Draft genome sequence data of multidrug-resistant Pseudomonas aeruginosa WO7 from a hospital wastewater treatment plant in Thailand.

Multidrug-resistant Pseudomonas aeruginosa WO7 was isolated from an untreated water sample from a hospital wastewater treatment plant in Thailand. This report presents the draft genome sequence data of P. aeruginosa WO7. Genomic DNA was obtained from a pure culture of P. aeruginosa WO7, and paired-end reads were generated using an Illumina MiSeq sequencer. The draft genome consisted of 111 contigs with a total size of 6,784,206 base pairs, an N50 of 209,424 base pairs, and a GC content of 65.85%. The dDDH value between WO7 and Pseudomonas aeruginosa DSM 50071T was determined to be 90.7%, indicating that the strain is Pseudomonas aeruginosa. The data presented indicate the potential for bacterial classification, comparative genomics, comprehensive analysis of antimicrobial resistance, and assessment of bacterial virulence factors in P. aeruginosa. The draft genome sequence data have been deposited at the NCBI under Bioproject accession number PRJNA550309.

Relationship between Potential Trace Elements Contamination in Sediment and Macrofauna in the Upper Gulf of Thailand.

The relationship between heavy metal contamination in sediment and macrofauna in the upper Gulf of Thailand is presented as an indication of the environmental quality of coastal waters. This study aims to monitor five heavy metal elements (As, Cd, Mn, Ni, and Pb) between sediment and the sandworm (Perinereis quatrefagesi (P. quatrefagesi)). The geoaccumulation index (I geo), enrichment factor (EF), and contamination factor (CF), including the pollution load index (PLI), were used for statistical analyses by one-way analysis of variance (ANOVA), and differences in the data were compared using the least significant difference (LSD) test. The sediment heavy metal concentrations were found to decrease (Al > Mn > Pb > Ni > Cd > As), and the quantity of heavy metal contamination in the sediment was not over the emergency point defined by the Pollution Control Department in Thailand. The Mn at the SH and SP points has I geo > 0 but <1 and is considered not polluted to moderately polluted. The EF overall is <2 and is deficient in mineral enrichment. The CF of the Samut Sakhon (SH) and Samut Prakan (SP) sites had high Al and Mn values, indicating a low pollution level, but the PLI had an all-site average of 0.0215 ± 0.0252, which is <1. This indicates that the present areas are close to ideal and not polluted. To measure heavy metals in macrofauna, P. quatrefagesi was assessed, and concentrations were found to decrease (Mn > Ni > Cd > Pb > As). The Mn between sediment and the sandworm was directly correlated (r = 0.976; p < 0.01). The sandworm performs as a bioindicator for the quality of coastal sediment, specifically with Mn; thus, the result present at a baseline level may grow in the future if there is no control measure for defensive measures.

Molecular assessment and transcriptome profiling of wild fish populations of Oryzias mekongensis and O. songkhramensis (Adrianichthyidae: Beloniformes) from Thailand.

Among the fish of the genus Oryzias, two species are frequently used as model animals in biological research. In Thailand, Oryzias mekongensis is usually found in natural freshwater near the Mekong Basin in the northeast region, while O. songkhramensis inhabits the Songkhram Basin. For differential morphological identification, the coloured bands on the dorsal and ventral margins of the caudal fin are used to distinguish O. mekongensis from O. songkhramensis. However, these characteristics are insufficient to justify species differentiation, and little molecular evidence is available to supplement them. This study aimed to investigate the molecular population and transcriptome profiles of adult O. mekongensis and O. songkhramensis. In the molecular tree based on cytochrome b sequences, O. mekongensis exhibited four clades that were clearly distinguished from O. songkhramensis. Clade 1 of the O. mekongensis population was close to the Mekong River and lived in the eastern portion of the upper northeast region. Clade 2 was far from the Mekong River and inhabited the middle region of the Songkhram River. Clade 3 was positioned to the west of the Songkhram River, and clade 4 was to the south of the Songkhram River Basin. After RNA sequencing using an Illumina HiSeq 2500 platform, the gene category annotations hardly differentiated the species and were discussed in the text. Based on the present findings, population dispersal of these Oryzias species might be associated with geographic variations of the upper northeast region. Molecular genetics and transcriptome profiling might advance our understanding of the evolution of teleost fish.

Molecular analysis of population and De Novo transcriptome sequencing of Thai medaka, Oryzias minutillus (Teleostei: Adrianichthyidae).

Thai medaka (Oryzias minutillus) are alternatively known as Thai rice-fish or dwarf medaka, and they widely inhabit natural freshwater environments in all regions of Thailand. In this study, we aimed to investigate the molecular genetics of the Thai medaka population in Thailand inferred from the mitochondrial control region (D-loop) and the cytochrome c oxidase subunit 1 (coxI) sequences. Furthermore, we examined RNA sequencing (RNA-seq) of adult males and females was performed with next-generation sequencing. Together, the combination of the D-loop and coxI sequences clearly distinguished the Thai medaka populations into 2 groups, such as group 1, which generally included samples from the central, northern, western, and eastern regions of the northeastern region. In this group, the fish populations seem to be a little monophyly in which the first subpopulation comprised the main samples from the northern and central regions. The second subpopulation commonly contained fish from the eastern region and specimens from the southern part of the central region near the Gulf of Thailand. Although these subgroups related to geographical distribution, bootstrap values were low in branch considered significant for both subgroups. Group 2 consisted of almost all samples from the southern population and those from the central and southern part of the northeastern region. Group 2 was found that it was made of samples from the northeastern region and samples from the southern population. A total of 73551 unigenes were identified after gene annotation. Signal transduction was the predominant protein classification among the Thai medaka orthologous groups. A differentially expressed gene (DEG) analysis identified 6 subclusters between both sexes that were composed of 257, 131, 364, 386, 114 and 108 genes. Phototransduction was the most enriched pathway and was highly expressed in males, while viral carcinogenesis, oocyte genesis, and the complement and coagulation cascades were highly expressed in females. Further details of these DEGs are discussed below. These results suggest that Thai medaka may genetically exhibit independent populations in the geographic habitats of Thailand. Moreover, these fish also reveal the genes that are conserved in other organisms and those that may be specific to this species.

Contributions to the knowledge of Pseudolevinseniella (Trematoda: Digenea) and temnocephalans from alien crayfish in natural freshwaters of Thailand.

Redclaw crayfish (Cherax quadricarinatus) is a decapod species originating from Australian freshwater. For more than two decades, these crayfish have been re-distributing to environments in many countries, including Thailand. Moreover, they can carry endosymbionts and/or ectosymbionts into new environments. The aim of this study was to introduce a morphological description of Pseudolevinseniella anenteron as a metacercaria of the endoparasites of redclaw crayfish collected from natural water sources in Thailand. The occurrence of two ectosymbiotic temnocephalans (Diceratocephala boschmai and Temnosewellia sp.) in C. quadricarinatus was also reported. The internal morphology of P. anenteron, D. boschmai and Temnosewellia were described and discussed. The surface ultrastructure of the multidentate spines on the body and the metacercarial cyst wall of P. anenteron was investigated by scanning electron microscopy (SEM). By performing a search of the GenBank nucleotide database of partial sequences of 18S, 28S rDNA and cytochrome c oxidase subunit I (cox1), P. anenteron was found to be related to Maritrema, and Temnosewellia was found to be related to T. fasciata. However, according to the cox1 gene, Temnosewellia was found to be similar to T. minor. These results reveal that redclaw crayfish that inhabit natural freshwaters in Thailand may harbour endoparasites and ecto- and endosymbionts. Furthermore, these findings may be able to monitor invasive or non-invasive species in an ecosystem.

The Anthelmintic Effects of Medicinal Plant Extracts Against Paramphistome Parasites, Carmyerius spatiosus.

INTRODUCTION: Paramphistomosis is a disease caused by the rumen flukes which cause an acute gastroenteritis and anemia with high mortality particularly in young ruminants. MATERIALS AND METHODS: In this study, we have investigated the anthelmintic effect of medicinal plant extracts from leaves and heartwoods of Cassia siamea L., roots of Plumbago zeylanica L. and Plumbago indica L., and leaves of Terminalia catappa L. against Carmyerius spatiosus. RESULTS: The highest anthelminthic effect on the flukes after 24 h of exposure was found in heartwood ethyl acetate extract of C. siamea (LC50 = 374.30; LC90 = 749.03 ppm), root n-butanol extract of P. zeylanica (LC50 = 1005.12; LC90 = 2411.55 ppm), root hexane, ethyl acetate, and n-butanol extract of P. indica (LC50 = 34.38, 211.34, 506.92; LC90 = 64.09, 496.05, 934.86 ppm), and leaf n-butanol and water extract of T. catappa (LC50 = 487.17, 470.28; LC90 = 913.27, 848.23 ppm). When observed by scanning electron microscopy, the tegument showed similar sequence of morphological changes after treatments with all plant extracts, comprising of swelling of ridges and folds, blebbing, rupturing of the blebs, erosion, lesion and disruption of the tegument. CONCLUSION: This study is the first report on the anthelmintic activity of plant extracts to C. spatiosus; therefore, these plant extracts are highly effective in the elimination of adult rumen flukes.

The in vitro anthelmintic activity of the ethanol leaf extracts of Terminalia catappa L. on Fasciola gigantica.

At present, there are no medicinal plant extracts currently available for treatment and control of fasciolosis. The present work could provide, for the first study, conclusions on the in vitro fasciolicidal properties of the ethanol extract of Terminalia catappa L. (TcCE) leaves against adult Fasciola gigantica after incubation with RPMI-1640 medium containing the TcCE at various concentrations and times when compared with triclabendazole (TCZ). The relative motility and survival index values of the TcCE-treated flukes decreased at a more rapid rate than the TCZ-treated flukes. The death of the parasites was observed after exposed to TcCE at 3 h incubation with 400, 800 and 1000 µg mL-1, and at 6 h incubation in 100 and 200 µg mL-1. Vacuolization, blebbings and partial disruption on the parasites' tegument were observed by light microscopy. When examined by scanning electron microscopy, TcCE caused similar tegumental alterations in the parasites as those observed in TCZ treatment but with larger damage at comparative incubation periods, consisting of swelling, blebbing, disrupted blebs, loss of spines, leading to the erosion, lesion and eventual disruption of the total tegument. Therefore, the TcCE may exert its fasciolicidal effect against F. gigantica by initially causing the tegumental alteration.

The anthelmintic effects of the ethanol extract of Terminalia catappa L. leaves against the ruminant gut parasite, Fischoederius cobboldi.

Presently, no effective anthelmintic drugs have been used to treat and control paramphistomosis, a severe disease of ruminants. In this study, we have investigated the in vitro anthelmintic effect of the leaves of Terminalia catappa L. crude extract (TcCE) and albendazole (ABZ) on adult Fischoederius cobboldi after incubating the flukes in RPMI-1640 medium containing the TcCE at various doses and times. The TcCE-treated flukes at all dosages exhibited rapid decrease of motility, and the relative motility (RM) values were decreased sharply from start to 3 h. Worms were killed after 6 and 12 h of treatment with 1000, 1500 and 2000 µg mL(-1) as well as 500 µg mL(-1) of TcCE, respectively. By light microscopy examination, the flukes exhibited the earliest alteration in a limited area of the tegument. At scanning electron microscopy level, the flukes' tegument showed similar sequence of morphological alterations after treatment with ABZ and TcCE that consisted of swelling of ridges and folds, followed by blebbing and rupturing of the blebs, leading to the erosion, lesion and disruption of the tegument. Hence, in vivo studies should be performed to examine whether the TcCE may serve as a powerful anthelmintic drug for treatment of paramphistomosis.

Surface topography and ultrastructural architecture of the tegument of adult Carmyerius spatiosus Brandes, 1898.

Adult Carmyerius spatiosus or stomach fluke has an elongate, cylindrical-shaped, straight to slightly curved body, with conical anterior end and truncated posterior end. The worm measures about 8.7-11.2mm in body length and 2.3-3.0mm in body width across the mid-section. When observed by SEM, the tegumental surface in all part of the body appears highly corrugated with ridges and furrows, and having no spines. The ventral surface has more complex corrugation than those of the dorsal surface. Both anterior and posterior suckers have thick edges covered with transverse folds and appear spineless. The genital pore is located at the anterior part of the body. There are two types of sensory papillae on the surface: type 1 is bulbous in shape with nipple-like tips; type 2 has a similar shape with short cilia on the tip. The dorsal surface exhibits similar surface features, but papillae appear less numerous and are smaller. When observed by TEM, the tegument is divided into four layers. The first layer includes the ridges and furrows which are covered by a trilaminate membrane underlined by a dense lamina and coated externally with the glycocalyx. The second layer of the tegument is a narrow region of cytoplasm that contains high concentrations of ovoid electron lucent tegumental granules (TG1), and disc-shaped electron dense tegumental granules (TG2) as well as lysosomes. TG1 close to the surface invariably exocytose their content into bottoms of the ridges, while some TG2 are fused and have their membrane joined up with the surface membrane. The third layer is the widest middle area of the tegument which contains numerous and evenly distributed mitochondria. Both TG1 and TG2 granules are present but in much fewer number than in the first and second layers. The fourth layer is the innermost zone that rests on and couples with a thick basal lamina. The cytoplasm in this layer is loosely packed and contains numerous infoldings of the basal plasma membrane with closely associated mitochondria. It also contains fairly large numbers of TG1 and TG2 granules which are produced and transported to the tegument by one type of tegumental cells lying in rows underneath the muscular layers.

Morphology and histology of the adult Paramphistomum gracile Fischoeder, 1901.

In the present study, we evaluated the histological morphology of the adult Paramphistomum (P.) gracile. Adult flukes with bodies 5~15 mm in length and 2~7 mm in width were subjected to histological analysis. Longitudinal and transversal serial-sections were stained with hematoxylin and eosin, and examined. The body surface and longitudinal section of P. gracile were also assessed using scanning electron microscopy. In this species, the anterior sucker and posterior sucker (acetabulum) were present on an anterior and posterior part of the body, respectively. The major folds were located in the areas of the anterior sucker, genital canal, and posterior sucker. The fluke membrane was spineless at the tegument surface and in the tegument tissue. Histological data showed structural-systematic characteristics of the digestive tract, reproductive tract, excretory tract, copulatory organs, connective tissues, and muscle tissues. We attempted to elucidate the histological characteristics of P. gracile that might increase the knowledge and understanding of rumen fluke morphology.

Fischoederius cobboldi: A scanning electron microscopy investigation of surface morphology of adult rumen fluke.

Adults Fischoederius cobboldi are conical-shaped, concave ventrally and convex dorsally, measures about 8-10mm in length and 4-6mm in width across the mid section. Scanning electron microscopy (SEM) of entire body showed that the tegumental surface exhibits highly corrugation and transverse folds alternating with grooves and without spines. At higher magnification, the surface of each fold is further increased with a meshwork of ridges separated by irregular-sized pits. The ventral surface has more complex corrugations and invaginations than those of the dorsal surface of the body. Both anterior and posterior suckers have thick edges covered with transverse folds and appear spineless. The genital pore is located at the anterior one-third of the body. There are two types of sensory papillae on the surface: type 1 is bulbous in shape and nipple-like tips, measuring 10-15 µm in diameter at the base, and also type 2 is a similar shape and has short cilia on tips. These sensory papillae occur in large clusters, each having between 7 and 25 units depending on the region of the body. Clusters of papillae on the ventral surface and around the anterior suckers tend to be more abundant and larger in size. The dorsal side of the body exhibit similar surface features, but papillae appear less numerous and are smaller. Corrugations and invaginations of the dorsal aspect are also less extensive than those on the ventral surface of the body.

Expression levels of hormone receptors and bone morphogenic protein in fins of medaka.

In the genus Oryzias, the morphologies of the dorsal and anal fins are typical secondary sex characters. In the Japanese medaka (Oryzias latipes) and Thai medaka (Oryzias minutillus), androgen receptor (AR) expression levels in the dorsal, anal, and pectoral fins were higher in males than in females. Conversely, in both species estrogen receptor (ER) beta expression levels in the dorsal and anal fins were higher in females than in males. AR and ERbeta expression levels in the dorsal and anal fins of sex-undeterminable individuals of Thai medaka were intermediate between those in normal male and female Thai medaka. There was no difference in the bone morphogenic protein (Bmp) 2b expression level between male and female Japanese medaka. In contrast, the Bmp2b expression level in the dorsal fin of sex-undeterminable individuals was lower than in normal male and female Thai medaka. It is thus clear that androgen and estrogen regulate the sex-dependent characters of fin morphology in both Oryzias species. In sex-undeterminable individuals of Thai medaka, the low levels of Bmp2b expression in the dorsal fin are evidence that androgen and estrogen are necessary for adequate expression of Bmp2b in the normal development of at least the dorsal fin.