Strongyloides callosciureus n. sp. (Nematoda: Rhabditoidea), from Asian sciurids, is described based on morphology, morphometry, and the small and large subunit (SSU/LSU) ribosomal RNA gene (rDNA) sequences. This new species was collected from Pallas's squirrels (Callosciurus erythraeus) in the central part of mainland Japan (Honshu), which were originally introduced from Taiwan some decades ago, and plantain squirrels (Callosciurus notatus) imported from Malaysia as personal pets. For comparison, Strongyloides robustus Chandler, 1942 was collected from American red squirrels (Tamiasciurus hudsonicus) and southern flying squirrels (Glaucomys volans) imported from the United States as personal pets. The parasitic females found in North American and Asian sciurids shared some key morphological features such as the ovary running spirally around the gut, and the shapes of the stoma in the apical view and the tail. However, morphometric features of parasitic females in North American and Asian sciurids differed significantly from each other; the former was larger than the latter, and the relative position of the vulva to the whole body length from the mouth was different. The SSU/LSU rDNA sequences supported the division of sciurid Strongyloides isolates by geographical distribution of the host and morphological features, leading us to propose the erection of new species.
During 3 collecting expeditions between October 1996 and December 1996, fecal samples were obtained from 43 adult Gonocephalus grandis from Tanah Rata and the Cameron Highlands in Peninsular Malaysia. Two species of coccidia (Isospora gonocephali n. sp. [9/43, 23%] and Eimeria cameronensis n. sp. [3/43, 7%]) were discovered. Sporulated oocysts of I. gonocephali are subspherical to ovoidal, 22.3 x 18.7 (19-25 x 17-23) microm with a bilayered wall composed of a thin inner wall and a striated outer wall with a pitted surface; oocyst residuum absent; 1 polar granule present; sporocysts are almond-shaped, 13.5 x 9.2 (12-15 x 8.5-10) microm, Stieda body broad, domelike, substieda body fanlike, sporocyst residuum consisting of coarse, nonuniform granules in an amorphous cluster; sporozoites sausage-shaped with 1 large terminal, refractile body and lay randomly in the sporocyst. Sporulated oocysts of E. cameronensis are bilayered, smooth-walled, ellipsoidal, 26.5 x 12.4 (25-28 x 12-13) microm; with 1, small, polar granule composed of 2-3 splinter-like structures fused together; oocyst residuum absent; sporocysts ovoidal, almost rectangular-shaped 8.8 x 6.6 (8-9 x 5-7) microm, with no Stieda or substieda bodies, containing scattered residuum and 2 sausage-shaped sporozoites with 1 terminal, ovoidal refractile body. No individual lizard was host to both coccidian species.
Plasmodium inui (Halberstaedter and von Prowazek, 1907), a malarial parasite of Old World monkeys that occurs in isolated pockets throughout the Celebes, Indonesia, Malaysia, and the Philippines, has traditionally been considered to be related more closely to Plasmodium malariae of humans (and its primate counterpart Plasmodium brasilianum), than to other primate Plasmodium species. This inference was made in part because of the similarities in the periodicities or duration of the asexual cycle in the blood, the extended sporogonic cycle, and the longer period of time for development of the pre-erythrocytic stages in the liver. Both P. inui and P. malariae have quartan (72 hr) periodicities associated with their asexual cycle, whereas other primate malarias, such as Plasmodium fragile and Plasmodium cynomolgi, are associated with tertian periodicities (48 hr), and Plasmodiumn knowlesi, with a quotidian (24 hr) periodicity. Phylogenetic analyses of portions of orthologous small subunit ribosomal genes reveal that P. inui is actually more closely related to the Plasmodium species of the "vivax-type" lineage than to P. malariae. Ribosomal sequence analysis of many different, geographically isolated, antigenically distinct P. inui isolates reveals that the isolates are nearly identical in sequence and thus members of the same species.
Six clones were derived from each Malaysian Plasmodium falciparum isolate and characterized for their susceptibilities against type II antifolate drugs, cycloguanil and pyrimethamine. Results showed that these isolates were of a heterogeneous population, with average IC50 values of Gombak C clones at 0.012-0.084 microM and 0.027-0.066 microM, ST 9 clones at 0.019-0.258 microM and 0.027-0.241 microM, ST 12 clones at 0.015-0.342 microM and 0.012-0.107 microM, ST 85 clones at 0.022-0.087 microM and 0.024-0.426 microM, and ST 148 clones at 0.027-0312 microM and 0.029-0.690 microM against cycloguanil and pyrimethamine, respectively. Generally, most of these clones displayed susceptibility patterns similar to their parent isolates except ST 9/A4, ST 9/A7, ST 9/B5, ST 9/D9, ST 9/D10, ST 148/A4, ST 148/A5, ST 148/A7, ST 148/F7, ST 148/F8 clones, which were sensitive at 0.027 microM, 0.019 microM, 0.022 microM, 0.063 microM, 0.037 microM, 0.031 microM, 0.042, microM, 0.042 microM, 0.062 microM, and 0.027 microM, whereas, ST 12/D7 clone was resistant at 0.342 microM, against cycloguanil respectively. However, ST 9/A4, ST 9/D8, ST 12/D5, ST 85/A5, ST 85/B3, ST 85/B4, ST 85/D3, ST 85/D7, ST 148/A6, and ST 148/A7 clones were resistant to pyrimethamine at 0.158 microM, 0.241 microM, 0.107 microM, 0.223 microM, 0.393 microM, 0.402 microM, 0.426 microM, 0.115 microM, 0.690 microM, and 0.520 microM, respectively.
Schistosoma sinensium Pao, 1959, was first isolated from an unidentified snail in Szechuan Province, China, and was described on the basis of adult worms and eggs recovered from experimentally infected mice. We discovered snails, Tricula bollingi Davis, and rodents, Rattus rattus (L.) and Crocidura attenuata Milne-Edwards, infected with S. sinensium near Fang in northwest Thailand. Adult worms were recovered from small veins immediately adjacent to the small intestine. Eggs first appeared in the feces of experimentally infected mice at 25 days postinfection. Cercariae first emerged from experimentally infected T. bollingi at 40 days postinfection. Shortly after emerging, cercariae were found at the water surface, their bodies flat against the meniscus and tails hanging free in the water column. Our morphometric results for adults and eggs from naturally infected hosts agree well with those presented in the original description. The occurrence of a lateral spine on the egg of this Asian schistosome has created some confusion as to the affinities of S. sinensium. A comparison of 8 character states, including snail hosts, cercarial behavior, geographical distribution, and morphology of life cycle stages, indicates that S. sinensium is more closely related to S. japonicum Katsurada than it is to S. mansoni Sambon.
Moniliformis tarsii n. sp. was found in Tarsius bancanus. It is unique in possessing 11-12 longitudinal rows of 6-7 hooks each. Hooks 2 and 3 are conspicuously enlarged, 41-55 microns long. Moniliformis echinosorexi n. sp. differs from all other species in having 12-15 rows of 11-13 hooks that are 34-36 microns long, and in having a proboscis receptacle 1.2-2.0 mm long. Several new host records for M. moniliformis are presented.
Nanomedicine has the potential in enhancing the efficacy and bioavailability of anti-infective agents. Here we determined whether conjugation of the Malaysian cultivated seaweed Kappaphycus alvarezii with silver-conjugated nanoparticles enhanced anti-acanthamoebic properties. Silver-conjugated K. alvarezii were successfully synthesized, followed by characterization with Fourier transform infrared spectroscopy, ultraviolet-visible spectrophotometry, and transmission electron microscopy. Amoebicidal effects were evaluated against Acanthamoeba castellanii, and cytotoxicity assays were performed using HaCaT cells. Viability assays revealed that silver nanoparticles conjugated with K. alvarezii extract exhibited significant antiamoebic properties (P < 0.05). Nano-conjugates induced the production of reactive oxygen species. Importantly, silver-conjugated extract inhibited amoeba-mediated host cell damage as established by lactate dehydrogenase release. Neither the nano-conjugates nor the extract showed cytotoxicity against human cells in vitro. Liquid chromatography and mass spectroscopy revealed several molecules, including 2,6-nonadien-1-ol, N-desmethyl trifluoperazine, dulciol B, lucidumol A, acetoxolone, 2-[4,6-bis(2,4-dimethylphenyl)-1,3,5-triazin-2-yl]-5-(octyloxy)phenol, C16 sphinganine, 22-tricosenoic acid, and β-dihydrorotenone, of which dulciol B and C16 sphinganine are known to possess antimicrobial activities. In summary, marine organisms are an important source of bioactive molecules with anti-acanthamoebic properties that can be enhanced by conjugating with silver nanoparticles. Natural products combined with nanotechnology using multifunctional nanoparticle complexes can deliver therapeutic agents effectively and hold promise in the development of new formulations of anti-acanthamoebic agents.
Developmental and morphological characteristics of 3 isolates of Taenia taeniaeformis isolated from Clethrionomys rufocanus bedfordiae in Abuta (70 km southwest of Sapporo), Japan (isolate ACR), and from Rattus norvegicus in Sapporo, Japan (isolate SRN) and Kuala Lumpur, Malaysia (isolate KRN) were compared. Eggs of 3 isolates were administered to several species of rodents. Isolate ACR infected C. rufocanus bedfordiae, Apodemus speciosus, and Apodemus argenteus, but not rats or mice, whereas isolate SRN and isolate KRN were infective to rats, mice, A. speciosus, and A. argenteus, but not to C. rufocanus bedfordiae. The increase in cyst size of isolate ACR continued during the experimental period, whereas that of the other 2 isolates had ceased growing after 30 days postinfection. However, significant differences were observed in the length of the small rostellar hooks, number and distribution of testes, and the length of the cirrus sac between isolate ACR and the other 2 isolates. Thus it is suggested that isolate ACR is a distinct strain or even a new species.
A new species of parasitic nematode, Paracapillaria malayensis n. sp. (Capillariidae), is described from the small intestine of the toad Duttaphrynus melanostictus imported from the Malayan Peninsula to the Czech Republic. The new species differs from the only other congeneric species, Paracapillaria spratti, mainly in the shape and structure of the spicular proximal end (with a lobular rim), smaller eggs (45-51 x 21-24 microm), longer spicule (336 microm), and the number (37-38) of stichocytes in gravid females; whereas P. spratti parasitizes frogs of the Microhylidae in Papua New Guinea, P. malayensis is a parasite of Bufonidae in the Malayan Peninsula. Other Paracapillaria spp. are parasites of fishes, birds, or mammals and they mostly differ from P. malayensis in the structure of eggs and some other morphological features.