In vitro cytotoxicity of nonpolar constituents from different parts of kava plant (Piper methysticum)

Kava (Piper methysticum), a perennial shrub native to the South Pacific islands, has been used to relieve anxiety. Recently, several cases of severe hepatotoxicity have been reported from the consumption of dietary supplements containing kava. It is unclear whether the kava constituents, kavalactones, are responsible for the associated hepatotoxicity. To investigate the key components responsible for the liver toxicity, bioassay-guided fractionation was carried out in this study. Kava roots, leaves, and stem peelings were extracted with methanol, and the resulting residues were subjected to partition with a different polarity of solvents (hexane, ethyl acetate, n-butanol, and water) for evaluation of their cytotoxicity on HepG2 cells based on the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase and aspartate aminotransferase enzyme leakage assays. Organic solvent fractions displayed a much stronger cytotoxicity than water fractions for all parts of kava. The hexane fraction of the root exhibited stronger cytotoxic effects than fractions of root extracted with other solvents or extracts from the other parts of kava. Further investigations using bioassay-directed isolation and analysis of the hexane fraction indicated that the compound responsible for the cytotoxicity was flavokavain B. The identity of the compound was confirmed by (1)H and (13) C NMR and MS techniques.     

To pool or not to pool? Guidelines for pooling samples for use in surveillance testing of infectious diseases in aquatic animals

Samples from multiple animals may be pooled and tested to reduce costs of surveillance for infectious agents in aquatic animal populations. The primary advantage of pooling is increased population‐level coverage when prevalence is low (<10%) and the number of tests is fixed, because of increased likelihood of including target analyte from at least one infected animal in a tested pool. Important questions and a priori design considerations need to be addressed. Unfortunately, pooling recommendations in disease‐specific chapters of the 2018 OIE Aquatic Manual are incomplete and, except for amphibian chytrid fungus, are not supported by peer‐reviewed research. A systematic review identified only 12 peer‐reviewed aquatic diagnostic accuracy and surveillance studies using pooled samples. No clear patterns for pooling methods and characteristics were evident across reviewed studies, although most authors agreed there is a negative effect on detection. Therefore, our purpose was to review pooling procedures used in published aquatic infectious disease research, present evidence‐based guidelines, and provide simulated data examples for white spot syndrome virus in shrimp. A decision tree of pooling guidelines was developed for use by peer‐reviewed journals and research institutions for the design, statistical analysis and reporting of comparative accuracy studies of individual and pooled tests for surveillance purposes.     

Design standards for experimental and field studies to evaluate diagnostic accuracy of tests for infectious diseases in aquatic animals

Design and reporting quality of diagnostic accuracy studies (DAS) are important metrics for assessing utility of tests used in animal and human health. Following standards for designing DAS will assist in appropriate test selection for specific testing purposes and minimize the risk of reporting biased sensitivity and specificity estimates. To examine the benefits of recommending standards, design information from published DAS literature was assessed for 10 finfish, seven mollusc, nine crustacean and two amphibian diseases listed in the 2017 OIE Manual of Diagnostic Tests for Aquatic Animals. Of the 56 DAS identified, 41 were based on field testing, eight on experimental challenge studies and seven on both. Also, we adapted human and terrestrial‐animal standards and guidelines for DAS structure for use in aquatic animal diagnostic research. Through this process, we identified and addressed important metrics for consideration at the design phase: study purpose, targeted disease state, selection of appropriate samples and specimens, laboratory analytical methods, statistical methods and data interpretation. These recommended design standards for DAS are presented as a checklist including risk‐of‐failure points and actions to mitigate bias at each critical step. Adherence to standards when designing DAS will also facilitate future systematic review and meta‐analyses of DAS research literature.     

Development and application of molecular methods (PCR) for detection of Tasmanian Atlantic salmon reovirus

Molecular (PCR) diagnostic tests for the detection and identification of aquareovirus in general, and Tasmanian Atlantic salmon reovirus (TSRV) specifically, were developed, and their diagnostic sensitivity and specificity were determined and compared with virus isolation in cell culture. Intralaboratory and interlaboratory comparison of PCR (conventional hemi‐nested RT‐PCR & RT‐qPCR) and virus isolation in cell culture using finfish cell lines, CHSE‐214 and EPC, was carried out for the detection and identification of TSRV using field samples of farmed Atlantic salmon Salmo salar, L. from various aquaculture sites around Tasmania. The interlaboratory comparison of diagnostic methods was carried out between two laboratories, AAHL‐CSIRO and DPIPWE‐Tasmania. A total of 144 fish from nine sites (12–33 fish per site) were sampled from two regions of Tasmania (Tamar River estuary in the north and Huon River estuary in the south‐east) during late spring to early summer of 2009, and the data were analysed using different statistical approaches. The prevalence of TSRV ranged from 6% to 22% in both regions. All the diagnostic methods (data from both laboratories) had high specificity, while the estimated sensitivity varied between tests with RT‐qPCR being the most sensitive (95.2%) method followed by virus isolation and then conventional hemi‐nested RT‐PCR.     

Dissemination of weeds with rice seedlings

Abstract

Dissemination of weed seeds, weed seedlings, and shoots of perennial weeds along with transplanted rice (Oryza sativa L.) seedlings was observed in farmers’ fields in Guimba, Nueva Ecija, Philippines. The percentage of weed seedlings and shoots in bundles of rice seedlings to be transplanted into the field ranged from 4–5 to 14.2% in the wet season and from 3.0 to 8.7% in the dry season. Grasses comprised 63% of the weed seedlings in the wet season and 81% in the dry season. The number of weed seeds disseminated with the rice seedlings ranged from 271 to 683 per bundle. Mainly sedges and broadleaf weeds were disseminated in this manner. Possible ways of reducing dissemination of weeds with rice seedlings are suggested.     

Influence of genetic type, slaughter weight and sex on ovine muscle fiber and fat-cell development

Histological properties from the longissimus (LD) and semimembranosus (SM) muscles of 51 wether and ewe lambs from Hampshire rams and two ewe genetic types (SR, 1/2 Suffolk and 1/2 Rambouillet, and FS, 1/2 Finnish Landrace and 1/2 Southdown) and three slaughter live weights (32, 41 and 50 kg) were compared. Fibers in both muscles were classified as beta R (red), alpha R (intermediate) or alpha W (white). All LD muscle fiber types from FS ewe lambs increased in diameter from 32 to 41 kg, but decreased in diameter from 41 to 50 kg. Also, this quadratic effect with slaughter weight was found in alpha R and alpha W fibers from the SM muscle of FS wether and ewe lambs. However, diameters of alpha R and alpha W fibers from SR wether and ewe lambs and beta R fibers from SR ewe lambs increased linearly in the SM muscle with increasing slaughter weight. As slaughter weight increased, the proportion of alpha R fibers decreased in both the LD and SM muscles for SR wether and FS wether and ewe lambs. Concurrent with the increase in slaughter weight and decrease in alpha R fiber percentage, the proportion of alpha W fibers increased in the LD muscle of SR wether lambs, the SM muscle of SR ewe lambs and both muscles of FS wether and ewe lambs. Genetic type and sex groups, except SR ewe lambs, support the theory of transformation of alpha R to alpha W fibers with increasing slaughter weights. Fat-cell diameters increased in both muscles of SR wether and FS ewe lambs with increasing slaughter weights. Increases in fat traits of lamb carcasses were related to increases in red-fiber size. Alpha red fiber numbers were inversely related (P less than .05) to alpha W fiber numbers in both the LD and SM muscles (r = -.83 and -.79). The proportion of alpha R to alpha W fibers might be used as an indicator of physiological maturity for lambs.     

The effect of 1,8-naphthalic anhydride and depth of seeding on rice tolerance to herbicides in greenhouse tests

In greenhouse tests both butachlor and thiobencarb injured the rice (Oryza sativa L.) cultivars IR36 and Salumpikit. Depth of seeding altered the tolerance of both cultivars to the herbicides. Planting IR36 3 cm deep and Salumpikit 4 cm deep protected them from herbicide damage. Planting deeper than 4 cm generally resulted in smaller and lighter plants. Seed treatment with 1,8-naphthalic anhydride (NA) at 1% w/w further reduced the phytotoxic effect of the herbicides, especially at shallower planting depths. The plants from the NA-treated seeds were taller and heavier than those from the untreated seeds.