Different B cell populations mediate early and late memory during an endogenous immune response

Memory B cells formed in response to microbial antigens provide immunity to later infections; however, the inability to detect rare endogenous antigen-specific cells limits current understanding of this process. Using an antigen-based technique to enrich these cells, we found that immunization with a model protein generated B memory cells that expressed isotype-switched immunoglobulins (swIg) or retained IgM. The more numerous IgM(+) cells were longer lived than the swIg(+) cells. However, swIg(+) memory cells dominated the secondary response because of the capacity to become activated in the presence of neutralizing serum immunoglobulin. Thus, we propose that memory relies on swIg(+) cells until they disappear and serum immunoglobulin falls to a low level, in which case memory resides with durable IgM(+) reserves.     

<Emphasis Type="Italic">In silico</Emphasis> genomic subtraction guides development of highly accurate,DNA-based diagnostics for <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 3 biovar 2 and blood disease bacterium

New rapid diagnostic methods are urgently needed to discriminate the quarantine pathogen Ralstonia solanacearum (Rs) race 3 biovar 2 (R3B2) from other populations of Rs that lack the adaptation to cause bacterial wilt disease in temperate regions. We used an in silico bioinformatic approach to identify several genome sequences potentially specific to R3B2 strains. Primer sets were designed to PCR-amplify sequences in these regions, and four sets were ultimately shown to be >99% accurate for detection of R3B2 strains. On the basis of these results, several primers were designed to enable development of a loop-mediated isothermal amplification assay that was rapid, technologically simple, and essentially 100% accurate for identification of R3B2 when applied to a comprehensive collection of geographically diverse Rs strains. We fortuitously found that a sequence in one of the “R3B2-specific” regions has ~90% identity to a sequence present in strains of the blood disease bacterium (BDB), a member of the Rs species complex that infects banana. Alignments of these sequences allowed design of a second PCR primer set that proved 100% accurate for identification of BDB strains when tested on the 22 BDB strains available to us. These results demonstrate the power of in silico genomic subtraction for rapid identification of population-specific DNA sequences and for the development of simple, reliable detection methods for Rs subpopulations.     

Spatial and Seasonal Variations in Nitrogen Leaching and Acidity across Four Acid-impacted Regions of the UK

This study examines the seasonal variability in levels of nitrogen (N) leaching and acidity in four acid impacted upland regions of the UK: the South Pennines, Snowdonia, Galloway and the Mourne Mountains. All regions are acidified, with median pH values of <5.5. The South Pennines exhibit excess sulphate concentrations an order of magnitude higher than those for other regions and have the lowest pH values of all the regions. Nitrate concentrations are highest in the South Pennines (seasonal medians 20–26 μeq l^?1) and the Mourne Mountains (seasonal medians 9–26 μeq l^?1). Surface water in the Mourne Mountains is the most highly N impacted in terms of the proportional contribution of N to acidity. All N species exhibit seasonality, with greater retention by catchment soils and vegetation in August. This seasonality is most marked in Snowdonia and least marked in the South Pennines. This implies that the South Pennines have reached an advanced stage of N saturation. Despite the reductions in S deposition, xSO₄ is the dominant anion for all sites in the South Pennine and Snowdonia regions. A strong negative relationship between the contribution of NO₃ to total acidity and DON was observed for all regions except the South Pennines. It is hypothesized that catchments dominated by organic rich soils produced more DON and have an increased capacity to immobilise NO₃. Seasonal nitrogen budgets show that over 60% of the N inputs are retained within catchments, and no site retains all incoming N throughout the year.     

Ecology of otters in Northern Scotland IV. A model scheme for otter Lutra lutra L. conservation in a freshwater system in aberdeenshire

Habitats for inclusion within otter havens are illustrated from a case-history study on the middle part of the Aberdeenshire River Dee. The environment is divided into breeding and rearing areas and places for non-breeding otters. One good location for a haven incorporating all three habitats includes 12–13 km of river and two nearly lochs. In this area, part of the river bank is inaccessible, and there are secluded islands and deep woods. Havens should include tiny tributaries leading to shelter where otters are likely to have their young. An otter haven in mid-Deeside planned to incorporate all these habitats may be regarded as a model for conservation that could be followed in other similar environments elsewhere.     

Preface and acknowledgment

Preface and acknowledgment     

Molecular markers and ex situ conservation of the European elms (Ulmus spp.)

The enormous losses suffered by the European elms during recent Dutch elm disease outbreaks led to concern over the conservation of elm genetic resources, and the subsequent establishment of a series of ex situ collections. However, as ex situ collections are inevitably finite in size, some consideration needs to be given to selecting which samples to include in them. To contribute towards this process for European ex situ elm collections we have undertaken genetic studies on a Europe-wide sample of 535 individuals. A major aim has been to use genetic markers to clarify the identification of samples to ensure that the ex situ collections contain a representative spread of taxonomic diversity. This is important given the paucity of mature elms in the landscape due to Dutch elm disease. The lack of mature material (critical for identification) compounds identification problems in what was already a taxonomically difficult group. Our data (derived from random amplified polymorphic DNA and inter-simple sequence repeats) have provided a useful supplement to morphology in undertaking such sample identifications. The molecular data served to highlight mis-identified samples and led to extensive revisions of sample identities within individual countries. Our results were less useful in detecting regional intra-specific genetic structure, and do not provide sufficient information for prioritising within-species sample selections.     

Evaluation of a serological test system for the diagnosis of natural Echinococcus granulosus infection in dogs using E. granulosus protoscolex and oncosphere antigens

Serum antibody responses in feral or domesticated dogs naturally infected with Echinococcus granulosus or/and other common helminths were examined in an enzyme-linked immunosorbent assay (ELISA) using antigens prepared from E. granulosus protoscoleces or oncospheres. The ELISA using the protoscolex antigen was optimised with serums from experimental dogs monospecifically infected with E. granulosus or other helminth parasites, and helminth-free dogs. Anti-protoscolex antibody was detected in 16 of 22 (72.7%) serums from feral dogs with E. granulosus burdens ranging from 300 to 302,600 worms per dog. Seven serums from feral dogs which did not harbour E. granulosus at autopsy but which originated from an endemic hydatid region were tested using protoscolex antigen, and 1 serum gave a positive reaction. One hundred and two serums from dogs known never to have been infected with E. granulosus all gave negative reactions to protoscolex antigen. The sensitivity of the ELISA test proved to be superior to that which has been achieved by arecoline purging as a method of diagnosis for E. granulosus infection in dogs. For use of the assay in hydatid control or eradication campaigns, its sensitivity can be increased by choosing a lower absorbance discrimination value above which serums are regarded as having positive reactions. However, this does introduce positive reactions of some serums from dogs infected with helminths other than E. granulosus. In further development of the assay, use of defined recombinant antigens may improve both sensitivity and specificity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Physiological responses in foals weaned by abrupt or gradual methods

Physiological responses to weaning procedures were studied in 21 foals assigned to one of five treatments: (1) abrupt, total separation of mare and foal, no preweaning creep feed (TSNC); abrupt, total separation with preweaning creep feed (TSC); partial separation of mare and foal allowing fenceline contact, no preweaning creep feed (PSNC); (4) partial separation with creep feed (PSC); and (5) control (CON) no separation of mare and foal, foals creep fed. Changes in adrenal response to exogenous ACTH, basal and peak plasma cortisol concentrations, plasma triiodothyronine (T3) and thyroxine (T4) concentrations, weight gains and feed consumption were measured. Foals on the total separation treatments had higher adrenal responses (P<.05) and pre-ACTH basal (P<.05) and post-ACTH peak plasma cortisol concentrations (P<.05) than foals on other treatments indicating they were stressed at weaning. The PSNC, PSC and CON treatments did not differ (P>.05) in any cortisol response. No treatment differences were found in thyroid hormone concentrations in this study. On partial separation treatments, creep-fed and non-creep-fed foals consumed similar amounts of feed during the first week postweaning. On total separation treatments, non-creep-fed foals consumed more feed (P<.05) than creep-fed foals. All foals without creep feed gained more weight immediately after weaning (0–2 weeks) than creep-fed foals (P<.05), reflecting higher feed intakes and possible compensatory gains. Total postweaning weight gains (0–8 weeks) of foals were not significantly affected by treatment.     

Feather picking and self-mutilation in psittacine birds.

Feather-picking and self-mutilation behaviors are common in psittacine birds. These behaviors are best defined as stereotypic behaviors or obsessive compulsive disorders. There is likely a genetic predisposition for these behaviors as reflected in the overrepresentation of a number of species of psittacines. Stereotypies most often result from a high level of arousal and the inability to respond with the appropriate natural behaviors. More specifically, conflict may be induced by the absence of releasing stimuli or target objects. On a chemical level, stereotypic behavior appears to be related to an increased dopaminergic activity and an increased dopamine turnover. Feather-picking and self-mutilation patients are challenging to treat and are most likely to respond to treatment at an early stage of development. The best results are obtained by identifying and removing the cause of conflict, enhancing the environment, using appropriate drugs when indicated, and employing counter-conditioning.     

Effect of levamisole on the clinical and immunologic responses to oral vaccine of Treponema hyodysenteriae

Conventionally raised crossbred Hampshire pigs were vaccinated orally with attenuated Treponema hyodysenteriae in combination with an anthelmintic, levamisole or dichlorvos. Pigs in group I (n = 9) were treated with levamisole and vaccinated with attenuated T hyodysenteriae and those in group II (n = 9) were treated with levamisole and permitted to commingle (contact exposure) with group I. Pigs in group III (n = 9) were vaccinated in a similar manner and were treated with dichlorvos. Pigs in group IV (n = 9) were treated with dichlorvos and permitted to commingle with group III. Control pigs (group V; n = 9) were not given any anthelmintic, nor were they vaccinated; they were housed separately. During the 8-week interval between vaccination and challenge inoculation, 4 total days and 8 total days of diarrhea were observed in pigs in groups I and II, respectively. Likewise, 5 total days and 10 total days of diarrhea were seen in groups III and IV, respectively. In all groups, the pigs tended to shed the organism in their feces after they were vaccinated or challenge inoculated, as determined by a fluorescent antibody technique (FAT) and culture procedure (CP). Overall mean shedding patterns of 5.5% and 24.5% identified by CP and FAT, respectively, were seen in the 2 levamisole-treated groups (I and II). In contrast, mean shedding patterns of 4% and 18% of the isolation attempts were detected by CP and FAT, respectively, in the 2 dichlorvos-treated groups. Diarrhea and shedding of T hyodysenteriae in the controls (group V) did not occur.(ABSTRACT TRUNCATED AT 250 WORDS)