed from the Peru sequence by three nucleotides in 333. These data indicate that there is minimal genetic variability within the species T. solium. Minimal variability was also seen in the ITS1 sequence, but this variation was observed within the individual. Twenty-two cloned sequences from six isolates sorted into 13 unique sequences. The variability observed within the sequences from individual cysts was as great as the variability between the isolates.     

The refractory responses of the White Swiss strain of Mus musculus to infection with Taenia taeniaeformis

Penetration of intestinal mucosa by oncospheres of Taenia taeniaeformis occurred among three age groups of White Swiss Mus musculus. Highest frequency of penetration was demonstrated among the 10–30 day-old group with the other two older groups, 40–60 and 70–100 days, each having succeedingly less penetration. The invasion process was more cytolytic than ground substance lysis; no difference in this process, or the reaction of the villi to it, was detected among the various age groups. Liver reaction, displayed by the same three age groups against larvae of T. taeniaeformis, was age dependent. Disorganization, loss of larval tissue integrity, and formation of a non-cellular capsule were demonstrated within 48 h in the 10–30 day group, while a rapid cellular response of inflammation and walling-off of larvae was seen in the 40–60 day group. No larvae were observed in the 70–100 day group.     

The use of specific lysine modifications to locate the reaction site of cytochrome c with sulfite oxidase

The reduction of cytochrome c by beef liver sulfite oxidase was found to be strongly inhibited by high ionic strength, indicating the importance of electrostatic interactions to the reaction. The reaction rates of sulfite oxidase with singly trifluoroacetylated or trifluoromethylphenylcarbamylated cytochrome c derivatives were studied to determine the role of individual lysines in the reaction. The reaction rate was decreased by modification of the lysines immediately surrounding the heme crevice, the decreases following the order: Lys 13 > Lys 25 Lys 79 ≈ Lys 87 > Lys 8 ≈ Lys 27 ≈ Lys 72. Modification of lysines 22, 55, 88, 99, and 100 had no effect on the reaction rate. These results indicate that the interaction site on cytochrome c for sulfite oxidase is at the heme crevice region, and overlaps considerable with that for cytochrome oxidase.     

Laboratory culture of the Nudibranch Tritonia diomedea bergh (Tritoniidae: Opisthobranchia) and some aspects of its behavioral development

Tritonia diomedea Bergh was reared from oviposition, through metamorphosis to reproductive maturity in the laboratory. The larvae of T. diomedea are planktotrophic and undergo considerable shell growth (from 144.6–329 μ average maximum shell length). Metamorphosis does not require induction, but there may be a preference to metamorphose in the presence of the probable adult prey, a small Virgularia sp. Larvae in cultures fed no food, Dunaliella tertiolecta Butcher Isochrysis galbana Parke, or Monochrysis lutheri Droop did not achieve metamorphic competence at near ambient sea-water temperatures (11.9±1.3 and 13.0±0.8°C). Larvae from cultures fed Monochrysis at room temperature (20.8±1.5°C) or fed a 1 : 1 mixture of Isochrysis and Monochrysis at near ambient sea-water temperatures did metamorphose. Even so, only those larvae fed the 1 : 1 mixture survived more than a few days following metamorphosis. Adult behavioral patterns developed gradually, feeding being first observed at 5 days, swimming in response to NaCl crystals at about 60 days, copulation at about 272 days, and oviposition at about 277 days after metamorphosis. Growth rates were determined for field collected Tritonia diomedea; smaller animals gained and lost weight relatively faster than larger animals.     

Iron protein content of Thiocapsa pfennigii, a purple sulfur bacterium of atypical chlorophyll composition

Four out of five soluble electron transport iron proteins of Thiocapsa pfennigii plus the particulate cytochromes have been found to be analogous to those of Chromatium vinosum strain D. In addition to ferredoxin, high potential iron-sulfur protein. cytochrome c′, and cytochrome c-552(550), T. pfennigii contains a cytochrome c-552(545) not previously isolated from photosynthetic bacteria. It is concluded that T. pfennigii is more closely related to C. vinosum than to Rhodopseudomonas viridis, the only other known bacterial species having bacteriochlorophyll b.     

环颈蝠(Thainycteris aureocollaris)在中国分布的再发现

福建省位于我国东南沿海,亚热带气候和丰富植被为野生动物的生存和繁衍提供了良好的生态环境(丁晖等,2015).武夷山位于福建北部,地理位置独特,雨量充足,植被多样且茂密,是众多动物赖以生存的场所,丰富的生物多样性资源使其成为世界著名的动物模式标本产地(罗桂环等,2016;耿宝荣等,2020),其中翼手目中模式产地为武夷山...     

Synergism and antagonism between two trematode species in the snail Lymnaea rubiginosa

Synergism and antagonism between two trematode species in the snail Lymnaea rubiginosa. Internationaljournal for Parasitology 3: 729–733. Sporocysts of Trichobilharzia brevis in the snail exerted a synergistic effect on sporocysts of Echinostoma hystricosum: The rate of infection with E. hystricosum was much higher in snails harboring T. brevis than in control snails with no other infection. Rediae of E. hystricosum and sporocysts of T. brevis were antagonistic, the predatory rediae consuming the sporocysts and ultimately eliminating T. brevis from the snail. Once a snail was occupied by E. hystricosum it could not be superinfected by T. brevis.     

山西省南方红豆杉自然分布与群落生态学特征

南方红豆杉为国家Ⅰ级重点保护植物,山西省东南部为其自然分布最北界.本文调查了南方红豆杉在山西省的野生分布,对其进行了群落划分、物种多样性、结构特征和竞争特性的研究.结果表明: 山西省南方红豆杉主要形成南方红豆杉+鹅耳枥群落、南方红豆杉-荆条群落、南方红豆杉-海州常山群落和南方红豆杉+栓皮栎群落,集中分布于陵川县磨河和阳城县蟒河自然保护区,其他地区均为散生;群落垂直结构明显,南方红豆杉已进入主林层,混交林林冠层高8~10 m,纯林高5～6 m;群落间的总体物种多样性指数和均匀度指数存在显著差异.南方红豆杉植株矮小,平均高5.16 m,乔木层、演替层分别占43.4%和56.6%,更新层缺乏;胸径＜16 cm的个体占67.6%,胸径32～40 cm的个体占4.0%,幼苗稀少,仅发现4株幼苗;大量的小径级个体虽可保证南方红豆杉在一定时间内维持稳定的种群结构,但幼苗缺失必将引起未来种群的衰退.山西省野生南方红豆杉分布相对集中,多数是小径级个体,导致其种内竞争激烈,种内竞争强度占61.8%;鹅耳枥和栓皮栎是研究地区的主要组成树种,对南方红豆杉造成的种间竞争压力最大.     

Epidemiological studies on horses infected with nematodes of the family Trichonematidae (Witenberg, 1925)

Observations are reported on seasonal changes in the age structure of populations of nematodes of the family Trichonematidae recovered at regular intervals from the lumens of the large intestines of horses slaughtered in S.W. England. The results show that changes in the size of parasitic populations of Trichonema nassatum follow seasonal variations in the rate of infection, more individuals maturing during summer/autumn than during winter/spring because of proportional differences in the numbers of infective larvae ingested from the pasture. In contrast, larvae of T. longibursatum, T. catinatum and T. goldi ingested by grazing horses during summer accumulate in the gut wall rather than promptly returning to the lumen and developing to maturity. Their development is apparently inhibited until the following spring when 4th-stage larvae emerge en masse and quickly reach the adult stage. The relationship between fluctuations in the size of the adult populations of the 4 species studied and the characteristic seasonal variations in horse faecal egg counts is discussed.     

On the embryology and mode of reproduction of selected diploid species of Hieracium s.l. (Asteraceae) from Bulgaria

Three diploid taxa – Hieracium transylvanicum (subgenus Hieracium), Hieracium caespitosum subsp. brevipilum and Hieracium pavichii (subgenus Pilosella) – from five natural Bulgarian populations were investigated embryologically. The peculiarities of the male and female gametophytes, embryo- and endospermogenesis were established in each species. The results suggest that the species propagate sexually as expected from their diploid chromosome number. However, some forms of apomixis have also been observed, e.g. somatic apospory and integumental embryony in H. pavichii. The presence of apomixis in a diploid taxon shows that polyploidy is not an obligatory prerequisite for apomixis and the two phenomena are independent. The embryological plasticity detected in H. pavichii and H. transylvanicum suggests they may have higher opportunities for adaptation and successful reproduction.