Development and application of polymerase chain reaction-based assays for Rathayibacter Toxicus and a Bacteriophage associated with annual ryegrass (Lolium Rigidum) toxicity

Annual ryegrass toxicity (ARGT) is responsible for significant stock losses in South Australia and Western Australia. The toxicity is caused by corynetoxins produced by the bacterium Rathayibacter toxicus (with the possible involvement of a bacteriophage), which infects annual ryegrass (Lolium rigidum). Polymerase chain reaction (PCR)-based assays, compatible with an existing enzyme-linked immunosorbent assay for the corynetoxins, have been developed and used to screen L. rigidum for both the presence of R. toxicus and for the bacteriophage isolate NCPPB 3778. The results from analysing bacterially infected galls from toxic grain screenings showed a positive correlation between the presence of the bacterium and corynetoxins but not with the bacteriophage. Analysis of pasture-derived samples of annual ryegrass showed about a 50% correlation of corynetoxins with bacterial presence and about a 5% correlation of phage with the presence of the bacterium. These observations support the potential application of the PCR-based assays in providing a useful, complementary tool in the assessment of the likelihood of pasture and feed to cause ARGT and to enable a better understanding of the complex aetiology of ARGT.

Some aspects of photosynthetic characteristics in a set of perennial irrigation reservoirs located in five river basins in Sri Lanka

Phytoplankton primary productivity of eleven irrigation reservoirs located in five river basins in Sri Lanka was determined on a single occasion together with light climate and nutrient concentrations. Although area-based gross primary productivity (1.43–11.65 g O2 m−2 d−1) falls within the range already established for tropical water bodies, net daily rate was negative in three water bodies. Light-saturated optimum rates were found in water bodies, with relatively high algal biomass, but photosynthetic efficiency or specific rates were higher in water bodies with low algal biomass, indicating nutrient limitation or physiological adaptation of phytoplankton. Concentrations of micronutrients and algal biomass in the reservoirs are largely altered by high flushing rate resulting from irrigation release. Underwater light climate and nutrient availability control the rate of photosynthesis and subsequent areabased primary production to a great extent. However, morpho-edephic index or euphotic algal biomass in the most productive stratum of the water column is not a good predictor of photosynthetic capacity or daily rate of primary production of these shallow tropical irrigation reservoirs.

Limnology and culture-based fisheries in non-perennial reservoirs in Sri Lanka

This study was carried out to investigate the possibility of using the limnological characteristics of non-perennial reservoirs in Sri Lanka for the future management of culture-based fisheries. Forty-five reservoirs were randomly selected to study their limnology, out of which 32 were stocked with fish fingerlings of Chinese and Indian carps, tilapia and freshwater prawn at stocking densities ranging from 218–4372 fingerlings ha−1. Of these, 23 reservoirs were harvested at the end of the culture period (6–10 months). Thirteen limnological parameters were measured during the water retention period of each of the 45 reservoirs between November 2001 and January 2004. The mean values of the limnological parameters were used to ordinate the reservoirs through principal component analysis. Ordination showed a productivity gradient among reservoirs where Secchi disc depth, total phosphorus, chlorophyll-a, inorganic turbidity and organic turbidity were identified as key factors. The total fish yield of culture-based fisheries was positively correlated to the scores of the first principal component axis. This study reveals that it is possible to classify non-perennial reservoirs in Sri Lanka based on the above limnological parameters in order to develop culture-based fisheries and that they could be applicable in comparable water bodies elsewhere in the tropics.

Trophic classification of non-perennial reservoirs utilized for the development of culture-based fisheries, Sri Lanka

The aim of this study was to check the suitability of some trophic models developed for temperate regions to classify the non-perennial reservoirs of Sri Lanka in order to manage culture-based fisheries of those reservoirs. A limnological study was carried out in 45 non-perennial reservoirs, which have been randomly selected for stocking of fish fingerlings for the development of culture-based fisheries. High total phosphorous (TP) content in relation to algal biomass indicates high non-algal turbidity in all reservoirs. Carlson's trophic state indices (TSI) measured on the basis of Secchi disc depth [TSI (SDD)], TP [TSI (TP)] and chlorophyll a [TSI (Chl-a)] show that the 45 reservoirs studied are characterized by TSI (TP) = TSI (SDD) > TSI (Chl-a), indicating that non-algal particulate matter or colour dominates underwater light attenuation. As TSI (Chl-a) is positively correlated to culture-based fisheries yield, it is useful for planning culture-based fisheries development strategies in non-perennial reservoirs of Sri Lanka, and has the potential to be used elsewhere in the tropics for comparable developments.

Culture-based fisheries in non-perennial reservoirs in Sri Lanka: production and relative performance of stocked species

In Sri Lanka, there is a great potential for the development of culture-based fisheries because of the availability of around 12 000 non-perennial reservoirs in the dry zone (<187 cm annual rainfall) of the island. These reservoirs fill during the north-east monsoonal period in October to December and almost completely dry up during August to October. As these non-perennial reservoirs are highly productive, hatchery-reared fish fingerlings can be stocked to develop culture-based fisheries during the water retention period of 7–9 months. The present study was conducted in 32 non-perennial reservoirs in five administrative districts in Sri Lanka. These reservoirs were stocked with fingerlings of Indian (catla Catla catla Hamilton and rohu Labeo rohita Hamilton) and Chinese (bighead carp Aristichthys nobilis Richardson) major carps, common carp Cyprinus carpio L., genetically improved farmed tilapia (GIFT) strain of Nile tilapia, Oreochromis niloticus (L.) and post-larvae of giant freshwater prawn, Macrobrachium rosenbergii De Man, at three different species combinations and overall stocking densities (SD) ranging from 218 to 3902 fingerlings ha⁻¹, during the 2002–2003 culture cycle. Of the 32 reservoirs stocked, reliable data on harvest were obtained from 25 reservoirs. Fish yield ranged from 53 to 1801 kg ha⁻¹ and the yields of non-perennial reservoirs in southern region were significantly (P < 0.05) higher than those in the northern region. Naturally-recruited snakehead species contributed the catches in northern reservoirs. Fish yield was curvilinearly related to reservoir area (P < 0.05), and a negative second order relationship was evident between SD and yield (P < 0.05). Chlorophyll-a and fish yield exhibited a positive second order relationship (P < 0.01). Bighead carp yield impacted positively on the total yield (P < 0.05), whereas snakehead yield impact was negative. Bighead carp, common carp and rohu appear suitable for poly-culture in non-perennial reservoirs. GIFT strain O. niloticus had the lowest specific growth rate among stocked species and freshwater prawn had a low return.

Culture-based fisheries in non-perennial reservoirs of Sri Lanka: influence of reservoir morphometry and stocking density on yield

Culture-based fish yield in non-perennial reservoirs of Sri Lanka was related to reservoir morphometry and stocking density. The reservoirs were stocked mainly with fingerlings of one Chinese and three Indian major carp species, common carp, Cyprinus carpio L., and the genetically improved farmed tilapia strain of Nile tilapia, Oreochromis niloticus (L.), at four pre-determined species combinations and a range of stocking densities [SD (fingerlings ha⁻¹)]. Twenty-three reservoirs were harvested successfully at the end of the culture period of 2002–2003. Basic limnological and morphometric parameters, including shoreline development (D_L) and shoreline area ratio (R_LA), were estimated for each of the 23 reservoirs. Bray–Curtis similarity and non-metric multidimensional scaling using mean values of limnological data revealed that reservoirs could be ordinated into two major clusters, one with intact sample distribution due to similar trophic characteristics and the other with scattered sample distribution. Reservoirs in the cluster with similar trophic characteristics showed significant correlation (P < 0.05) between R_LA and total fish yield (Y). A multiple regression equation, Y = −693 + 4810 R_LA + 0.484 SD, was generated to estimate fish harvest in relation to SD.

A genomic approach to understanding the cause and effect of annual ryegrass toxicity

Annual Ryegrass Toxicity (ARGT) is a potentially lethal disease affecting livestock grazing on pastures or consuming fodder that include annual ryegrass (Lolium rigidum) contaminated with corynetoxins. The corynetoxins (CTs), among the most lethal toxins produced in nature, are produced by the bacterium Rathayibacter toxicus that uses a nematode vector to attach to and infect the seedheads of L.rigidum. There is little known of the factors that control toxin production. Several studies have speculated that a bacteriophage specific to R.toxicus may be implicated in CT production. We have developed a PCR-based assay to test for both bacterium and phage in ryegrass material and results indicate that there is a correlation between phage and bacterial presence in all toxic ryegrass samples tested so far. This PCR-based technique may ultimately allow for a rapid, high-throughput screening assay to identify potentially toxic pastures and feed in the field. Currently, ~80% of the 45 Kb genome has been sequenced an investigation to further elucidate its potential role in toxin production.Furthermore, specific alterations in gene expression as a result of exposure to CTs or the closely related tunicamycins (TMs), which are commercially available and considered biologically indistinguishable from CTs, will be evaluated for use as biomarkers of exposure. The effects of both toxins will be analysed in vitro using a rat hepatocyte cell line and screened on a low-density DNA micro array “CT-Chip” that contains <100 selected rat hepatic genes. The results are expected to further define the bioequivalence of CTs and TMs and to identify levels of exposure that are related to specific toxic effects or have no adverse effect on livestock.