T lymphocyte responses during early enteric Mycobacterium avium subspecies paratuberculosis infection in cattle

Johne's disease (JD) is a costly intestinal disease of ruminants caused by Mycobacterium avium subspecies paratuberculosis (Map), which is transmitted to perinatal calves by the fecal-oral route. Disease control efforts focus on identification and culling of infected cattle from herds; therefore failure to identify animals early is a major obstacle to reducing transmission. Development of host immunity during early JD remain incompletely characterized so detecting subclinical JD using immunologic techniques is a substantial challenge in the field. Development of a test with high sensitivity and specificity is a major research goal with significant implications for the cattle industry. The objectives of this study were to compare early Map-specific T lymphocyte responses in naive, experimentally Map infected and Map vaccinated calves using a subcutaneous matrigel biopolymer-based assay. We examined the phenotype of recruited lymphocytes and local interferon gamma (IFNγ) production within subcutaneously placed matrigel containing Map antigen 30 days after experimentally induced intestinal Map infection or Map vaccination. We show that IFNγ-secreting CD4+ T cells are recruited to matrigel sites in vaccinated but not infected or naïve calves. γδ T cells recruited to matrigel sites of Map-infected calves were mostly WC1-, while γδ T cells recruited to matrigel sites of Map-vaccinated calves were predominantly WC1+. IFNγ at matrigel sites was a discriminating factor between infected calves, naïve calves and vaccinated calves. These data contribute to our understanding of early anti-Map immunity, and may be useful for detecting early intestinal Map infections in calves or for enhancing our ability to discriminate between Map-infected and Map-vaccinated calves.     

Growth, survival and fillet composition of paddlefish, Polyodon spathula (Walbaum) fed commercial trout or catfish feeds

Paddlefish are gaining increasing acceptance as an aquaculture species worldwide. Commercial trout feeds, containing high protein and lipid levels, are currently used in intensive culture; however, nutritional requirements of paddlefish are not currently known. A study was conducted examining the effects on growth, survival and fillet composition of juvenile paddlefish when fed commercial feeds differing in protein and lipid levels. Paddlefish larvae were first stocked in 14.0 m³ round tanks and fed trout starter feeds for 43 days until trained to accept a 1.6 mm pellet. Paddlefish juveniles of mean weight (±SE) 20±0.27 g were randomly stocked into six0.02 ha ponds at 12 500 ha^?1 and fed floating commercial trout or catfish (lower protein and lipid) feeds, twice daily (08:00 and 15:30 hours) for 92–97 days. At harvest, there were no significant differences in final weight, percent survival, specific growth rate , relative growth and feed conversion ratio between treatments, which averaged 223.6 g, 96.2%, 2.5% day^?1, 10.2 and 1.98 respectively. Surface feeding activity index was significantly higher in ponds supplied with catfish feed than in ponds supplied with trout feeds. Relative pellet buoyancy was not a factor in feeding activity. Fulton's condition factor averaged0.238, was not significantly different, and was similar to a reported value for extensively cultured paddlefish (zooplanktivore). There was no significant difference in liver somatic index between treatments, which averaged 1.91%. Percent protein and moisture of fillets averaged 14.9% and 80.9%, respectively, and were not significantly different between treatments. However, lipid content of fillets was significantly higher in paddlefish fed the trout feed (4.45%), compared with paddlefish fed the catfish feed (2.42%). Fillet lipid content for both treatments was higher than reported values for extensively cultured paddlefish. Percent abdominal fat was significantly higher (0.82%) in paddlefish fed the trout feed compared with paddlefish fed the catfish feed (0.52%). Results from this study indicate that paddlefish can be fed a commercial catfish feed labeled to contain 32% protein and 4.5% lipid without adverse effects on growth, survival and fillet composition, lowering production costs.     

Assessing common bottlenose dolphin (Tursiops truncatus) population structure in Mississippi Sound and coastal waters of the north central Gulf of Mexico

1. Population structure of highly mobile marine organisms can be complex and difficult to study, but it is important to understand how populations partition themselves within their environment for accurate assessment of both natural and anthropogenic impacts and successful management. The 2010 Deepwater Horizon oil spill negatively impacted common bottlenose dolphins (Tursiops truncatus) within Mississippi Sound and the surrounding north central Gulf of Mexico (GOMx); however, little was known about their underlying population structure in these waters. Thus, it was unclear how many demographically independent populations were affected by the spill.
2. Common bottlenose dolphin samples were collected throughout inshore waters of Mississippi Sound and coastal waters of the north-central GOMx. Mitochondrial DNA control region sequence data and 19 nuclear microsatellite loci were analysed to determine how many populations are present and characterize their range throughout these waters.
3. Bayesian clustering and migration analyses identified two genetically distinct and demographically independent populations: one predominantly inhabiting Mississippi Sound and adjacent coastal waters, and a second population extending generally from offshore of Mobile Bay, Alabama, east along the Florida Panhandle. Neither of these populations align with the currently delineated management stocks previously used to estimate impacts from the oil spill on common bottlenose dolphins in this portion of the GOMx.
4. These results suggest that revisions may be necessary so that management stocks accurately represent the demographically independent populations present in these waters. Furthermore, better comprehension of underlying population structure will enhance impact assessments on common bottlenose dolphins and provide more appropriate baseline data to support future restoration and conservation objectives.

     

Technical note: isolation and characterization of sheep ruminal epithelial cells

A system for the isolation and characterization of sheep ruminal epithelial cells has been developed. Ruminal papillae from the ventral cranial sac of 12- to 24-wk-old Dorset ram lambs were subjected to serial tryptic digestions. Initial digestions (two 15-min periods) in the series were used to remove keratinized cells of the stratum corneum, which were able to produce only small amounts of beta-hydroxybutyrate from butyrate (5.37 +/- .72 nmol/120 min/per milligram of dry cell weight). Later digest fractions, containing primarily cells from the stratum basale, exhibited high viabilities (75 to 95%) and proved capable of converting butyrate to beta-hydroxybutyrate at relatively high rates (33.6 +/- 6.7 nmol/120 min per milligram of dry cell weight). Neither acetate nor propionate underwent significant conversion to beta-hydroxybutyrate. However, addition of acetate inhibited (77.4% of control) and addition of propionate stimulated (200% of control) beta-hydroxybutyrate production. Acetate addition reduced the propionate-induced stimulation of beta-hydroxybutyrate production from butyrate (136% of control). These results are similar to those obtained from in vitro incubations of ruminal papillae and suggest that an isolated cell system may prove useful in the further study of ruminal epithelial metabolism.     

Locating and protecting critical reserve sites to minimize expected and worst-case losses

There has been much recent interest in the development of systematic reserve selection methods that are capable of incorporating uncertainty associated with site destruction. This paper makes a contribution to this line of research by presenting two different optimization models for minimizing species losses within a planning region. Given limited acquisition budgets, the first minimizes expected species losses over all possible site loss patterns outside the reserve network while the second minimizes maximum species losses following the worst-case loss of a restricted subset of nonreserve sites. By incorporating the uncertainty of site destruction directly into the decision planning process, these models allow a conservation planner to take a less defensive and more strategic view of reserve selection that seeks to minimize species losses through the targeted acquisition of high-value/high-risk sites. We compare both of these methods to a more standard approach, which simply maximizes within reserve representation without regard for the varied level of threat faced by different sites and species. Results on a realistic dataset show that significant reductions in species losses can be achieved using either of these more intelligent modeling frameworks.     

Sequence variants in SLITRK1 are associated with Tourette's syndrome

Tourette's syndrome (TS) is a genetically influenced developmental neuropsychiatric disorder characterized by chronic vocal and motor tics. We studied Slit and Trk-like 1 (SLITRK1) as a candidate gene on chromosome 13q31.1 because of its proximity to a de novo chromosomal inversion in a child with TS. Among 174 unrelated probands, we identified a frameshift mutation and two independent occurrences of the identical variant in the binding site for microRNA hsa-miR-189. These variants were absent from 3600 control chromosomes. SLITRK1 mRNA and hsa-miR-189 showed an overlapping expression pattern in brain regions previously implicated in TS. Wild-type SLITRK1, but not the frameshift mutant, enhanced dendritic growth in primary neuronal cultures. Collectively, these findings support the association of rare SLITRK1 sequence variants with TS.     

Peptide binding and release by proteins implicated as catalysts of protein assembly

Two members of the hsp70 family, termed hsc70 and BiP, have been implicated in promoting protein folding and assembly processes in the cytoplasm and the lumen of the endoplasmic reticulum, respectively. Short hydrophilic (8 to 25 residues) synthetic peptides have now been tested as possible mimics of polypeptide chain substrates to help define an enzymatic basis for these activities. Both BiP and hsc70 have specific peptide binding sites. Peptide binding elicits hydrolysis of adenosine triphosphate, with the subsequent release of bound peptide.     

Diversity considerations in HIV-1 vaccine selection

Globally, human immunodeficiency virus-type 1 (HIV-1) is extraordinarily variable, and this diversity poses a major obstacle to AIDS vaccine development. Currently, candidate vaccines are derived from isolates, with the hope that they will be sufficiently cross-reactive to protect against circulating viruses. This may be overly optimistic, however, given that HIV-1 envelope proteins can differ in more than 30% of their amino acids. To contend with the diversity, country-specific vaccines are being considered, but evolutionary relationships may be more useful than regional considerations. Consensus or ancestor sequences could be used in vaccine design to minimize the genetic differences between vaccine strains and contemporary isolates, effectively reducing the extent of diversity by half.