Bovine cysticercosis-Development of a real-time PCR to enhance classification of suspect cysts identified at meat inspection

Laboratory confirmation methods are important in bovine cysticerosis diagnosis as other pathologies can result in morphologically similar lesions resulting in false identifications. We developed a probe-based real-time PCR assay to identify Taenia saginata in suspect cysts encountered at meat inspection and compared its use with the traditional method of identification, histology, as well as a published nested PCR. The assay simultaneously detects T. saginata DNA and a bovine internal control using the cytochrome c oxidase subunit 1 gene of each species and shows specificity against parasites causing lesions morphologically similar to those of T. saginata. The assay was sufficiently sensitive to detect 1 fg (Ct 35.09 +/- 0.95) of target DNA using serially-diluted plasmid DNA in reactions spiked with bovine DNA as well as in all viable and caseated positive control cysts. A loss in PCR sensitivity was observed with increasing cyst degeneration as seen in other molecular methods. In comparison to histology, the assay offered greater sensitivity and accuracy with 10/19 (53%) T. saginata positives detected by real-time PCR and none by histology. When the results were compared with the reference PCR, the assay was less sensitive but offered advantages of faster turnaround times and reduced contamination risk. Estimates of the assay's repeatability and reproducibility showed the assay is highly reliable with reliability coefficients greater than 0.94. Crown Copyright (C) 2013 Published by Elsevier B.V. All rights reserved.

Modern dinoflagellate cysts found in surface sediments of Santa Cruz Island, Galapagos

Modern dinoflagellate cysts in surface sediments collected on Santa Cruz Island, Galapagos, are described, along with other palynomorphs such as microforaminiferal linings, tintinnid loricae, copepod eggs and acritarchs including Domasiella-like micro-remains and Halodinium spp. The dinoflagellate cyst assemblages mainly consisted of Spiniferites cf. scabratus (gonyaulacoid) followed by Brigantedinium spp. and Selenopemphix quanta (peridinioids). No gymnodinioid cysts were found. No remarkable differences in cyst composition and densities were recognized between stations. The cyst assemblages were characterized by low species diversity and low cyst concentrations in comparison with the Pacific coast of Guatemala and Peru. CDF Contribution Number 1019.

Successful inoculation of Artemia and production of cysts in the coastal saltpans of Bangladesh II

Bangladesh has no naturally occurring Artemia, and all the growing shrimp hatcheries of the country depend entirely on import of cysts from foreign countries. Following successful inoculation of Artemia and production of cysts for the first time in this country in a coastal saltpan (at Chanua, Banskhali) by the senior author (in 1989-90), a similar second attempt was made under this programme in a saltpan (1000 m super(2)) of Demoshia, Chakaria, Cox's Bazar, Bangladesh between January and April 1992. A total of 1639.9 g (dry weight) of cysts (i.e. 5.46 kg DW/ha/month) have been produced using the Red Jungle Brand, whereas the previous attempt obtained 517 g of cysts (i.e. 2.07 kg DW/ha/month) using the Great Salt Lake Brand.

Effect of decapsulation on viability and hatching performance of Artemia cysts at different salinity levels

Artemia cysts were produced from the traditional solar salt works of Bangladesh through different fertilization treatments were tested for viability and hatching performance in different forms, such as processed and preserved, processed and decapsulated and unprocessed and undecapsulated. Decapsulated cysts performed maximum hatching (86.0%) in 20ppt salinity during 48 hours of incubation. The hatching percentage by the unprocessed and undecapsulated cysts were very low (12.0- 18.7%) in all the tested salinity grades.

Effects of culture treatments on HUFA levels in Artemia cysts

Artemia cysts (of GSL, Utah, USA origin) were produced from the modified traditional solar salt works of Bangladesh during winter months through different feeding/fertilization treatments (T1, T2, T3, T4 ) were analyzed to understand the effects of treatments on their fatty acid profile. Palmitic, Linolenic, Eicosapantaenonic and Docohexaenoic acids (mg/g. DW) were found highest for the cysts in T1 (16.0% ±1 .36), T2 (14.7% ±0.47), T3 (4.7% ±0.40) and T4 (0.7% ±0.06) treatments, respectively. High amount of 18:3(n-3) acids in the cysts of all sources proves to be freshwater type of the cysts. The presence of marine type essential fatty acids in the cysts of all sources were found low for 20:5n-3 (3.7-4.7%) and very low for 22:6n-3 (0.09-0.7%). No significant variation was observed for 16:0 acids within the treatments, but for 18:3(n-3) acid, the variation was found highly significant (P= 0.0052) between T2 and T4 treatments. For 20:5(n-3), only variation between T2 and T4 was found insignificant (P=0.1161), but between other treatments, significant variation was observed between T2 and T4 (P=0.0241), T2 and T4 (P=0.0022) and T1 andT4 (P=0.0161). No significant variation was found in other treatments.

Successful inoculation of Artemia and production of cysts in man-made salterns in the Philippines

The authors report on inoculation experiments of Artemia nauplii and young adults of the San Francisco Bay strains in earthen fish ponds. The test inoculated proved successful where water salinity ranges from 20 to 32 o/oo during the start of the rainy season in the Philippines.

The synthesis of astaxanthin esters, independent of the formation of cysts, highly correlates with the synthesis of fatty acids in Haematococcus pluvialis

The compositions and contents of astaxanthin esters and fatty acids in four types of Haematococcus pluvialis cells were studied by HPLC and GC-MS. Results showed that the synthesis and accumulation of astaxanthin was independent of the formation of cysts, but was highly correlated with the synthesis and accumulation of fatty acids, though it is an well known phenomenon that the accumulation of astaxanthin is usually accompanied by the formation of cyst. The red cysts contain more than 30% of fatty acids, with 81% of the unsaturated fatty acids. Taken together, besides a resource of astaxanthin, H. pluvialis would be a good resource of valuable fatty acids.

Dinoflagellate Cysts as Indicators of Cultural Eutrophication in the Oslofjord, Norway

Dinoflagellate cyst records were analysed from four sediment cores from the inner Oslofjord. The cores covered the pre-industrial period, and the most important period of human population growth associated with industrial development of the region, from the mid-1800s to the present, including the reported development of cultural eutrophication. Comparisons between the cyst records and the known history of eutrophication suggest cyst signals that should prove useful for tracing the development of eutrophication. The eutrophication signal consisted of a doubling of total cyst concentration, and a marked increase in one species in particular,Lingulodinium machaerophorum(from <5 to around 50% of the assemblages) with increased eutrophication. In the core considered most representative of general water quality in the inner fjord, these trends reversed back to pre-industrial levels during the 1980s and 1990s when improved sewage treatment took effect.

Dinoflagellate cysts as potential indicators of industrial pollution in a Norwegian Fjord

Variation in dinoflagellate cyst assemblages through the last approximately 300 years was studied in two sediment cores, one from the heavily polluted Frierfjord, and one from the adjoining, relatively unpolluted Brevikfjord, in order to docu1ent possible dinoflagellate responses to pollution. Changes in the cyst-flora were compared with historical information on the development of industry and also with geochemistry of the sediments, reflecting aspects of pollution. In the Frierfjord core, increasing pollution was accompanied by a decrease in cyst concentration, possibly reflecting reduced production, at least of dinoflagellates, and a shift toward more heterotrophic species, possibly reflecting reduced light penetration in the euphotic zone, or increased production of prey for the heterotrophs. These trends seem to have reversed as pollution decreased after about 1975, suggesting that cyst assemblages contain signals that may prove useful for tracing the development of pollution. Cyst assemblages in the Brevikfjord core only showed minor changes.

Phylogenetic position and morphology of thecae and cysts of Scrippsiella (Dinophyceae) species in the East China Sea

Resting cysts of the marine phytoplanktonic dinoflagellate Scrippsiella spp. are encountered in coastal habitats and shallow seas all over the world. Identification of Scrippsiella species requires information on cyst morphology because the plate pattern of the flagellated cell is conserved. Cysts from sediments of the East China Sea were identified based on traits from both the cysts and the thecal patterns of germinated cells. Calcareous cysts belonged predominantly to S. trochoidea (F. Stein) A. R. Loebl., S. rotunda J. Lewis, and S. precaria Montresor et Zingone. The former two species also produced smooth and noncalcified cysts in the field. A new species, S. donghaienis H. Gu sp. nov, was obtained from six noncalcified cysts with organic spines. These cysts are spherical, full of pale white and greenish granules with a mesoepicystal archeopyle. The vegetative cells consist of a conical epitheca and a round hypotheca with a plate formula of po, x, 4', 3a, 7 '', 6c (5c + t), 6 s, 5''', 2'''' and are morphologically indistinguishable from S. trochoidea. Results of internal transcribed spacer (ITS) sequence comparisons revealed that S. donghaienis was distinct from the S. trochoidea complex and appeared nested within the Calciodinellum/Calcigonellum clade. Culture experiments showed that the presence of a red body in the cyst and the shape of the archeopyle were constant within cell lines from one generation to the next, while the morphological features of the cyst wall, such as calcification and spine shape, appeared to be phenotypically plastic.