Toxicity of essential oil compounds against diamondback moth,Plutella xylostella,and their impact on detoxification enzyme activities

Essential oils are by-products of plant metabolism that are now known to interfere with basic metabolic, physiological and behavioral functions of insects, thereby having promise for use as pest control agents. Accordingly, four essential oil compounds, thymol, 1,8-cineole, linalool and pulegone, were evaluated against the diamondback moth, Plutella xylostella (Linnaeus) (Lepidoptera: Yponomeutidae), to determine their acute toxicity. Thymol and 1,8-cineole were highly toxic to third instars with a LD₅₀ of 0.22 and 0.41 μg/larva, respectively. Linalool and pulegone were moderately active against this insect species, exhibiting <45 % mortality in up to 15 μg/larva. However, in biorational mixtures pulegone was synergistic to both thymol and 1,8-cineole where the increase in activity was almost twofold. Thymol and 1,8-cineole, though active individually against P. xylostella larvae, were antagonistic as biorational mixtures. Linalool was antagonistic in all combinations. In various assays with detoxification enzymes in treated conditions, there was a significant increase in enzyme levels both in vivo and in vitro. Thymol and 1,8-cineole were the active toxicants against P. xylostella, with significant potential to control this pest as biorational mixtures in a synergistic combination with pulegone. Induction in enzyme levels by these compounds suggests possibilities of resistance, which at present remains a speculation, but cannot be summarily ignored because the induction of enzymes due to involvement of detoxification enzymes in the metabolism of a broad range of xenobiotics and secondary metabolites in plants is well known.     

The effects of manipulation medium, culture system and recipient cytoplast on in vitro development of intraspecies and intergeneric felid embryos

The aim of this study was to investigate if reconstructed felid embryos obtained by intraspecies or intergeneric cloning can develop in vitro. Fibroblast cells (f) from a domestic cat (DCf), marbled cat (MCf) and bovine (Bf) were used as donor cells, and oocytes (o) from domestic cats (DCo) and bovine (Bo) were used as recipient cytoplasts. There were two intraspecies (donor cell + recipient cytoplast: DCf + DCo and Bf + Bo) and three intergeneric (MCf + DCo, DCf + Bo and MCf + Bo) cloning groups in the study. In Experiment 1, the effects of manipulation media, modified TCM-199 (199H) or Emcare holding medium (EHM), on in vitro development of DCf + DCo embryos were investigated. The blastocyst formation rate (BFR) of the embryos manipulated in EHM (33.3%) was higher (P<0.05) compared with those manipulated in 199H (18.1%). In Experiment 2, DCf + DCo and MCf + DCo embryos were cocultured with or without domestic cat oviductal epithelium cells. Irrespective of coculture, the same BFR was obtained for DCf + DCo embryos (44.4 vs. 38.0%), while MCf + DCo embryos could not develop beyond the morula stage. In experiment 3, although the development of MCf + DCo and DCf + Bo embryos was arrested at the morula stage, 8.6% of MCf + Bo embryos were able to develop to the blastocyst stage. These results demonstrated that EHM was superior to 199H as an embryo manipulation medium and that the DCo and Bo could support the early embryonic development of intergeneric cloned marbled cat embryos up to the morula stage. However, postimplantation development still needs to be investigated.     

The insecticidal potential of Piper ribesioides (Piperales: Piperaceae) extracts and isolated allelochemicals and their impact on the detoxification enzymes of Spodoptera exigua (Lepidoptera: Noctuidae)

Phytoparasitica - The dried twigs of Piper ribesioides were used to prepare various extracts in different solvents. Each extract was evaluated for their toxicity against 2nd instar larvae of...     

In vitro assembly of Penaeus monodon densovirus (PmDNV)‐like particles produced in a prokaryote expression system

Viral diseases are a significant problem in the shrimp aquaculture industry as outbreaks can cause significant mortality and economic loss. While it has been shown that triggering the shrimp RNA interference pathway through dsRNA is a potentially viable treatment pathway, this approach is hampered by the lack of a suitable delivery mechanism. Virus‐like particles (VLPs), which are structurally similar to native viruses but lack the genetic material, could possibly be developed as a delivery vehicle. To generate a candidate VLP, the Penaeus monodon densovirus (PmDNV) capsid protein was cloned with an added histidine tag and expressed in an E. coli expression system. While the protein was expressed in inclusion bodies, the recombinant PmDNV capsid protein could be dissolved and subsequently purified by nickel affinity column chromatography. The formation of VLP from this purified rPmDNV capsid protein was investigated by transmission electron microscopy, and PmDNV‐VLPs were observed that looked similar to the native PmDNV virion. Our results suggest that the PmDNV‐like particle could be promisingly applied towards vaccination and that this PmDNV‐like particle can potentially serve as a system for delivery of nucleic acids to trigger innate immunity in shrimp.     

Culture-independent analysis of the bacterial community during fermentation of pa-som,a traditional fermented fish product in Laos

Pa-som is a traditional salty fermented food made from varieties of freshwater fish with garlic and rice in Laos. A similar product, plaa-som, is made in Thailand. To investigate the fermentation process of pa-som, lactic acid production and pH were monitored during laboratory-scale production of pa-som. Furthermore, the variety of bacteria involved and their succession during fermentation were examined using culture-independent polymerase chain reaction (PCR) denaturing gel gradient electrophoresis targeting the 16S ribosomal RNA (rRNA) gene. d- and l-lactic acid production and a decrease in pH were observed in the samples after 1 day of fermentation. The lactic acid content and pH continued to increase and decrease, respectively, until 4 days of fermentation. Overall, six lactic acid bacteria and eight other indigenous bacteria species were detected during pa-som fermentation. Among the lactic acid bacteria detected, two Lactococcus and one Weissella species were detected in samples after 1–4 days of fermentation, suggesting that these three species play major roles in pa-som fermentation from the initial phase. This is the first report to investigate the fermentation process in pa-som at the microbial level. This research approach should be applicable to a wide variety of pa-som and related fermented fishery products in Laos to help understand their microbial diversity and to identify beneficial bacterial species for improving quality.     

Differential detection of Trichinella papuae, T. spiralis and T. pseudospiralis by real-time fluorescence resonance energy transfer PCR and melting curve analysis

Trichinellosis caused by nematodes of Trichinella spp. is a zoonotic foodborne disease. Three Trichinella species of the parasite including Trichinella spiralis, Trichinella papuae and Trichinella pseudospiralis, have been etiologic agents of human trichinellosis in Thailand. Definite diagnosis of this helminthiasis is based on a finding of the Trichinella larva (e) in a muscle biopsy. The parasite species or genotype can be determined using molecular methods, e.g., polymerase chain reaction (PCR). This study has utilized real-time fluorescence resonance energy transfer PCR (real-time FRET PCR) and a melting curve analysis for the differential diagnosis of trichinellosis. Three common Trichinella species in Thailand were studied using one set of primers and fluorophore-labeled hybridization probes specific for the small subunit of the mitochondrial ribosomal RNA gene. Using fewer than 35 cycles as the cut-off for positivity and using different melting temperatures (T(m)), this assay detected T. spiralis, T. papuae and T. pseudospiralis in muscle tissue and found the mean T(m) ± SD values to be 51.79 ± 0.06, 66.09 ± 0.46 and 51.46 ± 0.09, respectively. The analytical sensitivity of the technique enabled the detection of a single Trichinella larva of each species, and the detection limit for the target DNA sequence was 16 copies of positive control plasmid. A test of the technique's analytical specificity showed no fluorescence signal for a panel of 19 non-Trichinella parasites or for human and mouse genomic DNA. Due to the sensitivity and specificity of the detection of these Trichinella species, as well as the fast and high-throughput nature of these tools, this method has application potential in differentiating non-encapsulated larvae of T. papuae from T. spiralis and T. pseudospiralis in tissues of infected humans and animals.     

Highly efficient double‐stranded RNA transfection of penaeid shrimp using cationic liposomes

Yellow head virus (YHV) is a viral pathogen of shrimp that has caused high economic loss in the Thai shrimp farming industry, and the RNA interference mechanism has potential as a virus control strategy, as previous studies have shown that injection of virus specific double‐stranded RNA (dsRNA) can inhibit viral replication in shrimp. However, to date, complete inhibition viral replication has not been achieved. This study sought to determine whether cationic liposomes mixed with an antiviral dsRNA (YHV‐Pro‐dsRNA) were able to enhance protection against YHV. The results showed that injection of a YHV‐Pro‐dsRNA/cationic liposome complex markedly increased protection against YHV as compared with naked YHV‐Pro‐dsRNA, suggesting that cationic liposomes enhance the uptake of protective dsRNA into shrimp cells.     

Urban green space, street tree and heritage large tree assessment in Bangkok, Thailand

Three inventories were conducted to quantify Bangkok's green infrastructure for future planning and improvement in the context of a seasonal monsoonal dry climate. Total green space was quantified by extracting surface cover areas from remotely sensed data in a geographical information system (GIS) environment, and this information was used to designate suitable sites for future green spaces such as parks. Street trees were inventoried for species identification and size. Trees of heritage value were identified through a public awareness campaign, and then were subsequently surveyed for species identification, height and trunk diameter. GIS green space analysis showed that per capita park space was approximately 1.8 m², but a master plan proposed increasing per capita park area to 4 m² within 25 years. The increased park area will be largely in the form of lower cost, semi-naturalized tree parks. The inventoried street tree population, approximately 200,000, was skewed somewhat towards a monoculture, as 42% were the facultative evergreen Pterocarpus indicus Wild. By contrast, none of the other species exceeded 7% of the total. That most of the other species consisted of smaller trunk diameters than P. indicus, and therefore younger, suggests that Bangkok's street tree plantings are becoming more diverse. The heritage large tree inventory was dominated by evergreen tree species, particularly exceptionally large Ficus species, found largely on Buddhist temple grounds, followed by Albizia saman (Jacq.) Merr. The slower growing evergreen heritage species are worth careful appraisal and preservation because they are less likely to be commonly planted. Careful species selection balancing drought deciduous and dry evergreen species can achieve adaptation to the monsoonal dry season with diverse aesthetic quality in both Bangkok's street tree population and in its semi-naturalized tree parks.     

Seroprevalence of Toxoplasma gondii in captive felids in Thailand

The prevalence of antibodies against Toxoplasma gondii was investigated by commercial latex agglutination test kit (Toxocheck-MT 'Eiken') in captive felids maintained at zoos and a wildlife breeding center in different geographic regions of Thailand. Sera from a total of 136 captive felids of 12 species was obtained between 2002 and 2004. The overall seroprevalence of T. gondii was found in 21 of 136 (15.4%) felids. The titers varied from 1:64 (eight samples) to 1:8192 (one sample). The seroprevalence in different geographic regions were from 0% in the northern area to 23% in the southern area. This study suggested a widespread exposure of captive felids to T. gondii in Thailand and this is the first report of serologic analysis for T. gondii in captive felids in Southeast Asia.