Importance of experimental design in detecting and measuring stresses in marine populations

Environmental sampling to detect and measure the sizesof stresses is made extremely difficult by naturalspatial and temporal variations in most possibleecological measurements. Sound application ofprinciples of experimental design is therefore acrucial component of any study. Temporal replicationbefore and after a disturbance is necessary to ensurethat accurate estimates are made of averageconditions. Otherwise, sampling will only reveal whathappens at the particular time sampled. Spatialreplication of reference or control locations isessential to prevent confounding any interpretation ofa difference between a disturbed and a controllocation. Unless locations are replicated, anydifference may be a stress or may be caused by anyother processes causing a difference. If there is nodifference, this does not mean there is no stress. Environmental stresses are manifested as statisticalinteractions due to changes in time being different indisturbed as opposed to control locations (or,equally, spatial differences between the disturbed andcontrol locations being different before and after thedisturbance). Features of experimental design thatallow the most reliable interpretation of suchinteractions should be the focus of planning ofstudies. Here, an ideal design is considered, withdiscussion of what is lost when (as often happens),idealism cannot be maintained. The discussionidentifies that sound planning, good design andadherence to precautionary principles all require thatunambiguous hypotheses are identified in advance ofsampling. In contrast to other approaches to analysisof stress, this approach has an explicit logicalstructure, assists to prevent confounding ininterpretations, identifies the sizes of stress thatwould elicit different responses from decision-makersand allows advance calculation of power to detect theanticipated stress for any chosen size of sampling. As a result, it is clear that more emphasis is neededon advance categorization of the types, directionsand, above all, magnitudes of stress that might matterin any particular case. Explicit statements about thesize of a possible stress will allow better planning,clearer guidance for development of programmes ofsampling and their interpretation and would reduce theuncertainty that categorizes much of environmentalanalysis.     

Glutathione metabolism at the blood-cerebrospinal fluid barrier

Glutathione metabolism and transport in the choroid plexus were probed by determining the effects of administration to rats of several compounds (buthionine sulfoximine, L-2-oxothiazolidine-4-carboxylate, L-(alpha 5,5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazole acetic acid, gamma-glutamyl alanine, and glutathione monoethyl ester) on the levels of glutathione and cysteine in the cerebrospinal fluid. The findings indicate that glutathione is actively metabolized in the choroid plexus by pathways similar to those in kidney and other tissues. The level of glutathione in the cerebrospinal fluid can be decreased or increased by giving compounds that do not, under similar conditions, appreciably alter total brain levels of glutathione. Glutathione monoethyl ester is effectively transported into the cerebrospinal fluid.     

Temporal and stoichiometric patterns of algal stimulation of litter-associated heterotrophic microbial activity

1. Periphyton communities associated with submerged plant detritus contain interacting autotrophic and heterotrophic microbes, and are sites of extracellular enzymatic activity. The strength and nature of these interactions might be expected to change over time as microbial communities develop on plant litter. Microbial interactions and enzymatic activity can be altered by nutrient availability, suggesting that litter stoichiometry could also affect these phenomena.
2. We grew wetland plants under ambient and nutrient-enriched conditions to generate plant litter of differing nutrient content. In two experiments, we investigated: (1) the influence of algal photosynthesis on fungal and bacterial production and the activities of four extracellular enzymes throughout a 54-day period of microbial colonisation and growth; and (2) the influence of litter stoichiometry on these relationships.
3. Ambient and nutrient-enriched standing-dead plant litter was collected and then submerged in wetland pools to allow for natural microbial colonisation and growth. Litter samples were periodically retrieved and transported to the laboratory for experiments manipulating photosynthesis using the photosystem II inhibitor DCMU (which effectively prevents algal photosynthetic activity). Algal (¹⁴C-bicarbonate), bacterial (³H-leucine), and fungal (¹⁴C-acetate) production, and β-glucosidase, β-xylosidase, leucine aminopeptidase, and phosphatase activities (MUF- or AMC-labelled fluorogenic substrates) were measured under conditions of active and inhibited algal photosynthesis.
4. Photosynthesis stimulated overall fungal and bacterial production in both experiments, although the strength of stimulation varied amongst sampling dates. Phosphatase activity was stimulated by photosynthesis during the first, but not the second, experiment. No other enzymatic responses to short-term photosynthesis manipulations were observed.
5. Microbial communities on high-nutrient litter occasionally showed increased extracellular enzyme activity, fungal growth rates, and bacterial production compared to communities on non-enriched litter, but algal and fungal production were not affected. Litter stoichiometry had no effects on fungal, bacterial, or enzymatic responses to photosynthesis, but the mean enzyme vector analysis angle (a measure of P- versus N-acquiring enzyme activity) was positively correlated to litter N:P, suggesting that elevated litter N:P led to an increase in the relative activity of P-acquiring enzymes.
6. These results supported the hypothesis that algal photosynthesis strongly influences heterotrophic microbial activity on macrophyte leaf litter, especially that of fungi, throughout microbial community development. However, the strength of this photosynthetic stimulation does not generally depend on small differences in litter nutrient content.
7. Stimulation of microbial heterotrophs by algal photosynthesis could drive diurnal shifts in periphyton community and aquatic ecosystem function, as well as linking green (photoautotroph-based) and brown (detrital-based) food webs.

     

Continuous light and physiology of arctic birds and mammals

Many investigations of pineal function have been conducted on animals experiencing artificial laboratory environments. The use of species that naturally experience extreme conditions, such as constant light or darkness, is suggested as a possible way to clarify some of the confusion that exists in the pineal literature at the present time. Arctic species are adapted to an environment in which conditions of constant light are naturally replaced with periods of alternating light. Other conditions of the arctic also vary, including temperature, wind, precipitation, available food, and inter- and intra-species interactions. Arctic species must be highly attuned to their harsh environment. Four species of arctic homeotherms have particular promise as models for the study of pineal function: Snowy owls (Nyctea scandiaca ) and short-eared owls (Asio flammeus ), in contrast with most other owls, must annually switch from being day-active to being at least partially night active. The Arctic fox (Alopex lagopus ) shows regular circannual variations in several parameters alleged to be under the influence of the pineal in other species, including reproduction, activity levels, and seasonal molts. Lemmings (Lemmus trimucronatus ) have two periods of reproductive activity per year, one at the height of the constant light season, and a less intense period that coincides with minimal light. Hormonal activity and possibly pineal activity in lemmings is also implicated in periodic, large scale population fluctuations.Presented during the Seventh International Biometeorological Congress, 17–23 August 1975, College Park, Maryland, USA.