Investigation of the Role of Androstenedione-19-oic Acid in the Presence of 19-Norandrostenedione in Intact Male Horse Plasma Using Liquid Chromatography–Tandem Mass Spectrometry

A liquid chromatography–tandem mass spectrometry method was developed to confirm the presence of androstenedione-19-oic acid in intact male equine plasma and to show the source of 19-norandrostenedione in equine plasma. Androstenedione-19-oic acid was recovered from acidified plasma by liquid–liquid extraction using methyl tert-butyl ether and separated on an Ace 5 C₈ column. A triple quadrupole mass spectrometer was used to detect the analytes in negative electrospray ionization mode. Limits of detection, quantification, and confirmation of the method were 0.1, 0.5, and 1.0 ng/mL, respectively. The linear dynamic range of quantification was 0.5–50 ng/mL. The presence of androstenedione-19-oic acid was confirmed in all plasma samples obtained from intact male horses but not those from gelded and female horses; the average concentration was 3.1 ± 1.6 ng/mL, suggesting androstenedione-19-oic acid is an endogenous compound only in intact male horse plasma samples. The conversion of androstenedione-19-oic acid to 19-norandrostenedione in equine plasma was demonstrated by spiking androstenedione-19-oic acid into blank plasma and monitoring the generation of 19-norandrostenedione and its increase in concentration during storage. Results indicated that androstenedione-19-oic acid was readily converted into 19-norandrostenedione; the higher the storage temperature, the faster the conversion. The conversion was not affected by the types of plasma samples collected from gelded and female horses or by anticoagulants used in blood collection to harvest plasma. Compared with other matrices such as water, methanol, and phosphate-buffered saline, the conversion of androstenedione-19-oic to 19-norandrostenedione in equine plasma was faster, suggesting that there is an unknown factor(s) in equine plasma that enhances the conversion.     

Different reactions of central and marginal provenances of Fagus sylvatica to experimental drought

Climate extremes are expected to increase in frequency and magnitude as a consequence of global warming, threatening the functioning, services and goods of forest ecosystems. Across Europe, the ecologically and economically important tree species Fagus sylvatica is expected to suffer particularly under such conditions. The regional introduction of provenances from drier and warmer climates is one option to adapt beech forest ecosystems to these adverse effects of climate change. Marginal populations from the drought-prone southern and north-eastern edges of the species’ distribution come into focus in search of suitable candidates for Central European deciduous forests. Here, we test three marginal provenances (Spain, Bulgaria and Poland) and three provenances from the centre of the distribution range (Germany) for their response to drought in two different soil types (sand, loam) in a full factorial common garden experiment in Landau, Germany. Drought impacted all growth parameters negatively (leaf damage +22 % (percentage points), height ?40 % and diameter increment ?41 %), and the sandy substrate exacerbated this effect. However, provenances differed in their response to drought and soil type. Evidence for a local adaptation to summer drought was detected, especially in terms of mortality rates. The Bulgarian and Spanish provenance showed a stable performance under drought conditions (BG ?27 % in diameter increment; ES ?32 %), compared to the Polish (?48 %) or the most sensitive German provenances (?57 %), yet for Bulgaria on a low level of total increment. This may indicate a trade-off between drought tolerance and growth. Therefore, a sole focus on drought-resistant marginal provenances seems to not be conducive, as they might be less adapted to other climatic factors, e.g. frost, as well. However, intermixed with local Central European provenances, these may act as functional insurance in future drought-prone forest stands.     

The effects of temperature on the physiological response to low oxygen in Atlantic sturgeon

Atlantic sturgeon (Acipenser oxyrhynchus), which are bottom dwelling and migratory fish, experience environmental hypoxia in their natural environment. Atlantic sturgeon, acclimated to either 5 or 15°C, were subjected to a 1 h severe (<10 mm Hg) hypoxia challenge in order to document their physiological responses. We measured hematological parameters, including O₂ transport (hemoglobin, hematocrit), ionic (chloride, osmolality), and metabolic (glucose, lactate) variables under normoxic conditions (~160 mm Hg), immediately following a 1 h exposure to hypoxic water, and following a further 2 h of recovery from this challenge in normoxic water. In a second experiment, we assessed the opercular beat frequency before, during, and after hypoxic exposure. Hemoglobin concentrations and hematocrit were significantly different between fish held at 5°C vs. 15°C and also significantly different between normoxia prior to hypoxia and following recovery. Plasma lactate concentrations increased following hypoxia at both temperatures, indicative of an increase in anaerobic metabolism. In contrast, a significant increase in plasma glucose concentrations in response to hypoxia only occurred at 5°C, suggesting different fuel demands under different temperatures. Changes in opercular beat frequency (OBF) were dependent on temperature. At 5°C, OBF increased upon exposure to hypoxia, but returned to pre-exposure levels within 35 min for the remainder of the experiment. During hypoxia at 15°C, OBF increased very briefly, but then rapidly (within 20 min) decreased to levels below control values. Following a return to normoxia, OBF quickly increased to control levels. Overall, these findings suggest that Atlantic sturgeons are relatively tolerant to short-term and severe hypoxic stress, and the strategies for hypoxia tolerance may be temperature dependent.     

Tank culture of yellowfin tuna, Thunnus albacares: developing a spawning population for research purposes

A land-based culture facility for research on yellowfin tuna, Thunnus albacares, was developed at the Achotines Laboratory in the Republic of Panama. Six concrete tanks, and seawater and life support systems were built to maintain a yellowfin broodstock. On average, 50% of the yellowfin caught survived capture and handling, and approximately 30% became broodstock in Tank 1 (17 m diameter, 6 m depth) or Tank 2 (8.5 m diameter, 3 m depth). Each fish was tagged with a microchip implant tag, then weighed, measured, and injected with oxytetracycline (OTC) prior to stocking. Daily rations of primarily market squid, Loligo opalescens, and Pacific thread herring, Opisthonema spp., were regulated based on the feeding activity and energy requirements of the fish. Feeding activity of the broodstock decreased when the water temperatures decreased, and the fish ate decreasing daily rations and increasing calories with increasing size. Spawning occurred in both tanks within 6–8 months of capture. Spawning first occurred in Tank 1 when 24 females ranged in size from 6 to 16 kg and 65 to 93 cm fork length (FL). Spawning was intermittent during the first 2 months and occurred near daily thereafter. Tank size appeared to affect survival rates, the types of mortalities that occurred, and the growth of the fish. Survival rates after 1 year in captivity were higher, and the fish were larger, on average, in Tank 1 than in Tank 2. Most of the mortalities in Tank 1 were the result of wall strikes, which occurred more frequently after the fish reached their highest density of 0.64 kg m⁻³ and sizes greater than 96 cm FL and 19 kg. Non-linear growth models were fitted to the initial stocking sizes and final sizes of fish that died or were removed from Tank 1 during 1996–1999. Estimated growth rates in length (11–48 cm year⁻¹) for fish between 51 and 150 cm FL decreased with increasing length. Estimated growth rates in weight ranged from 9 to 19 kg year⁻¹ for fish less than 19 kg and 20–23 kg year⁻¹ for fish greater than 19 kg. The results of this work demonstrate that the stable environment of a land-based culture facility may be the preferred system for long-term maintenance of a yellowfin broodstock.     

Quantification of Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiss (Walbaum), tissues by qPCR

A qPCR assay was developed for rapid and sensitive detection of Flavobacterium psychrophilum, the aetiological agent of bacterial cold-water disease and rainbow trout fry syndrome in salmonid fish worldwide. A set of F. psychrophilum-specific primers based on 16S rRNA gene sequences was designed and validated for specific detection and quantification of DNA isolated from representative strains of F. psychrophilum. The qPCR assay exhibited a high specificity for the 16S rRNA gene of F. psychrophilum (from 4 × 10(8) down to 11 copies per reaction) but not for other Flavobacterium species or other bacteria including fish pathogens. This qPCR-based method proved to be useful in the quantification of the F. psychrophilum titre present within organs dissected out from diseased fish. As the F. psychrophilum genome contains six copies of the 16S rRNA gene, we could infer a limit of detection corresponding to two bacteria per reaction, corresponding to 800 bacteria per fish tissue sample, and therefore 20 F. psychrophilum cells mg(-1) of tissue (for sample weighing 40 mg). The qPCR assay reported here could be a useful tool for veterinary diagnostic laboratories to monitor the F. psychrophilum infection level in fish farms.     

Visualisation of standardized life-history patterns

The life‐history patterns of fish and invertebrate species are complex. But much of this complexity can be captured in simple diagrams of coastal transects, where juveniles usually occur in larger numbers in shallow waters, while adults generally inhabit deeper, offshore waters. Iconographic representations of generalized life‐history patterns and depth profiles, with specific key life‐history parameters can capture much of these standardized patterns, including spawning areas, nursery/juvenile distributions, adult distributions and spawning migrations. Several examples presented here from a wide range of habitats and ecosystems (temperate and tropical waters, demersal, deep water, pelagic and coral reefs), including an example of different stocks of the same species, illustrate some general patterns with regard to water depth and distance from shore. The present approach should be viewed as a first step towards obtaining standardized patterns about key life‐history parameters, and will hopefully lead to incorporation into management of life‐history interconnectivity between different fishery sectors or gears. This may contribute to sustainable, ecosystem‐based approaches to management by informing policy options when faced with decisions to rationalize overcapitalized fisheries.     

Behavioral response to selected feed attractants and stimulants in Pacific white shrimp, Litopenaeus vannamei

Nine commercial feeding attractants and stimulants for Litopenaeus vannamei were evaluated by observation of behavioral responses in animals allotted in one Y-maze aquarium apparatus. In the validation phase, fishmeal–Brazilian origin (FMBO); fishmeal–Peruvian origin (FMPO); blood meal (BM), meat and bone meal (MBM), squid meal (SM), fish oil (FO) and fish solubles (FS) were evaluated. There was also a control without stimulatory raw material. The tested materials were included at 3% in neutral gelatin pellets (wet basis). In each behavioral observation, two different ingredients were offered at the same time in equal quantities, being allotted in the end of each chamber's arm. In Phase II after system validation, the following commercial attractants were tested: 80% crude protein (CP) vegetable dried biomass (VDB₈₀), 68% CP vegetable dried biomass + glutamate + betaine (VDB₆₈), complex of amino acids (alanine, valine, glycine, proline, serine, histidine, glutamic acid, tyrosine and betaine) with enzymatically digested bivalve mollusk (CAA), condensed fish soluble protein (CFSP), squid liver meal (SLM), betaine (Bet), dried fish solubles–low biogenic amines (DFS_LB), dried fish solubles–high biogenic amines (DFS_HB) and whole squid protein hydrolysate (WSPH). Attractants were used at a 3% level wet basis with neutral gelatin, without any additional ingredient source available. The best four commercial attractants from this phase (CAA, CFSP, SLM and WSPH) were compared under 0.5% and 1.0% levels. In Phase I of the study, a higher percentage of choices were observed for FMPO and FMBO. BM and FO were the least chosen ingredients. In Phase II, the worst results were observed for Bet, DFS_HB and, mainly, for VDB₈₀ and VDB₆₈. When two-by-two comparisons were performed, results suggested that CFSP and CAA were the best commercial attractants tested. In the last phase, both CFSP and CAA at 1.0% level were significantly more chosen by shrimp than CFSP (0.5%), SLM (0.5 or 1.0%) or WSPH (0.5 or 1.0%). At both 0.5% and 1.0% levels, shrimp spent a similar amount of time feeding on CFSP and CAA. However, only CAA was statistically better than SLM and WSPH together. Further work is required to better elucidate the chemical drivers of chemostimulation for L. vannamei for each one of the attractants tested.     

Challenges for Multilevel Stakeholder Engagement in Public Trust Resource Governance

The trend in wildlife management over the last two decades has been to develop locally based approaches for responsiveness to local conditions, but some state wildlife agencies are finding the amount of staff time required to service this approach prohibitive. Although local engagement strategies have been lauded as assuring that public trust obligations of state government to citizens are met, we can expect that states with a local focus as their operational level of stakeholder engagement may opt to change their approach to reflect their resource limitations. We argue for comprehensive regional level effort to understand stakeholders augmented with local engagement processes where needed to deal with special circumstances in smaller areas within a region. Such an approach can be anticipated to have implications for stakeholder engagement and human dimensions research needs, which we discuss in the context of public trust resource administration and good governance of wildlife resources.     

An unholy disease in the Holy Land: The history of anthrax between the Jordan River and the Mediterranean Sea (1909–2012)

The history of recorded cases of anthrax in human beings and animals from 1909 to 2012 in the area between the Jordan River and the Mediterranean Sea is reviewed. The disease was endemic until the middle of the 20th century, but the incidence decreased thereafter, with only sporadic cases from the 1980s onwards. Human cases have not been diagnosed in the region since 1984 and the number of episodes of animal disease has reduced to less than one per year. This decline is mostly due to the disruption of the infective cycle by improved veterinary control, including vaccination, treatment and outbreak management. A policy of reactive vaccination for 10 years of affected herds and herds grazing in their proximity has been applied. No new outbreaks have been observed in such herds after the cessation of vaccination, despite continued grazing on the same sites, so it is assumed that spore survival in such areas is shorter than 10 years. This is independent of the soil composition, which is calcareous throughout most of the relevant area. However, reemergence of anthrax, even after decades, has occurred following disturbance and heavy rainfall.