Mimotope of Leptospira from phage-displayed random peptide library is reactive with both monoclonal antibodies and patients' sera

The study aim was to use random heptapeptide library displayed by bacteriophage T7 for identifying mimotopes from 15 monoclonal antibodies (MAbs) specific to Leptospira spp., and from four leptospirosis patient sera, respectively. The bound phages, selected from fourth round of bio-panning with each antibody, were cloned by plaque isolation and the binding specificity of individual clones were confirmed by enzyme-linked immunosorbent assay, before being further amplified and checked for phage peptide sequence using PCR and DNA sequencing. All together 150 phages were selected, mimotope from 86 phages (56.6%) were found to match with protein sequences of Leptospira from GenBank database. The predominant mimotopes were mimotope with sequence LTPCD that found in 27.3%, followed by TPCSK (16%), KSKKSS (4%), KTKRXAS (4%), SSKSYR (3.3%), DPNXNSF (3.3%), KSGRC (2.6%), TLINIF (2%), TPCI (2%), 1.33% each with mimotopes PKKS, PCNTKXTA, and CTKKK, and one phage each (0.66%) with mimotopes PTFGS, TNSKRK, SKSSRC, RSKRIR, VTNNTP, and CSNXSKR. Interestingly, mimotopes LTPCD, TPCSK, and TPCI were found to react with both MAb and patient's sera. The matched proteins from GenBank namely, leptospiral putative outer membrane protein (matched with mimotope PTFGS), thermolysin precursor protein (matched with mimotope TPCIXXGSAS), and hypothetical protein LIC12228 (matched with mimotope CSNXSKR), were found to locate at outer membrane of Leptospira. These phage mimotopes and matched proteins may have potential for further use as diagnostic reagent and immunogen against leptospirosis in the future. The results demonstrate that phage display technique has potential for rapidly identifying phage mimotopes that interact with leptospiral MAbs and patient's sera.     

Regionalisation of the muscle fascicle architecture in the equine longissimus dorsi muscle

REASON FOR PERFORMING STUDY: The longissimus dorsi is the largest muscle in the equine back and plays an important role in locomotor ability and performance in the horse. In vivo studies suggest that the mechanical function varies between different muscle segments, in part determined by anatomy. It is possible therefore that variations in function reflect variations in the anatomy of the longissimus dorsi along its length. OBJECTIVES: To identify if there are regional variations in muscle architecture of the longissimus dorsi. METHODS: Computed tomography scans were obtained from 8 cadaver backs to identify the cross-sectional area and volume of the muscle. The 3D orientations of the muscle fascicles were quantified in situ by their direction cosines using a magnetic digitisation system. RESULTS: Mean +/- s.e. volume of the muscle was 2.34 +/- 0.18 l for each side and mean length of the vertebral column from T4 to L6 dorsal spinous processes was 56.57 +/- 2.0 cm. There were significant differences in the cross-sectional area and the moment arm lengths between muscle segments (P<0.05). In the lumbar region, the mediolateral moment arms were 2-3 times greater than the dorsoventral moment arms. There were significant variations in the direction cosines (n = 7129) between muscle segments and between different regions within each segment (P<0.05). POTENTIAL RELEVANCE: These observations indicate that the muscle fascicle architecture predisposes the longissimus dorsi to different functions both along its length and between different regions within each segment. Detailed 3D anatomical measures of the structure with in vivo measures of function (back motion and muscle activity) will be investigated in further studies.     

Seasonal influences on quantitative changes in sweat-associated anatomy in native and thoroughbred horses

Stereological techniques were used to assess seasonal influences on morphometric characteristics of hair follicles, sweat and sebaceous glands in abattoir pelts of ponies (PN), thoroughbred (TB) and non-thoroughbred (NTB) horses. Volume density of sweat glands increased significantly from winter (0.061) to summer (0.098) in TB, and showed no change in NTB and a positive tendency in PN. There might be a body surface area : volume effect for sweat gland parameters as PN had smaller values than either TB or NTB, probably attributable to control of heat loss in winter. In summer, the skin remained thick and the volume density of sebaceous glands was increased in NTB, in contrast to TB where both were decreased. It is possible that in summer, sebum has a particular importance in NTB to enhance wicking of sweat through the pelt. TB showed significantly higher volume measurements of sebaceous glands than NTB and PN for winter: sebum has probably a special importance for water-proofing in TB in winter. PN showed no significant seasonal changes in sebaceous glands, but had a thinner summer skin. Winter values for hair follicle volume density between equine groups were similar (TB, NTB 0.066; PN 0.059), as was skin thickness (1.14-1.19 mm). The volume density lowered significantly in summer in TB and NTB. The volume of hair follicles under a unit area of skin surface decreased significantly in TB and nonsignificantly in NTB and PN. The seasonal adaptations of the skin shown here were most pronounced in TB and differed between breeds.     

Population genetic structure and genetic diversity of <Emphasis Type="Italic">Jatropha curcas</Emphasis> germplasm as investigated by 5′-anchored simple sequence repeat primers

Jatropha (Jatropha curcas L.) is widely distributed in the tropics with a promise for use as an oil crop for biodiesel. Understanding population genetic structure and diversity of J. curcas is important for germplasm collection and planning for breeding programs. Twelve 5′-anchored simple sequence repeat primers were selected and used for evaluation of population genetic structure and genetic diversity of J. curcas accessions collected from different regions within Thailand and introduced from other countries. A moderate level of genetic variation was found in all accessions studied. Cluster analysis did not reflect geographical distance in general as revealed by the Mantel test. An analysis of molecular variance showed that the majority (83.3%) of genetic diversity was within populations, indicating a high level of overlap and little population subdivision with respect to geographic area. An absence of subpopulation structure among neighboring regions may be caused by exchanging germplasm with vegetative propagation. This study provides a basis for breeders on selection of parental material to maximize genetic polymorphism in J. curcas breeding programs.     

Yeast mixture of liquid beer and cassava pulp with rice straw for the growth of dairy heifers

This study was conducted to determine the effects of mixtures of liquid brewer’s yeast (LBY) and cassava pulp (CVP) with rice straw (RS) on feed intake, digestibility, rumen fermentation, and growth of dairy heifers. Sixteen Holstein crossbred heifers (13.8?±?1.6 months old, 210?±?23 kg body weight (BW)) were randomly allocated to four feeding treatments with four replications, which were 0:0:100 (RS), 0:70:30 (0%LBY), 20:50:30 (20%LBY), and 50:20:30 (50%LBY), respectively, for LBY/CVP/RS on a fresh matter basis. The heifers were offered conventional concentrate at 1.5% initial body weight daily and fed the treatment diets ad libitum. Average daily gain and feed intake were not significantly different among the treatments. The heifers fed 50%LBY had the highest crude protein (CP) intake and DM, OM, and CP digestibility (P?<?0.05). The ruminal pH did not differ significantly among treatments, while NH₃-N was the highest (P?<?0.05) in 50%LBY. Total volatile fatty acid (VFA) concentrations and the molar proportion of each VFA were not significantly different among the treatments. Blood urea nitrogen concentrations of 50%LBY were the highest among the treatments (P?<?0.05). The results indicated that 50%LBY improved CP digestibility.