Morphology and developmental rate of blowflies <Emphasis Type="Italic">Chrysomya megacephala</Emphasis> and <Emphasis Type="Italic">Chrysomya rufifacies</Emphasis> in Thailand: application in forensic entomology

The larval morphology and developmental rate of Chrysomya megacephala (F.) and Chrysomya rufifacies (Macquart), the two most forensically important blowfly species in Thailand, are presented. Morphological comparison of the third instar of both species revealed different characteristics (e.g., body appearance, cephalopharyngeal skeleton, dorsal cuticular spines between the prothorax and mesothorax, and feature of the posterior spiracle), thereby, allowing correct identification. A data analysis was conducted in Chiang Mai province, Northern Thailand during 2000–2001 on the developmental rate of both flies under natural ambient temperature and a natural light–dark photoperiod. The results indicated that larvae of C. megacephala developed more rapidly in April, pupariation initiated at 84 h at temperatures averaging 31.4°C, and the larvae grew slower in the rainy season and winter. Similarly, rapid development of C. rufifacies larvae appeared in the summer, with a pupariation period as short as 96 h in June (average temperature 27.4°C). Analysis of the median body length of C. megacephala and C. rufifacies larvae in different seasons of the years 2000–2001 in Thailand revealed that both species developed rapidly in the summer; pupariation of C. rufifacies initiated at 144 h, while C. megacephala initiated pupariation at 156 h. This information is potentially useful for estimating the postmortem interval of a corpse in forensic investigations, where the corpse becomes infesting with these fly species. This work has been presented at the 21st Pacific Science Congress held in Okinawa, Japan, in 12–18 June, 2007.     

<Emphasis Type="Italic">Sarcocystis</Emphasis> sp. from the herring gull (<Emphasis Type="Italic">Larus argentatus</Emphasis>) identity to <Emphasis Type="Italic">Sarcocystis wobeseri</Emphasis> based on cyst morphology and DNA results

Having studied 11 herring gulls (Larus argentatus) Sarcocystis cysts were found in neck and leg muscles of 4 birds. One type of sarcocysts (cyst type I) that have a thin (∼1.0 μm), smooth, or slightly wavy cyst wall without clearly visible protrusions and small (6.0–8.0 μm) lancet- or banana-shaped cystozoites was identified by the light microscopy. Sarcocysts extracted from one herring gull were used for electron microscopy and DNA analysis. Ultrastructurally, Sarcocystis sp. from the herring gull had the same tissue cyst wall type-1 as S. calchasi, S. columbae, and S. wobeseri parasitizing in birds. According to first internal transcribed spacer (ITS-1) region, 18S rRNA and 28S rRNA gene sequences, Sarcocystis sp. from the herring gull belongs to S. wobeseri. Nevertheless, without evidences of cross-transmission experiment sarcocysts extracted from herring gull at present time are named as S. wobeseri-like.     

<Emphasis Type="Italic">Dracunculus brasiliensis</Emphasis> sp. n. (Nematoda: Dracunculidae) from the anaconda, <Emphasis Type="Italic">Eunectes murinus</Emphasis> (Ophidia: Boidae)

Dracunculus brasiliensis sp. n. (Dracunculidae), is described based on a single female specimen found in the body cavity of the anaconda, Eunectes murinus (L.) (Ophidia: Boidae), from the Mexiana Island, Amazon River delta, Brazil and one female previously recorded from the subcutaneous tissue of this host species imported from South America into Europe (ZOO in the Czech Republic). The new species is characterised mainly by markedly large, anteriorly protruding dorsal and ventral double papillae of the internal circle and small lateral papillae of the same circle, a widely rounded caudal end, the excretory pore situated just posterior to the nerve ring, a distinctly transversely striated cuticle and by the length (396–429 μm) of larvae from uterus. This is the first species of Dracunculus described from reptiles in South America.     

Occurrence of cystacanths of <Emphasis Type="Italic">Plagiorhynchus cylindraceus</Emphasis> (Acanthocephala) in the terrestrial isopods <Emphasis Type="Italic">Trachelipus squamuliger</Emphasis> and <Emphasis Type="Italic">Armadillidium vulgare</Emphasis> (Oniscidea) in Bulgaria

In total, 2097 individuals of Trachelipus squamuliger and 20 individuals of Armadillidium vulgare from four habitats (three woodland sites and one pasture) in the region of Stara Zagora, Bulgaria, were examined for the presence of cystacanths of Plagiorhynchus cylindraceus, a common acanthocephalan parasite of passerine birds. In T. squamuliger from woodland habitats, cystacanths were found with prevalence 4.0–9.3%, intensity 1–5 (mean 1.22–1.57) and mean abundance 0.057–0.113. No significant differences were observed between infections in males and females of T. squamuliger. None of the T. squamuliger individuals from the pasture examined was infected. Out of 48 infected females of T. squamuliger, only one had developed eggs (in agreement with previous studies revealing the negative effect of the cystacanths on the development of female gonads of woodlice). One individual of A. vulgare was infected with a single cystacanth. The occurrence of P. cylindraceus in T. squamuliger is a new host record.     

Larvicidal activity of silver nanoparticles synthesized using <Emphasis Type="Italic">Plumeria rubra</Emphasis> plant latex against <Emphasis Type="Italic">Aedes aegypti</Emphasis> and <Emphasis Type="Italic">Anopheles stephensi</Emphasis>

In the present study activity of silver nanoparticles (AgNPs) synthesized using Plumeria rubra plant latex against second and fourth larval instar of Aedes aegypti and Anopheles stephensi was determined. Range of concentrations of synthesized AgNps (10, 5, 2.5, 1.25, 0.625, 0.3125 ppm) and aqueous crude latex (1,000, 500, 250, 125, 62.50, 31.25 ppm) were tested against larvae of A. aegypti and A. Stephensi. The synthesized AgNps from P. rubra latex were highly toxic than crude latex extract in both mosquito species. The LC₅₀ values for second and fourth larval instars after 24 h of crude latex exposure were 1.49, 1.82 ppm against A. aegypti and 1.10, 1.74 ppm against A. stephensi respectively. These figures were 181.67, 287.49 ppm against A. aegypti and 143.69, 170.58 ppm against A. stephensi respectively for crude latex extract. The mortality rates were positively correlated with the concentration of AgNPs. The characterization studies of synthesized AgNPs by UV–Vis spectrophotometry, transmission electron microscopy (TEM), Particle size analysis (PSA) and zeta potential confirmed the spherical shape and size (32–200 nm) of silver nanoparticles alongwith stability. Toxicity studies carried out against non-target fish species Poecilia reticulata, the most common organism in the habitats of A. aegypti and A. stephensi showed no toxicity at LC₅₀ and LC₉₀ doses of the AgNPs. This is the first report on mosquito larvicidal activity of latex synthesized nanoparticles.     

<Emphasis Type="Italic">Sarcocystis</Emphasis> sp. from the goldeneye (<Emphasis Type="Italic">Bucephala clangula</Emphasis>) and the mallard (<Emphasis Type="Italic">Anas platyrhynchos</Emphasis>): cyst morphology and ribosomal DNA analysis

By light microscopy, cysts of Sarcocystis sp. (cyst type I) from the goldeneye (Bucephala clangula) seemed filamentous with a smooth and thin (<1 μm) cyst wall. Ultrastructurally, the cyst wall surface was irregular with minute undulations of the primary cyst wall. These sarcocysts had type-1 cyst wall. Cystozoites were banana-shaped and measured 7.0–8.5 μm in length. By light microscopy, cysts of Sarcocystis sp. (cyst type II) from the mallard (Anas platyrhynchos) were ribbon-shaped, very long, and thin. On the surface of the wall (up to 1.5 μm), they had palisade-like villar protrusions closely crowded together. Electron micrographs showed villar protrusions (up to 1.3 μm in length) different in size and shape. The latter had short microprojections especially obvious in the oblique sections. Cystozoites were slightly bent with blunt ends, broader at one end, and measured 13.0–16.1 × 1.8–2.5 μm. Phylogenetic analysis based on the comparison of partial 28S rRNR gene sequences of Sarcocystis sp. (cyst type II) derived from the mallard, Sarcocystis sp. (cyst type I) and Sarcocystis sp. (cyst type III) both derived from the white-fronted goose (Anser albifrons) suggested that these sequences belonged to separate Sarcocystis species.     

The comparative morphology of <Emphasis Type="Italic">Marshallagia marshalli</Emphasis> and <Emphasis Type="Italic">Ostertagia occidentali</Emphasis>s (Nematoda: Strongylida,Trichostrongylidae) by scanning electron microscopy

The systematics of the Ostertagiinae is unsettled with no agreement on how many genera and species are present in cattle and sheep. Ten species of Ostertagiinae are commonly parasitic in cattle and sheep. In the global fauna, six of 13 ostertagiine genera are endemic to Iran. The life cycle of Ostertaginae is direct and ingested third-stage larvae after exsheatment in the rumen, penetrate the gastric glands in the abomasal mucosa where two parasitic moults occur before the L5 emerges from the gland. In the present work, Marshallagia marshalli and Ostertagia occidentalis, collected from the abomasums of sheep from Mashhad, Iran, is described. The association of light and scanning electron microscopy (SEM) allowed a detailed analysis of the morphology and ultrastructure of these nematodes. The male body length of M. marshalli and O. occidentalis were 9.3–10.20 and 9.60–10.50 mm, respectively. The female body length of M. marshalli and O. occidentalis were 10.10–15.30 and 10.4–15.70 mm, respectively. One of cervical papillae is seen 333 and 250 μm from the anterior end of male and female body surface in O. occidentalis and 287.5 and 200 μm from the anterior end of male and female body surface in M. marshalli, respectively. The size of cervical papillae is 13.3 μm in male and 10 μm in female in O. occidentalis and 9.33 μm in male and 8.57 μm in female in M. marshalli. Some other taxonomic features of M. marshalli and O. occidentalis, such as details of cephalic region, the system of longitudinal and surface cuticular ridges (synlophe), the orientation of rays of the copulatory bursa, localization of vulva, morphology of vulvar flap, and posterior end of females are also documented by SEM.     

A new species of <Emphasis Type="Italic">Falcaustra</Emphasis> (Nematoda,Kathlaniidae) from <Emphasis Type="Italic">Cnemaspis</Emphasis> aff. <Emphasis Type="Italic">tro pidogaster</Emphasis> (Squamata,Gekkonidae) from Sri Lanka

Falcaustra desilvai sp. nov. (Ascaridida, Kathlaniidae) from the large intestine of Cnemaspis aff. tropidogaster (Squamata, Gekkonidae) is described and illustrated. Falcaustra desilvai represents the 4th nematode species from Sri Lanka to be assigned to the genus and is distinguished from other Sri Lankan species by the distribution pattern of caudal papillae (12 precloacal, 2 adcloacal, 10 postcloacal, and 1 median), length of spicules (956–1046 μm) and absence of a pseudosucker.     

Characterization of <Emphasis Type="Italic">Taenia solium</Emphasis> cysticerci microsomal glutathione <Emphasis Type="Italic">S</Emphasis>-transferase activity

Glutathione S-transferase activity has been shown to be associated with the microsomal fraction of Taenia solium. Electron microscopy and subcellular enzyme markers indicate the purity of the microsomal fraction that contains the glutathione S-transferase activity. T. solium microsomes were solubilized under conditions used to solubilize integral microsomal proteins. This procedure proved necessary to obtain enzymatic activity. To characterize this parasite enzyme activity, several substrates and inhibitors were used. The optimum activity for microsomal glutathione S-transferase was found to be pH 6.6, with a specific enzyme activity of 0.9, 0.1, 0.067, 0.03, and 0.05 μmol min⁻¹ mg⁻¹ with the substrates 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene, 4-hydroxynonenal, 2,4-hexadienal, and trans-2-nonenal, respectively. No activity of glutathione peroxidase was observed. T. solium microsomes had an _app K _{m (GSH)} = 0.161 μM, _app K _{m (CDNB)} = 14.5 μM, and _app V _max of 0.15 and 27.9 μmol min⁻¹ mg⁻¹ for GSH and CDNB, respectively. T. solium microsomes were inhibited by several glutathione S-transferase enzyme inhibitors, and it was possible to establish a simple inhibition system as well as corresponding K _i ’s for each inhibitor. These results indicate that the T. solium microsomal glutathione S-transferase is different from the parasite cytoplasmic enzymes that catalyze similar reactions.     

The life cycle of <Emphasis Type="Italic">Stephanoprora aylacostoma</Emphasis> n.sp. (Digenea: Echinostomatidae), parasite of the threatened snail <Emphasis Type="Italic">Aylacostoma chloroticum</Emphasis> (Prosobranchia,Thiaridae), in Argentina

A new species, Stephanoprora aylacostoma is described and its life cycle was resolved experimentally. The prosobranch snail Aylacostoma chloroticum Hylton Scott, collected in the Yacyretá dam, Province of Misiones, Argentina, was found naturally infected with large-tailed cercariae possessing a prepharyngeal body and corpuscles in the excretory system but lacking collar spines. Metacercariae, which encysted on the gills of experimentally infected fishes Cnesterodon decemmaculatus (Jenyns) and Poecilia reticulata (Peters) (Poecilidae), developed collar spines after 10 days. Tetragonopterid fishes Moenckhausia dichroura (Kner), Astyanax erythropterus (Holmberg) and Hyphesobrycon serpae (Durbin in Eigenmann) were found infected naturally. Sexually mature adults were recovered from domestic chicks at day 7 post-exposure. Eggs shed in chick faeces developed to miracidia within 13–15 days; sporocysts were found on the gills of snails. The new species differs from other species of the genus in its larger eggs, in the smaller, slender body and smaller collar spines of the adult and in the morphological and biological features of the larval stages.     

Efficacy of female <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis> with entomopathogenic fungus <Emphasis Type="Italic">Fusarium pallidoroseum</Emphasis>

This study was conducted to isolate and identify natural entomopathogenic fungi from female Culex quinquefasciatus and to test their adulticidal activity. Field-collected female C. quinquefasciatus died early and were placed on a Saboraud’s dextrose agar plates for growth and isolation of natural entomopathogenic fungi. The plates were maintained in an incubator at 24 ± 2°C for 3 days. Four fungal species were isolated in two genera namely, Aspergillus and Fusarium. The identified fungal species were A. niger, A. flavus, A. nidulans var acristatus (ITCC-6327.04), and F. pallidoroseum (ITCC-6324.06). Adult bioassays were carried out using spore-impregnated paper in WHO-holding tubes. F. pallidoroseum was found to be more effective than the others. Exposure of C. quinquefasciatus to spores of A. flavus and A. niger for 4 h caused 5.53% and 5.51% mortality in the mosquitoes within a week, respectively. All the female C. quinquefasciatus were killed within 4 days of exposure to F. pallidoroseum at a concentration of 1.11 × 10¹⁰ conidia per m². Significant difference of longevity was observed between the F. pallidoroseum-treated C. quinquefasciatus and control mosquitoes. The LT₅₀ of F. pallidoroseum was 2.08 days for 4 h exposure to C. quinquefasciatus. Results of the present study confirm that F. pallidoroseum is one of the alternative biological control agents of adult mosquitoes.     

<Emphasis Type="Italic">Eimeria biarmicus</Emphasis> sp.n. (Apicomplexa: Eimeriidae) infecting falcons from the genus <Emphasis Type="Italic">Falco</Emphasis> in Saudi Arabia

The oocysts of Eimeria biarmicus sp. n. were described from the feces of the lanner falcon, Falco biarmicus, collected from the falcon market in Riyadh City, Saudi Arabia. The prevalence of infection was 5% (2/40). The majority of the oocysts examined had completed sporulation within 84 h at 24 ± 2°C. Sporulated oocysts are ovoid in shape, measuring 22.4 × 17.9 (20.5–24.7 × 15.8–18.5) μm; shape index (L/W) is 1.25 (1.14–1.36) μm. The oocyst wall is smooth and bi-layered. Micropyle and oocyst residuum are absent. A polar granule is present, consisting of 2–4 globules. Sporocysts are ovoid, 10.1 × 6.1 (9.4–11.2 × 5.4–6.8) μm; with a smooth single-layered wall and a minute Stieda body, but there is no substieda body. The sporocyst residuum consists of numerous small granules. Sporozoites are comma shaped, each contains two refractile bodies. E. biarmicus sp. n. is the second eimerian species described from F. biarmicus.     

<Emphasis Type="Italic">Mexiconema africanum</Emphasis> sp. n. (Nematoda: Daniconematidae) from the catfish <Emphasis Type="Italic">Auchenoglanis occidentalis</Emphasis> from Lake Turkana,Kenya

A new species of dracunculoid nematode, Mexiconema africanum sp. n. (Daniconematidae), is described from the abdominal cavity and the intestine (rarely also the gall bladder) of the catfish Auchenoglanis occidentalis (Valenciennes) (Claroteidae, Siluriformes) from Lake Turkana, Kenya. The new species differs from two other congeners mainly in the absence of two large cell nuclei in the glandular oesophagus, presence of well-developed lateral cephalic elevations, more numerous (14) cephalic papillae and a much longer body of the gravid female (18–22 mm); from Mexiconema cichlasomae Moravec, Vidal and Salgado Maldonado, 1992 also in less numerous (two) caudal processes and a different arrangement of genital papillae in the male. M. africanum is the first representative of the dracunculoid family Daniconematidae described from Africa.     

A novel nuclear marker,Pm-int9, for phylogenetic studies of <Emphasis Type="Italic">Opisthorchis felineus</Emphasis>, <Emphasis Type="Italic">Opisthorchis viverrini</Emphasis>, and <Emphasis Type="Italic">Clonorchis sinensis</Emphasis> (Opisthorchiidae,Trematoda)

Opisthorchis felineus, O. viverrini, and Clonorchis sinensis, the trematodes of the family Opisthorchiidae, are important human parasites. Two previous studies (Kang et al. Parasitol Int 57:191–197, 2008; Katokhin et al. Dokl Biochem Biophys 421:214–217, 2008) have provided evidence using ribosomal and mitochondrial sequences that O. viverrini, O. felineus, and C. sinensis are closely related. We developed a novel nuclear marker, Pm-int9, which included the ninth intron of the paramyosin gene and flanking exon sequences. Samples of O. felineus from four localities of West Siberia, C. sinensis from the Russian Far East, and O. viverrini from Thailand were genotyped by Pm-int9. Little variation was detected in exon sequences, however, intron sequences turned out to be more variable than ribosomal internal transcribed spacers. We can conclude that Pm-int9 is valuable for interspecific variation studies. Phylogenetic analysis based on Pm-int9 revealed that O. viverrini and C. sinensis were closer to each other than either of them to O. felineus, supporting the opinion that C. sinensis should be considered the sister species of Opisthorchis spp.     

<Emphasis Type="Italic">Falcaustra sinensis</Emphasis> sp. nov. (Nematoda,Kathlaniidae) from <Emphasis Type="Italic">Indotestudo elongata</Emphasis> in China

Falcaustra sinensis sp. nov. collected from the feces of Indotestudo elongata (Blyth) is described and illustrated. The new species can be distinguished from the congeners by the arrangement of caudal papillae of the male (6 pairs precloacal, 4 pairs postcloacal, 1 median), length of spicules (3.84–4.74 mm) and the presence of pseudosucker.     

Characterisation of an isolate of <Emphasis Type="Italic">Narcissus degeneration virus</Emphasis> from Chinese narcissus (<Emphasis Type="Italic">Narcissus tazetta</Emphasis> var. <Emphasis Type="Italic">chinensis</Emphasis>)

Summary. A potyvirus from Chinese narcissus was transmitted mechanically to three species of Narcissus and to Lycoris radiata but not to 22 other test species. In western blot, the coat protein reacted strongly with Narcissus degeneration virus (UK isolate) antiserum. Antiserum raised to the Chinese virus did not react with eighteen other potyviruses. The complete nucleotide sequence (9816 nt) had the typical genome organisation for a member of the genus Potyvirus. Sequence comparisons and phylogenetic analysis showed that the Chinese virus was different from all previously sequenced potyviruses but distantly related to onion yellow dwarf and shallot yellow stripe viruses.     

A new species of <Emphasis Type="Italic">Hysterothylacium</Emphasis> (Ascaridoidea,Anisakidae) parasitic in <Emphasis Type="Italic">Zenopsis conchifer</Emphasis> (Zeiformes,Zeidae) from Argentinean waters

A new species of parasitic nematode, Hysterothylacium spirale sp. nov. (Ascaridoidea, Anisakidae), is described based on specimens collected from the intestine and pyloric caeca of the silvery john dory Zenopsis conchifer (Lowe) (Zeiformes, Zeidae), from the Argentinean Shelf (35°05′–40°46′S, 53°03′–58°07′W). Among the 66 valid species described so far in the genus, the new species most closely resembles H. zenis (Baylis, 1929). Both species, apparently specific for fishes of the family Zeidae, share the shape of the dorsal lip, the long expanded lateral alae originating from subventral interlabia and the ornamentation of the tail tip, as well as general morphometry. The combination of these shared features distinguishes both species from all congeners so far known. However, the new species is distinguishable from H. zenis by having shorter interlabia, and consequently the lateral alae originating more posteriorly, shorter spicules, a smaller number of postcloacal papillae and the presence of two pairs of double postcloacal papillae.     

<Emphasis Type="Italic">Kudoa unicapsula</Emphasis> n. sp. (Myxosporea: Kudoidae) a parasite of the Mediterranean mullets <Emphasis Type="Italic">Liza ramada</Emphasis> and <Emphasis Type="Italic">L. aurata</Emphasis> (Teleostei: Mugilidae)

A new multivalvulid myxozoan parasite, Kudoa unicapsula n. sp., is described from the intestinal mesentery, intestine and pyloric caeca of the thin-lipped grey mullet Liza ramada (Risso 1826) and the golden grey mullet L. aurata (Risso, 1810) from the Mediterranean coastal waters of Spain. It is characterized by the presence of elongated, rice corn-like white cysts of 0.47–0.56 × 0.18–0.38 mm, filled with tetracapsulate, slightly asymmetric spores, rectangular in apical view and tear-shaped in lateral view with four polar capsules of considerably different size and slightly unequal spore valves with rounded edges, overlapping each other on the apex of the spore. One large polar capsule includes a polar filament coiled in two to three turns, and the other three polar capsules, which are very small, posses only a rudimental filament. Both light and electron microscopy data showed that this species differs from all previously described Kudoa spp. with unequal polar capsules. The molecular analysis based on 18S and 28S ribosomal deoxyribonucleic acid DNA sequence data of K. unicapsula n. sp. indicates a close relationship and thus phylogenetic clustering together with K. trifolia, a myxozoan from the same host and the same geographical location.     

<Emphasis Type="Italic">Myxobolus honghuensis</Emphasis> n. sp. (Myxosporea: Bivalvulida) parasitizing the pharynx of allogynogenetic gibel carp <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis> (Bloch) from Honghu Lake,China

Myxobolus honghuensis n. sp. is described from allogynogenetic gibel carp Carassius auratus gibelio (Bloch), during a survey of myxosporean parasites in Honghu Lake, Hubei Province, China. It is characterized by the presence of round plasmodia of 5–12 mm in diameter in the pharynx of host. Mature spores of M. honghuensis were pyriform in frontal view and anterior pointed with bluntly round posterior, they measured 16.9 ± 0.5 (15.1–19.5) μm long, 10.4 ± 0.4 (9.0–11.3) μm wide, and 8.4 ± 0.4 (7.9–9.1) μm thick. Two polar capsules were pyriform and slightly unequal with larger polar capsule 8.4 ± 0.4 (7.6–10.2) μm × 3.9 ± 0.2 (3.0–4.5) μm and smaller capsule 7.9 ± 0.2 (7.0–9.3) μm × 3.7 ± 0.3 (2.8–4.1) μm. Polar filaments coiled with seven to eight turns. Both morphology and DNA sequence data revealed that M. honghuensis n. sp. was distinct from other described Myxobolus species. Phylogenetic analysis placed M. honghuensis n. sp. in a clade of gill-infecting myxobolids.