Mass propagation of Satyrium nepalense D.Don.—A medicinal orchid via seed culture

An in vitro plant regeneration protocol was successfully established in Satyrium nepalense, a terrestrial orchid by culturing immature seeds from unripe fruits. Seeds were germinated on Murashige and Skoog, Knudson C and Knudson C modified Morel medium. The germination of the seeds and development of protocorm was highest in MS medium (86.7%) followed by Knudson C modified Morel medium (74%) and Knudson C medium (61.2%). Among the cytokinins tried for multiple shoot induction from the protocorm, 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ) was found to be superior. Indole-3-butyric acid (IBA) was effective for inducing healthy roots. Well-developed plantlets were hardened in vermicompost (leaf litter + cow dung 1:1), sand and coconut husk (1:1:1).     

A SCAR marker linked to the N gene for resistance to root knot nematodes (Meloidogyne spp.) in pepper (Capsicum annuum L.)

The root-knot nematodes (Meloidogyne spp.) are important nematode pests and cause serious diseases in pepper in the world. No molecular markers linked to the nematodes resistance N gene have been reported. In this paper, ‘Carolina Wonder’ (Capsicum annuum L.), a sweet pepper line resistant to root-knot nematode with N gene, ‘20080-5-29’ (C. annuum L.), an inbred line susceptible to root-knot nematode with good horticultural characteristics, and their F₂ progeny with 320 individuals were used as materials. Evaluation of resistance and susceptibility of parental lines, F₁ and F₂ progeny inoculated with root-knot nematodes (Meloidogyne incognita) were carried out. ‘Bulked segregant analysis’ method was used to search for polymorphic markers from 512 pairs of AFLP primers. Based on the assessment of resistance and susceptibility and polymorphism of the AFLP marker in F₂ population, the genetic linkage distance between the AFLP marker and the N gene was estimated. One AFLP marker E39/M41-339 was obtained and transferred to a SCAR marker amplifying a 315 bp DNA fragment linked to the N resistant allele and a 331 bp fragment linked to the N⁺ susceptible allele. The distance between the molecular marker and the nematodes resistance N gene is 6.3 cM. This research delivered a valuable tool for the marker assisted selection of nematodes resistance in pepper.     

Condition assessment of lamprey populations in the Yorkshire Ouse catchment,north‐east England,and the potential influence of physical migration barriers

• 1. River lamprey (Lampetra fluviatilis) and sea lamprey (Petromyzon marinus) are designated features of the River Derwent Special Area of Conservation (SAC) and the Humber Estuary (a possible SAC). This study determined the condition of lamprey populations in the Yorkshire Ouse catchment by assessing the species composition, distribution, abundance and size‐structure of larval (ammocoete) populations in five major tributary rivers (Derwent, Swale, Ure, Nidd and Wharfe).
• 2. According to EU condition assessment criteria, Lampetra populations (assumed to be mostly river lamprey) are at present in favourable condition, with site mean (±SE) densities ranging from 2.7 (±1.2) to 160.3 (±50.5) individuals m^?2 (all river means >2 individuals m^?2), and at least two size (≈age) classes present in optimal microhabitats. By contrast, no sea lamprey larvae were recorded, suggesting that populations of this species are in unfavourable condition.
• 3. Actions to protect and enhance nationally or internationally important stocks must be implemented from at least a catchment perspective, because many of the issues affecting such species are not localized. With respect to lampreys, particular attention should be given to protecting spawning and nursery habitats, improving water quality, reducing impingement at abstraction points, preventing exploitation at spawning grounds and increasing passage at potential physical obstructions.

Copyright © 2007 John Wiley & Sons, Ltd.     

Effects of dietary caffeine on growth,body composition,somatic indexes,and cerebral distribution of acetyl‐cholinesterase and nitric oxide synthase in gilthead sea bream (Sparus aurata), reared in winter temperature

This study aims at examining the effect of caffeine administration on growth, feed efficiency, and consumption of sea bream (Sparus aurata), reared in winter temperatures. Moreover, it is questioned whether caffeine has a central action in the brain and its effects are partly mediated via central brain mechanisms. For this, we studied the influences of caffeine treatment on the cerebral pattern of the cholinergic neurotransmission and the novel neuromodulator nitric oxide (NO), by means of acetyl‐cholinesterase (AchE) and nitric oxide synthase (NOS) histochemistry. Five different diets containing 0.0, 0.1, 1.0, 2.0 and 5.0 g caffeine kg^?1 of diet were administrated to five groups of fish. Caffeine adversely affected sea‐bream growth at a concentration higher than 1 g kg^?1 diet and increased feed conversion ratio in the treatments of 2 and 5 g kg^?1 (P < 0.05). The daily consumption of feeds was similar to all groups, indicating that caffeine did not influence diet palatability. AChE‐ and NADPH‐diaphorase histochemistry showed densely labeled cells and fibers mainly in dorsal telencephalon, preoptic, pretectal, hypothalamic areas, optic tectum, reticular formation, cerebellum and motor nuclei. When compared with matched caffeine‐treated animals, no differences in the histochemical pattern and cell densities of cerebral AChE and NADPH‐diaphorase were found.     

Application of sperm cryopreservation in selective breeding of the Pacific oyster,Crassostrea gigas (Thunberg)

The robustness of Pacific oyster, Crassostrea gigas (Thunberg), sperm cryopreservation in the context of selective breeding based on family lines was investigated. Irrespective of egg density, high fertilization success was achieved with cryopreserved sperm when sperm:egg ratios of 1000:1 to 10 000:1 were used. Variation among replicate runs on the same oyster batches was minimal, indicating that cryopreservation and larval rearing procedures were repeatable. Twenty independent single male–female crosses were made to assess the utility of cryopreserved sperm in selective breeding. The fertility of unfrozen sperm was generally a poor predictor of cryopreserved sperm fertility. Based on D‐larval yields, 17 of the 20 crosses were likely to yield adequate spat for selective breeding (>10⁵ D‐larvae from 1 million eggs), two were marginal (5 × 10⁴ D‐larvae) and one was inadequate (4 × 10³ D‐larvae). An alternative fertilization strategy to improve D‐yield from a given number of sperm was then tested. Fertilizing 10 million eggs at a sperm:egg ratio of 200:1 increased the total D‐yield when compared with fertilizing 1 million eggs at a sperm:egg ratio of 2000:1 for the same male–female pair. We conclude that, despite wide variation in fertility, cryopreserved sperm is useful for family production.     

Estimating reference evapotranspiration with atmometers in a semiarid environment

Reference evapotranspiration (ET₀) estimations require accurate measurements of meteorological variables (solar radiation, air temperature, wind speed, and relative humidity) which are not available in many countries of the world. Alternative approaches are the use of Class A pan evaporimeters and atmometers, which have several advantages compared to meteorological stations: they are simple, inexpensive and provide a visual interpretation of ET₀. The objectives of the study were to compare the evaporation from atmometers (ET_gage) with the evapotranspiration estimated by the FAO-56 Penman-Monteith equation (ET_0PM) and to evaluate the variability between three modified atmometers of a commercial model. Comparison between daily ET_gage measured by the atmometer and ET_0PM showed a good correlation. However, ET_gage underestimated ET_0PM by approximately 9%. Differences between ET_gage and ET_0PM ranged from −2.4 to 2.2 mm d⁻¹ while the mean bias error was −0.41 mm d⁻¹. Underestimations occurred more frequently on days with low maximum temperatures and high wind speeds. On the contrary, atmometer overestimations occurred on days with high maximum temperatures and low wind speeds. Estimates of ET₀ using the atmometer appeared to be more accurate under non-windy conditions and moderate temperatures as well as under windy conditions and high temperatures. Atmometers 2 and 3 overestimated the evaporated water by atmometer 1 with a maximum variability of cumulative water losses of 4.5%. A temperature-based calibration was performed to improve the atmometer accuracy, using maximum temperature as an independent variable, with good results.     

Irrigation management practices of cabbage farmers in Botswana using saline groundwater

All horticultural production in Botswana is irrigated, as average rainfall is less than average reference evapotranspiration in all parts of the country for all of the year, and most of the irrigation in the country is used for horticultural crops. In the main, the water used for irrigation comes from groundwater, and much of this can be classified as moderately saline. A survey was carried out of 60 cabbage farmers in five regions of the country to identify the level of salinity of irrigation water and the irrigation practices used. It was concluded that there was no relationship between the irrigation rate (average irrigation depth divided by average irrigation interval) and the salinity of the water used or the salinity of the soil, such that some farmers were over-irrigating and others under-irrigating. This means that water is being wasted and yields are likely to be reduced. In addition, there is a risk of long-term build-up of salinity and soil degradation.     

Mineral content of three olive cultivars irrigated with treated industrial wastewater

The reuse of saline treated industrial wastewater generated by textile firms mixed with municipal domestic effluent for irrigation was used to asses its effect on the mineral content of three olive (Olea europaea L.) cultivars under greenhouse and field conditions during two complete vegetative cycles. Chemical analysis of the treated wastewater indicated that the element concentrations fall within the permissible range of irrigation water used for plants. However, little impermissible accumulation of Na and Mg higher than the recommended maximum concentration was observed. Irrigation water with six electrical conductivities (EC = 0.78, 1.0, 2.0, 3.0, 4.0 and 5.0 dS m⁻¹ in treatments T₀, T₁, T₂, T₃, T₄, T₅, respectively) were compared in the greenhouse experiment. The olive trees in the field experiment were trickle irrigated with potable water and treated wastewater (average EC = 4.2 dS m⁻¹). The results of the greenhouse experiment showed that leaf N, Cu, Mn, Fe, Pb, and Na contents increased with increasing salinity of the treated wastewater. This increase was accompanied with a decrease in K and Mg contents. Leaf Ca and Cl concentrations were not considerably affected. Ion analysis in roots indicated that the contents of P, Na, Cl, Mn, and Pb increased while K decreased as treated wastewater salinity increased. Consequently, in most cases T₄ and T₅ gave a highly significant increase or decrease in accumulation of the previously mentioned minerals. A considerable variation in the studied cultivars was noticed. ‘Nabali’ was considered the most tolerant cultivar for the high salinity levels of the treated wastewater; its transporting selectivity of Na from root to leaf was higher and more Na was retained in the roots. Tissue analysis of leaves indicated that the element concentrations were within the adequate levels except those of Fe in ‘Nabali’ and ‘Manzanillo’, Na in ‘Improved Nabali’ and Cu in ‘Nabali’ and ‘Manzanillo’. In view of these findings, the negligible accumulation of minerals in leaves and roots indicated that this kind of textile effluent can be used as a valid alternative for irrigation of olive orchards with continuous monitoring of mineral levels.     

Changes in epidemiological patterns of sea lice infestation on farmed Atlantic salmon, Salmo salar L., in Scotland between 1996 and 2006

Analyses of a unique database containing sea lice records over an 11 year period provide evidence of changing infestation patterns in Scotland. The data, collected from more than 50 commercial Atlantic salmon farms, indicate that both species of sea lice commonly found in Scotland, Lepeophtheirus salmonis and Caligus elongatus, have declined on farms over the past decade. Reductions for both species have been particularly marked since 2001 when more effective veterinary medicines became available. Treatment data were also available in the database and these show a growing trend towards the use of the in-feed medication emamectin benzoate (Slice), particularly in the first year of the salmon production cycle. However, this trend towards single product use has not been sustained in 2006, the latest year for which data are available. There is some evidence of region to region variation within Scotland with the Western Isles experiencing higher levels of infestation. However, compared to the levels observed between 1996 and 2000, all regions have benefited from reduced lice infestation, with the overall pattern showing a particular reduction in the second and third quarters of the second year of production.