Morphological and molecular characterization of Ceratomyxa binhthuanensis n. sp. (Myxosporea: Ceratomyxidae) from the gall bladder of blacktip grouper Epinephelus fasciatus (Perciformes: Serranidae) in the East Sea of Vietnam.

A new myxozoan species, Ceratomyxa binhthuanensis n. sp. (Myxosporea: Ceratomyxidae), was found in the gall bladder of blacktip grouper Epinephelus fasciatus (Perciformes: Serranidae) in the East Sea of Vietnam. Myxospores were observed floating free in the gall bladder of 3 out of 20 fish examined (15%). Mature myxospores were elongate and slightly crescent-shaped and measured 12.2 ± 1.3 (10.8-16.0) µm in thickness and 5.8 ± 0.6 (4.8-6.9) µm in length, with two smooth equal shell valves. The two polar capsules were spherical and equal in size, measuring 2.6 ± 0.3 (2.3-2.9) µm in diameter. The posterior angle was slightly concave, 153.7° ± 5.6° (148.9°-166.0°). Molecular analysis of SSU rDNA sequence showed that Ceratomyxa binhthuanensis n. sp. differs from other Ceratomyxa spp. available in GenBank. Phylogenetic analysis indicated that C. binhthuanensis n. sp. was closely related to three species, Ceratomyxa nolani, Ceratomyxa yokoyamai, and Ceratomyxa cutmorei, which also infect fish hosts of the genus Epinephelus.

Description and Molecular Differentiation of a New Skrjabinoptera (Nematode: Physalopteridae) from Eutropis macularia (Sauria: Scincidae) in North-Central Vietnam.

Skrjabinoptera vietnamensis n. sp. is described from specimens recovered from the stomach of Eutropis macularia in north-central Vietnam. The new species is characterized by the medium-sized male worms (6.7-8.7 mm in length and 154-182 µm in width) relative to known members of the genus, 2 pointed spicules of unequal length (87-112 µm and 56-72 µm in length), and 10 pairs of caudal papillae. Female worms are larger than male worms (10.7-18.4 mm in length and 264-411 µm in width), with the vulva situated in the anterior part, and embryonated, elliptical eggs, 35-46 µm long by 20-24 µm wide. Skrjabinoptera vietnamensis n. sp. represents the ninth species assigned to the genus and the first species recorded from the Oriental region. Partial sequences of the 18S ribosomal RNA gene (rDNA), and cytochrome c oxidase subunit 1 (COI) are provided for the new species. The molecular phylogenetic position of the genus Skrjabinoptera is briefly discussed.

Description and Molecular Differentiation of a New Falcaustra (Nematode: Kathlaniidae) from the Indochinese Water Dragon, Physignathus cocincinus (Squamata: Agamidae) in North-central Vietnam.

Falcaustra vietnamensis n. sp. is described from the small intestine of Physignathus cocincinus from north-central Vietnam. The new species is characterized by the large male worms (20.2-28.8 mm in length and 557-724 µm in width) relative to known members of the genus, 2 sharply pointed alate spicules of equal length (1,128-1,256 µm in length), gubernaculum including 2 separate pieces, 1 ventral with a pointed distal end and 1 dorsal with a blunt distal end (164-192 µm and 155-172 µm in length, respectively), and 12 pairs of caudal papillae. Female worms are larger than male worms (24.2-34.1 mm in length and 532-735 µm in width), with the vulva situated in the posterior half of body, and elliptical eggs, 60-70 µm long by 42-47 µm wide. Falcaustra vietnamensis n. sp. represents the 38th species assigned to the genus and the third species recorded from a lizard host in the Oriental biogeographical region. Partial sequences of the 18S ribosomal RNA gene (rDNA), internal transcribed spacer regions (ITS), and cytochrome c oxidase subunit 1 (COI) are provided for the new species. The molecular phylogenetic position of the genus Falcaustra is briefly discussed.

Morphological and Molecular Description of Rhadinorhynchus hiansi Soota and Bhattacharya, 1981 (Acanthocephala: Rhadinorhynchidae) from Marine Fish off the Pacific Coast of Vietnam.

Rhadinorhynchus hiansi Soota and Bhattacharya, 1981 , has remained unknown since its original incomplete description from 2 male specimens collected from the flat needlefish Ablennes hians Valenciennes (Belonidae) off Trivandrum, Kerala, India. Recent collections of fishes along the Pacific coast of Vietnam in 2016 and 2017 produced many specimens of the same species from the striped bonito Sarda orientalis Temminck and Schlegel (Scombridae) off the southern Pacific coast of Vietnam at Nha Trang. We describe females for the first time, assign a female allotype status, and provide an expanded description of males from a larger collection completing missing information on hooks and hook roots, receptacle, lemnisci, cement glands, Saefftigen's pouch, and trunk spines. Specimens of R. hiansi characteristically have no dorsal spines in the posterior field of trunk spines and a long proboscis with 36-48 dorso-ventrally differentiated proboscis hooks per row becoming progressively smaller posteriorly then increasing in size near the posterior end to a maximum at the posterior-most ring. Trunk, testes, and lemnisci in our specimens were considerably larger than those reported in the original description, but the proboscis was relatively smaller. The females had long reproductive system and corrugated elliptic eggs without polar prolongation of fertilization membrane. Energy Dispersive X-ray Analysis (EDXA) demonstrates high levels of calcium and phosphorus in large gallium cut hooks and high levels of sulfur in tip cuts of large and small hooks and in spines. This EDXA pattern is a characteristic fingerprint of R. hiansi. The molecular profile of R. hiansi is described from 18S rDNA and COI genes, and phylogenetic relationships with most closely related species are discussed.

Descriptions of two new acanthocephalans (Rhadinorhynchidae) from marine fish off the Pacific coast of Vietnam.

Two rhadinorhynchid acanthocephalans are described from marine fishes off the Pacific coast of Vietnam. Sclerocollum neorubrimaris n. sp. (Gorgorhynchinae Van Cleave & Lincicome, 1940) is described from the spine-foot rabbitfish Siganus guttatus (Bloch) (Siganidae) off Nha Trang. The new species is similar to the type-species, Sclerocollum rubrimaris Schmidt & Paperna, 1978 in most measurements and counts but has a posterior cephalic ganglion, a sub-ventral female gonopore, and one dorsal paravaginal filament bundle. In S. rubrimaris, the cephalic ganglion is near the middle of the receptacle, the female gonopore is terminal with two paravaginal filament bundles. The new species is distinguished from three other species of Sclerocollum Schmidt & Paperna, 1978 in proboscis hook formulas and the position of the female gonopore and cephalic ganglion. The issue of the importance of the position of the cephalic ganglion at the generic and sub-generic levels in the Acanthocephala is discussed. Females of Australorhynchus multispinosus n. sp. (Gorgorhynchinae) are described from the red cornetfish Fistularia petimba Lacépède off Nha Trang in the Pacific south. It is distinguished from the only other species of the genus, Australorhynchus tetramorphacanthus Lebedev, 1967, by having more trunk spines extending beyond the level of the proboscis receptacle and a smaller proboscis with considerably fewer hooks.

Descriptions of acanthocephalans, Cathayacanthus spinitruncatus (Rhadinorhynchidae) male and Pararhadinorhynchus magnus n. sp. (Diplosentidae), from marine fish of Vietnam, with notes on Heterosentis holospinus (Arhythmacanthidae).

Males of Cathayacanthus spinitruncatus Amin, Heckmann & Ha, 2014 (Rhadinorhynchinae Lühe, 1912) are described for the first time from Leiognathus equulus in Hai Phong and Nha Trang and from pony fish Nuchequula flavaxilla in Quang Ninh in the Pacific waters of Vietnam. The male allotype status is designated. Males of C. spinitruncatus are smaller and have fewer and smaller proboscis hooks and trunk spines than females. The male reproductive structures are in the posterior fifth of the trunk and with 6 club-shaped cement glands gradually merging into 6 independent cement gland ducts. The proboscis receptacle is more than half as long as the trunk and with a cephalic ganglion at its anterior end. In females, the receptacle is only about one fifth the length of the trunk. Specimens described as Cathayacanthus bagarii Moravec & Sey, 1989 were shown to have been wrongly assigned to Cathayacanthus. Pararhadinorhynchus magnus n. sp. (Diplosentidae) is described from Scatophagus argus off Hai Phong in the Gulf of Tonkin. It is the third species of the genus and is readily distinguished from the Australian species by having a considerably larger trunk and male reproductive structures, and more proboscis hooks. X-ray microanalysis (EDAX) of intact and gallium-cut hooks of P. magnus showed high calcium and phosphate mainly in the central core. Specimens of Heterosentis holospinus Amin, Heckmann & Ha, 2011 (Arhythmacanthidae) are also reported from L. equulus off Quang Binh, new host and locality records.

Descriptions of Neorhadinorhynchus nudum (Cavisomidae) and Heterosentis paraholospinus N. SP. (Arhythmacanthidae) (Acanthocephala) From Fish Along the Pacific Coast of Vietnam, With Notes On Biogeography.

Two species of acanthocephalans are described from fishes caught along the Pacific coast off eastern Vietnam in 2016: (1) Neorhadinorhynchus nudum ( Harada, 1938 ) Yamaguti, 1939 (Cavisomidae) from the frigate tuna Auxis thazard (Lacépède) (Scombridae) in Nha Trang, Pacific south Vietnam, and (2) Heterosentis paraholospinus n. sp. (Arhythmacanthidae) from 3 species of fish: the common ponyfish Leiognathus equulus (Forsskål) (Leiognathidae) and the torpedo scad Megalaspis cordyla (Linn.) (Carangidae) off Nha Trang and Binh Thuân, respectively, and the yellowspotted ponyfish Nuchequula flavaxilla Kimura, Kimura, and Ikejima (Leiognathidae) at Quang Ninh in Pacific waters of Vietnam. Females of N. nudum are fully described for the first time in the Pacific Ocean. Heterosentis paraholospinus n. sp. is similar to Heterosentis holospinus Amin, Heckmann, and Ha, 2011, in having an unspined anterior trunk cone, nucleated pouch at the posterior end of the receptacle, and spines covering the entire trunk, but it differs in having a long cylindrical trunk, lemnisci much longer than the receptacle, more and different-sized proboscis hooks, and subterminal female gonopore. Biogeographical notes are discussed for both genera.

Application of quantitative real-time PCR compared to filtration methods for the enumeration of Escherichia coli in surface waters within Vietnam.

Surface water samples in Vietnam were collected from the Saigon River, rural and suburban canals, and urban runoff canals in Ho Chi Minh City, Vietnam, and were processed to enumerate Escherichia coli. Quantification was done through membrane filtration and quantitative real-time polymerase chain reaction (PCR). Mean log colony-forming unit (CFU)/100 ml E. coli counts in the dry season for river/suburban canals and urban canals were log 2.8 and 3.7, respectively, using a membrane filtration method, while using Taqman quantitative real-time PCR they were log 2.4 and 2.8 for river/suburban canals and urban canals, respectively. For the wet season, data determined by the membrane filtration method in river/suburban canals and urban canals samples had mean counts of log 3.7 and 4.1, respectively. While mean log CFU/100 ml counts in the wet season using quantitative PCR were log 3 and 2, respectively. Additionally, the urban canal samples were significantly lower than those determined by conventional culture methods for the wet season. These results show that while quantitative real-time PCR can be used to determine levels of fecal indicator bacteria in surface waters, there are some limitations to its application and it may be impacted by sources of runoff based on surveyed samples.

Real-time PCR detection and phylogenetic relationships of Neorickettsia spp. in digeneans from Egypt, Philippines, Thailand, Vietnam and the United States.

Neorickettsia (Rickettsiales, Anaplasmataceae) is a genus of obligate intracellular bacterial endosymbionts of digeneans (Platyhelminthes, Digenea). Some Neorickettsia are able to invade cells of the digenean's vertebrate host and are known to cause diseases of domestic animals, wildlife, and humans. In this study we report the results of screening digenean samples for Neorickettsia collected from bats in Egypt and Mindoro Island, Philippines, snails and fishes from Thailand, and fishes from Vietnam and the USA. Neorickettsia were detected using a real-time PCR protocol targeting a 152bp fragment of the heat shock protein coding gene, GroEL, and verified with nested PCR and sequencing of a 1853bp long region of the GroESL operon and a 1371bp long region of 16S rRNA. Eight unique genotypes of Neorickettsia were obtained from digenean samples. Neorickettsia sp. 8 obtained from Lecithodendrium sp. from Egypt; Neorickettsia sp. 9 and 10 obtained from two species of Paralecithodendrium from Mindoro, Philippines; Neorickettsia sp. 11 from Lecithodendrium sp. and Neorickettsia sp. 4 (previously identified from Saccocoelioides lizae, from China) from Thailand; Neorickettsia sp. 12 from Dicrogaster sp. Florida, USA; Neorickettsia sp. 13 and SF agent from Vietnam. Sequence comparison and phylogenetic analysis demonstrated that the forms, provisionally named Neorickettsia sp. 8-13, represent new genotypes. We have for the first time detected Neorickettsia in a digenean from Egypt (and the African continent as a whole), the Philippines, Thailand and Vietnam based on PCR and sequencing evidence. Our findings suggest that further surveys from the African continent, SE Asia, and island countries are likely to reveal new Neorickettsia lineages as well as new digenean host associations.

Current status of fish-borne zoonotic trematode infections in Gia Vien district, Ninh Binh province, Vietnam.

BACKGROUND: Ninh Binh province is known as an endemic area of fish-borne zoonotic trematode (FZT) transmission in Vietnam. A cross-sectional study was conducted in Gia Minh and Gia Thinh communes of Gia Vien district, Ninh Binh province to investigate the infections with different stages of FZT in various host species. METHODS: Faecal samples from 1,857 humans were examined for trematode eggs using the Kato-Katz method, while faecal samples from 104 dogs, 100 cats, and 100 pigs were examined using the Formalin-ethyl acetate concentration technique (FECT). A total of 483 specimens of freshwater fish, representing 9 species, were examined for metacercariae using the artificial digestion method. Three methods of cercarial detection (shedding, crushing and cutting) were applied for examination of 3,972 specimens of freshwater snails, representing 7 species. All relevant data e.g. location, sex, age group, animal species, and habitat were recorded for statistical analyses. RESULTS: Helminth eggs were found in 65.5% of the human faecal samples, including 20.5% of faecal samples containing small trematode eggs. Infection with small trematodes differed among communes, age groups and sexes. Eggs of small trematodes were found in 32.7% of faecal samples from dogs, 49.0% from cats and 13.0% from pigs. The difference in prevalences and intensities were significant among species of animals but did not differ between the two communes. All fish species were infected with FZT, with an average prevalence of 56.1% and a mean intensity of 33.245 metacercariae per gram. Prevalence and intensity in fish differed significantly among cummunes and fish groups. Six species of zoonotic trematodes were identified. Metacercariae of the small liver fluke, Clonorchis sinensis, was only found in Hemiculter leucisculus. A total of 9 specimens from two snail species, Melanoides tuberculata and Bithynia fuchsiana, were infected with trematodes and four cercarial types were detected in the study sites. CONCLUSIONS: We conclude that Gia Minh and Gia Thinh communes are continuing to be hot-spot endemic areas of FZT and other helminths infections where the habit of eating raw fish by the local people is still present.

Acanthocephalans from fishes and amphibians in Vietnam, with descriptions of five new species.

Eight species of acanthocephalans are reported, and five are new. Specimens of Neoechinorhynchus (Hebesoma) manubrianus Amin, Ha & Ha, 2011 were similar to the original description. Neoechinorhynchus (Hebesoma) spiramuscularis n. sp. (Neoechinorhynchidae), from Xenocypris davidi, has a unique proboscis receptacle wrapped in a spiral muscular layer, and an undulating flask-shaped lemnisci, as well as double para-receptacle structures. Heterosentis mongcai n. sp. (Arhythmacanthidae), from Acreichthys sp., has a small fusiform trunk with an unarmed cone and anterior trunk spines, and a proboscis with two circles of rooted apical hooks and 3-4 circles of rooted posterior spines as well as a para-receptacle-like structure at the posterior end. The poorly known Filisoma indicum Van Cleave, 1928 is fully described and illustrated for the first time. Acanthocephalus parallelcementglandatus n. sp. (Echinorhynchidae), from Clarias batrachus, is distinguished from other species of Acanthocephalus by its small fusiform trunk and parallel tubular cement glands. Pseudoacanthocephalus coniformis n. sp. (Echinorhynchidae), from Hylarana sp., is distinguished from other species by having an anterior trunk collar and staggered prominent filiform cement glands, among other features. Cathayacanthus spinitruncatus n. sp. (Rhadinorhynchidae), from Leiognathus equulus, is distinguished from the only two known species of the genus by having a very long and slender proboscis with more than 50 hooks per row and a totally spined trunk. The generic diagnosis of Cathayacanthus Golvan, 1969 is emended. Rhadinorhynchus johnstoni Golvan, 1969 (Rhadinorhynchidae) perfectly fits the only complete description of that species from the Fiji Islands.

Prevalence of Escherichia coli in surface waters of Southeast Asian cities.

Surface water samples were collected from rivers which fed into large urban areas within Vietnam, Indonesia, Cambodia, and Thailand and were processed to enumerate Escherichia coli. Selected isolates were further characterized using PCR to detect the presence of specific virulence genes. Analyzing the four countries together, the approximate mean cfu/100 ml for E. coli counts in the dry season were log 4.3, while counts in the wet season were log 2.8. Of the 564 E. coli isolates screened for the presence of pathogenic genes, 3.9 % possessed at least one virulence gene. The most common pathogenic types found were Shiga toxin-producing E. coli isolates. These results reinforce the importance of monitoring urban surface waters for fecal contamination, that E. coli in these water environments may serve as opportunistic pathogens, and may help in determining the impact water usage from these rivers have on the public health of urban populations in Southeast Asia.

On four species of echinorhynchid acanthocephalans from marine fish in Halong Bay, Vietnam, including the description of three new species and a key to the species of Gorgorhynchus.

Four species of echinorhynchid acanthocephalans were collected from marine fish off Cat Ba Island, Halong Bay, Gulf of Tonkin, Vietnam, in the spring of 2009. Acanthocephalus halongensis n. sp. (Echinorhynchidae) from the redtail scad, Decapterus kurroides Bleeker 1855 (Carangidae), has a unique proboscis armature with a spiniform basal hook with lateral root and an incomplete receptacle wall posteriorly. Gorgorhynchus tonkinensis n. sp. (Rhadinorhynchidae) also from D. kurroides, has long, slender, winding lemnisci, many epidermal nuclei, and a narrow anterior trunk with a shoulder armed with 20 circles of tightly packed spines, the posterior four circles of which have abruptly larger spines than those in the anterior circles. Neorhadinorhynchus atypicalis n. sp. (Cavisomidae) from the rabbitfish, Siganus fuscescens (Houttuyn 1782) (Siganidae), has the largest number of proboscis hooks per row, testes wider than long, and four clustered cement glands. Micracanthorhynchica kuwaitensis Amin and Sey 1996 (Rhadinorhynchidae) from the spottail needlefish Strongylura strongylura (van Hasselt 1823) (Belonidae) was similar to specimens originally described from the Arabian Gulf off the Kuwaiti coast. These acanthocephalans were collected in small numbers but stood out as uniquely and considerably different from their closest relatives to warrant their reporting. All species of acanthocephalans and their host and geographic distribution are described, and a key to the species of Gorgorhynchus is provided.

On a new acanthocephalan family and a new order, from birds in Vietnam.

During November 2000, a collection of acanthocephalans from birds in Quang Ninh Province, Vietnam, included a new genus and species in a new family and order. Pyrirhynchus heterospinus n. gen., n. sp. (Pyrirhynchidae n. fam.: Heteramorphida: new order) is described from Actitis hypoleucos (Linnaeus, 1758) (common sandpiper). The new family combines characters from Polymorphidae and Heteracanthocephalidae, and it includes new features. Specimens of the new species are distinguished from those of Heteracanthocephalidae and/or Polymorphidae by their long cylindrical trunk with anterior swelling, pyriform proboscis with hooks much larger ventrally, brain at the anterior end of the receptacle, specialized tubular cement glands, and elliptoid eggs with concentric shells. A detailed analysis of proboscis and trunk armature is included, and specimens of several species of Arhythmorhynchus Lühe, 1911 (Polymorphidae) were studied for comparative purposes. The proboscis of P. heterospinus is armed with 17 to 20 rows of 17 to 19 hooks each, with anterior 9-11 hooks rooted and posterior 6-10 spines rootless.

Synthesis and antifungal activities of phenylenedithioureas.

A total of 20 new phenylenedithiourea derivatives was synthesized by reaction of phenylenediisothiocyanates with aromatic amines as aminobenzoic, aminosalicylic acid and their derivatives. Their chemical structures were confirmed by elemental analysis, IR spectrometry and 1H NMR. The compounds were screened for in vitro antifungal, antibacterial activities and some of them have strong antifungal activities comparable to the activity observed for ketoconazole.