Tumor Necrosis Factor-α, Interleukins-12(p40), 6, and 10 levels in cerebrospinal fluid and outcome prediction in Ossimi sheep with encephalitic listeriosis

Encephalitic listeriosis in sheep is a life-threatening disease. However, little is known about the cytokine response and their predictive value in this disease. The aim of present study was to assess the prognostic significance of Tumor Necrosis Factor-α (TNF-α), Interleukin-12(p40) (IL-12 p40), Interleukin-6 (IL-6), and Interleukin 10 (IL-10) levels in cerebrospinal fluid (CSF) in sheep with encephalitic listeriosis. Fifty-nine ewes in 14 flocks were diagnosed clinically as having listeriosis. CSF was collected and subjected to bacteriological examination and estimation of selected cytokines. Twenty-eight ewes were confirmed to be infected with Listeria monocytogenes. Based on antimicrobial sensitivity test, sheep were treated and the outcome was recorded as survivors (n = 10) and non-survivors (n = 18). Cutoff points for CSF cytokines were determined by Receiver operating characteristic analysis (ROC). Association between levels of CSF cytokines and outcome of listeriosis was assessed by logistic regression. TNF-α, IL-6 and IL-12(p40) levels as well as TNF-α/IL-10 ratio were significantly higher in non-survivors than survivors (p = 0.002, 0.0021, 0.0033, and 0.001, respectively). However, IL-10 level was significantly lower in non-survivors than survivors (p = 0.0058). ROC analysis revealed that IL-6 and TNF-α/IL-10 ratio had the highest AUC values (0.98, 0.984, respectively). Final multivariate logistic regression model showed that TNF-α/IL-10 ratio was the only variable that has predictive value for mortality in diseased sheep (p: 0.001; OR: 7.2; 95% CI: 5.7–9.8). TNF-α showed a positive correlation with IL-12β (r = 0.917) and IL-6 (r = 0.965). IL-12 (p40) showed also a positive correlation with IL-6 (r = 0.906). However, IL-10 showed a negative correlation with TNF-α (r = −0.915), IL-12(p40) (r = −0.790), and IL-6 (r = −0.902). In conclusion, TNF-α/IL-10 ratio may provide predictive information about outcome of encephalitic listeriosis in sheep.     

Evidence for the expression of two distinct MHC class II DRβ like molecules in the sheep

This study used monoclonal antibodies to sheep MHC class II molecules as well as an L cell transfectant (T8.1) which expresses DRA and DRB genes to show that two distinct DRβ chains are expressed in the sheep. Two anti-β chain specific monoclonal antibodies VPM37 and VPM43 react with DR antigen but not DQ antigen by ELISA. These two antibodies do not react with the DRβ chain expressed in the T8.1 cell line. Two-dimensional immunoblotting shows that these antibodies recognize a subgroup of the spots recognized by the DR-specific monoclonal antibody VPM57 which does react with the T8.1 β chain. Amino-terminal sequence analysis of the α chain associated with VPM37β chain shows that this α chain is homologous to the human DRα chain strongly indicating that the β chain is DR-like. VPM37 and VPM43 are shown to be directed against different epitopes on sheep MHC class II molecules so it is highly unlikely that the data can be explained by the presence of posttranslational modifications or the existence of a very common allele. These data provide clear evidence for the expression of two distinct DRP chains in the sheep.     

Multivariate immune defences and fitness in the wild: complex but ecologically important associations among plasma antibodies,health and survival

Despite our rapidly advancing mechanistic understanding of vertebrate immunity under controlled laboratory conditions, the links between immunity, infection and fitness under natural conditions remain poorly understood. Antibodies are central to acquired immune responses, and antibody levels circulating in vivo reflect a composite of constitutive and induced functional variants of diverse specificities (e.g. binding antigens from prevalent parasites, self tissues or novel non-self sources). Here, we measured plasma concentrations of 11 different antibody types in adult females from an unmanaged population of Soay sheep on St Kilda. Correlations among antibody measures were generally positive but weak, and eight of the measures independently predicted body mass, strongyle parasite egg count or survival over the subsequent winter. These independent and, in some cases, antagonistic relationships point to important multivariate immunological heterogeneities affecting organismal health and fitness in natural systems. Notably, we identified a strong positive association between anti-nematode immunoglobulin (Ig) G antibodies in summer and subsequent over-winter survival, providing rare evidence for a fitness benefit of helminth-specific immunity under natural conditions. Our results highlight both the evolutionary and ecological importance and the complex nature of the immune phenotype in the wild.     

Stock grazing of semi-natural habitats on National Trust land

Stock grazing is essential in many habitats owned by the National Trust if their nature conservation interests are to be maintained or enhanced. In this review, the benefits of grazing cattle, sheep and other livestock in different habitat types are described, together with constraints imposed by tenancy agreements and difficulties in obtaining the most appropriate animal. The majority of upland heaths, calcareous grasslands and lowland/coastal heaths on Trust land are being grazed satisfactorily. However, the nature conservation interest of parklands is currently being eroded in part because of inappropriate grazing regimes. It is argued that the best use of stock in the future is in extensive grazing regimes.     

Pronounced differences in the frequency of TaqI βA-inhibin alleles between sheep breeds with different reproductive performance

A total of 419 individuals of four breeds with differing fecundity (Rhoenschaf, Merinolandschaf, East Friesian Milksheep, Romanov) and several wild sheep of the Ovis musimon, O. orientalis, O. vignei and O. ammon groups were screened for genetic variation at the βA-inhibin (INHBA) locus with up to 11 enzymes. The four breeds differed significantly (P< 0.001) in TaqI allele frequencies. The frequency of the TaqI A allele coincided with the average litter size in each breed.     

An improved method for typing of plasma α1-protease inhibitor (PI1) variants in sheep

Summary
Sheep plasma α₁-protease inhibitor (PI1) variants were typed by analysing neuraminidase treated samples by isoelectric focusing in ultra-thin polyacrylamide gels followed by staining for trypsin inhibition. The 10 different PI1 pheno-types observed were shown to be controlled by four codominant alleles. The probability of sire exclusion provided by PI1 polymorphism was about 0.45 in both of the studied milk sheep breeds (Latxa and Karrantzar) of the Basque Country.     

Characterization and linkage mapping often sheep microsatellite markers derived from a sheep x hamster cell hybrid

A cosmid library has been constructed from a sheep x hamster cell hybrid containing sheep chromosome t₁, rob (6;24). Clones containing sheep DNA were identified by hybridizing to a total sheep genomic DNA probe. Small fragments (<500 bp) containing (AC)_n microsatellites were subcloned and sequenced. Ten microsatellite markers were characterized and six were mapped back to chromosomes 6 and 24. The remaining microsatellites mapped to chromosome 26, which was shown to be present in a small proportion of cells of the cell line.     

Biodistribution and Clearance of Intra-articular Liposomes in a Large Animal Model Using a Radiographic Marker

The intra-articular (IA) route of administration in treating arthritis has potential for targeting drug delivery to affected tissues, thereby minimising the attendant side-effects of systemically administered drugs. The ultra-structure of the synovium however facilitates rapid drug efflux from the joint; effectively the IA route is equivalent to other non-IV parenteral routes with regards absorption and redistribution into the systemic circulation. The aim of this study was to extend the drug residence time within the knee joint by using a liposome formulation. DPPC-based liposomes were prepared with the radio contrast agent iohexol as a drug marker. 8 sheep had their right knees injected IA with iohexol liposomes and the contralateral joints with either free iohexol or empty liposomes. Joints were radiographed at multiple time points up to 16 days post-injection. Iohexol-mediated radiopacity was quantified by densitometer. Sheep were sacrificed at the end of the study for microscopy of synovial tissues.

Good visualization of iohexol-mediated radiopacity with fine anatomical definition was possible throughout the experiment. Also evident on the films was extra-articular radiopacity with liposomes tracking along muscle facial planes. Cellular and tissue localization with light microscopy was possible through use of frozen sections and because of the large liposome size.

Residence of encapsulated iohexol within the knee joint was greatly prolonged.

Liposomal iohexol declined bi-exponentially with a terminal elimination half-life of 134 hours. In contrast, free iohexol was undetectable @ 3 hours post-injection.     

Non-genetic sources of variation of milk production and reproduction and interactions between both classes of traits in Sicilo-Sarde dairy sheep

This work aimed to study the sources of variation in productive and reproductive traits of the dairy Sicilo-Sarde ewes and to further investigate the interaction between both classes of traits. After edits, a database containing 5935 lactation records collected during 6 successive years in eight dairy flocks in the North of Tunisia was used. Total milked milk (TMM) in the milking-only period was retained as productive trait. The interval from the start of the mating period to the subsequent lambing (IML) and the lambing status (LS) were designed as reproductive traits. Sicilo-Sarde ewes had an average TMM of 60.93 l (±44.12) during 132.8 days (±46.6) after a suckling period of 100.4 days (±24.9). Average IML was 165.7 days. In a first step, the major factors influencing milk production and reproductive traits were determined. The significant sources of variation identified for TMM were: flock, month of lambing, year of lambing, parity, suckling length, litter size and milking-only length. Flock×month of the start of the mating period, parity, year of mating and litter size were identified as significant factors of variation for IML, while flock×month of the start of the mating period, parity and year of mating were identified as significant sources of variation for LS. In a second step, variance components were estimated using a three traits threshold mixed model, which combined LS as categorical trait and TMM and IML as continuous traits. Repeatability estimates were 0.21 (±0.03) for TMM, 0.09 (±0.02) for IML, and 0.10 (±0.05) for LS. Moreover, TMM and IML were found to be favorably associated for the flock× year of lambing effect (−0.45±0.18) but unfavorably associated for the animal effect (0.20±0.09).