A filter-assisted culture method for isolation of Treponema spp. from bovine digital dermatitis and their identification by MALDI-TOF MS

Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. (T. pedis, T. phagedenis, and T. medium) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS–based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.     

Comparing mating designs to restore seed production of interspecific hybrids between Trifolium repens (white clover) and Trifolium uniflorum

Interspecific hybrids between Trifolium uniflorum and cultivated white clover (Trifolium repens) have highly useful characteristics for temperate pastoral systems derived from both parent species. However, the early hybrids (F₁ and BC₁) also have unacceptably poor seed production for commerce. This study analysed the basis for the poor seed production and investigated breeding strategies for overcoming the problem. The BC₁F₁ generation produced lower‐than‐expected numbers of heads per plant and seeds per floret. Backcrossing of selected hybrids to white clover corrected these deficiencies and created new variation. Seed numbers were also returned to near target levels by recurrent selection within the BC₁ generation. Thus, it was possible to retain a theoretical average of 25% of T. uniflorum genome and still achieve high seed production per plant. The BC₁F₂ and BC₂F₁ generations produced high seed numbers per plant, along with reasonable variation. Both of these second‐generation hybrid forms have high reproductive potential and should be the focus for the selection of the desired combinations of agronomic and seed production traits.     

Vacuum Drying for Extending Litchi Shelf-Life: Vitamin C,Total Phenolics,Texture and Shelf-Life Assessment

In an attempt to obtain shelf-stable litchi fruit with preserved nutritional quality and good sensory features, quarters of peeled and pitted fruits were vacuum dried at 50, 60 and 70 °C at a constant pressure of 8.0 kPa. The product was assessed for its vitamin C, total phenolics and texture (hardness). In addition, the product with the best texture was assessed for its shelf-life by means of accelerated testing. Results suggest that vacuum dried litchi retained almost 70% of the vitamin C and total phenolics when compared to frozen fruits (control). Vitamin C and phenolic compounds content significantly decreased with drying, while no difference was found between different drying temperatures. Hardness increased with drying temperature. The sample dried at 70 °C presented crispness, which is a desired quality feature in dried fruit products. This sample was subjected to shelf-life evaluation, whose result suggests a shelf-life of eight months at 23 °C. Total color change (CIE ΔE₀₀) was the expiry criterion. Vacuum drying was a suitable technique for producing shelf-stable litchi fruit with good texture while preserving its desirable original nutrients. Consumption of vacuum dried litchi may be beneficial to health due to its remarkable content of phenolic compounds and vitamin C.     

Anthocyanin and flavonoid production from Perilla frutescens: pilot plant scale processing including cross-flow microfiltration and reverse osmosis

Extraction and concentration at a pilot plant scale of anthocyanins and flavonoids from Perilla frutescens var. frutescens harvested in the Guangzhou area of China were investigated. The study of extraction efficiency using mineral acids and organic acids showed that 0.01 mol/L nitric acid was the most suitable to extract flavonoids from this slightly red leaf cultivar. The red extract contained 12 mg/L (as cyanidin equivalent) anthocyanins and other flavones. The multistep process included cross-flow microfiltration (CFM) with a ceramic type membrane, reverse osmosis (RO), and rotating evaporation (RE). The filtration fluxes were high and constant for CFM (150 L/h/m2 at 0.6 b) and for RO (22 L/h/m2 at 40 b). The red extract was concentrated 9.4 times by RO and then 5.4 times by RE. It contained 422 mg/L anthocyanins, representing 77% of the total extracted anthocyanin. The proportion of flavonoids was found unchanged during processing. The concentrated extract showed a pH of 2.7, and its free acidity was found to be 46% of the acidity added for extraction, because of the buffering capacity of the extract. At the concentration level reached, a crystallized deposit occurred and was identified as tartrate.     

Changes in fatty acid and sterol composition during oogenesis in the pearl oyster Pinctada margaritifera

The fatty acid and sterol composition of the oyster Pinctada margaritifera during oogenesis and in eggs was analysed. No major differences were observed during oogenesis, but the egg composition was significantly different from that of gonads. The amount of saturated fatty acids was the highest in eggs and the C16:0 predominant (P<5%); by contrast, the amount of 22:6(n‐3) was significantly lower (P<5%) than in gonads. No major differences were observed for the polar lipid (PL) composition during oogenesis. The main free sterols in gonads and eggs were cholesterol and brassicasterol. Among free sterols, the proportion of cholesterol diminished continuously from the beginning to the end of gonad maturation, and this decrease persisted in eggs after spawning. Cholesterol represented 40% to 55% of the sterol ester encountered in gonad and eggs. This study allowed us to investigate the fatty acid and sterol composition during oogenesis of the pearl oyster P. margaritifera, leading to a clearer understanding of the nutritional requirements of pearl oyster during the reproduction process.     

Complete Larval Development of the Fresh Water Prawn Macrobrachium vollenhovenii in Cameroon

The African River prawn Macrobrachium vollenhovenii is the largest of the local Palaemonidae prawns along the Cameroonian coast and an important target species for fisheries and aquaculture. A preliminary study on larviculture of this species was carried out at the IRAD-AQUASOL hatchery in Kribi, Cameroon. Berried females M. vollenhovenii were obtained from the Lobe and Lokoundje Rivers in the Southern Region, typical Lower Guinea rainforest streams. The larval rearing system consisted of a single recirculation system of four 65 L cylindrical plastic containers connected to a submerged biological filter container (65 L). Experiments were conducted three times in the same condition, with two batches of larvae reared separately in duplicate per experiment, at a density of 40/L, temperature 26°C, and salinity 16%. Larvae were fed from 1 day after hatching until metamorphosis to postlarvae (PL) with newly hatched Artemia nauplii, three times daily at the rate of 5 nauplii per mL water. The number of newly hatched larvae per female body weight unit was not affected by the female size and ranged between 531 to 1349 larvae g^?1. The six batches succeeded from hatching to metamorphosis, and the 11 distinct larval stages described for M. rosenbergii were found. Larvae in all the batches developed more or less at the same pace up to stage V. However, a clear difference in the timing of appearance of the developmental stages was observed between batches and individuals of the same batch during the subsequent stages. The transition from stage V to stage VI and from stage IX to stage X was longer than the passage to other stages and seems to be the critical rearing period of M. vollenhovenii. From day 20 onward to sampling, the larval stage index (LSI) showed that larval development of the batch 1 was significantly faster than for all other batches. The time for first appearance of postlarvae was variable between batches (41–74 days), and batch 1 larvae passed through metamorphosis within a shorter span of time (41 days). Survival up to postlarvae was variable among batches, ranging between 3%–9%, and was better in batch 1 (9.31 ± 1.09). The general poor performance in terms of survival and metamorphosis rates of larvae seems to be related to the wild condition of broodstock. To improve performance of larvae, domestication of stock may be necessary. This result is the first recorded success in larviculture of M. vollenhovenii in Cameroon.     

From shallow to deep waters: habitats used by larval lampreys (genus Petromyzon and Lampetra) over a western European basin

Abstract –  Habitat requirements of lamprey ammocoetes (Petromyzon marinus and Lampetra genus) were investigated, for the first time, from shallow to deep waters, at different spatial scales across the Gironde‐Dordogne continuum, thanks to a water suction dredge. Fish‐habitat relationships were assessed through two complementary statistical analyses: habitat‐use curves and habitat suitability models using the Boosted Regression Trees (BRT) technique. Analyses were performed on a small‐size data set that was characterised by the low prevalence of lamprey. The sea lamprey larvae occurred in deeper areas than their Lampetra genus counterparts. ‘Pools’ of 2 m’ depth and more were optimal habitats for the former species. Among the environmental variables retained to model lamprey occurrences, the mesohabitat (a categorical variable) was demonstrated to be highly influential, in terms of fine grain‐size substratum and vegetation cover. These preliminary results suggest that monitoring using the water suction dredge method may contribute to sea lamprey conservation.