The kinetics of cytokine production and CD25 expression by porcine lymphocyte subpopulations following exposure to classical swine fever virus (CSFV)

Surface expression of IL-2R-alpha (CD25) is widely used to identify activated lymphocyte populations, while interferon-gamma (IFN-gamma) levels have been shown to be a good indicator of cell-mediated immunity (CMI) in pigs. To investigate the relationship between these two parameters, we developed an intracellular cytokine-staining assay and studied the kinetics of cytokine (IFN-gamma and interleukin-10, IL-10) production relative to CD25 expression in porcine lymphocyte subpopulations, following immunization with a classical swine fever (CSF) vaccine. The number of activated memory T cells (CD4(+)CD8(+)CD25(+) cells) increased slightly in the peripheral blood mononuclear cell (PBMC) population soon after vaccination, then diminished within a few weeks. The number of activated cytotoxic T cells (CD4(-)CD8(+)CD25(+) cells) peaked approximately 2 weeks after the memory population. Although the number of IFN-gamma producing cells detected in this experiment was relatively low, the CD4(+)CD8(+) T cells were major IFN-gamma producers in the PBMCs throughout the experiment. In another experiment, CSF-vaccinated pigs were challenged with a virulent classical swine fever virus (CSFV), and the kinetics of CD25 expression and cytokine productions were monitored. Following exposure to the virus, the number of IFN-gamma producing cells in the PBMCs increased markedly in both the vaccinated and unvaccinated groups. The CD4(-)CD8(+) cells were major IFN-gamma producing cells in vaccinated pigs, while both CD4(+)CD8(+) and CD4(-)CD8(+) populations contributed to the IFN-gamma production in the control group. Interestingly, the enhanced IFN-gamma production was not associated with the upregulation of CD25 expression following the CSFV challenge. In addition, exposure to the virulent CSFV significantly increased interleukin-10 production by the CD4(-)CD8(+) populations in PBMCs of the unvaccinated pigs. Taken together, our results indicated that CD25 expression and IFN-gamma production were not tightly associated in porcine lymphocytes. In addition, the CD4(-)CD8(+) lymphocytes of the PBMCs played a major role in cytokine productions following the CSFV challenge.     

Production of eosinophil chemoattractant activity by ovine gastrointestinal nematodes

Eosinophilia is a well documented feature of helminth infections but the precise nature of the interaction between parasite and eosinophil remains an enigma. This paper describes experiments demonstrating that ruminant gastrointestinal trichostrongyles produce potent chemoattractant activity for ovine bone marrow-derived eosinophils in vitro. This activity was initially identified as a constituent of whole worm extracts of third and fourth larval (L3, L4), and adult stages of Teladorsagia circumcincta, and adult Haemonchus contortus. Similar activity was detected in excretory/secretory (E/S) material derived from live T. circumcincta L3. Subsequently, by adapting the assay technique to incorporate live worms directly into the system, it was shown that L3 of both T. circumcincta and H. contortus produced eosinophil chemoattractant activity. In contrast, neither whole worm extracts, or E/S preparations from mixed stages of the free-living nematode Caenorhabditis elegans contained eosinophil chemoattractant activity, and there was no evidence of chemoattractant production by live C. elegans. The results described are challenging to the traditional dogma that eosinophils are host-protective effector cells, and raise the intriguing possibility that ovine nematodes actively encourage recruitment of eosinophils. Local eosinophil-mediated mucosal damage, comparable to that seen in the asthmatic lung, may then provide a permissive local microenvironment for the parasite. Moreover, if they prove important for pathogenicity, nematode chemoattractants could offer future potential as novel therapeutic targets.     

Urinary tract infections-microbial virulence determinants and reactive oxygen species

The human urinary tract is able to combat with the microbial invasion under normal circumstances. To cause urinary tract infection the organism has to evade the host defense mechanisms by possessing distinct properties which contribute to the virulence of the organism hence called virulence determinants Ninety percent of uncomplicated urinary tract infections are caused by Escherichia coli, hence the knowledge of the virulence determinants of this organism can be extrapolated to other uropathogenic organism as well. Virulence determinants of uropathogenic E. coli include adhesins, siderophore production, polysaccharide coating, hemolysin production, outer membrane proteins etc. The intestinal E. coli, which are the reservoir of E. coli for causing UTI, lack these virulence determinants. On the other hand these virulence determinants enable the organism to colonize and invade the urinary tract. In addition these are important in acquiring the nutrients in other wise nutrient deficient environment. Further, they also help the organisms in triggering an inflammatory response and hence bringing about pathological changes which leads to symptomatic UTI. Severity of symptomatic infections and tissue damage during the infective process depends upon the magnitude of the inflammatory response triggered by the uropathogen which in turn is dependent upon the amount of extrcellular release of reactive oxygen species by the phagocytic cells; hence role of antioxidants as an adjunct to antibiotics in the treatment of infective process needs to be evaluated further.     

Oral malignant melanoma – the effect of coarse fractionation radiotherapy alone or with adjuvant carboplatin therapy

A retrospective study was undertaken of dogs presented to the Animal Health Trust for treatment of oral malignant melanoma, without radiographic evidence of pulmonary metastases. Group 1 (n = 13) received radiotherapy of the primary and any lymph node metastases (4 weekly fractions of 9 Gy); and group 2 (n = 15) were treated the same but additionally received between two and six doses carboplatin at 300 mg m^?2 every 3 weeks. Median survival times for the two groups were 307 and 286 days, respectively (P > 0.05). In addition, carboplatin therapy did not significantly reduce the proportion of dogs dying due to metastases (three from group 1 and four from group 2). We found no evidence of a beneficial effect of carboplatin therapy over radiotherapy alone.     

Growth performance, energy, and nitrogen balance of weanling pigs fed a cereal-based diet supplemented with Chinese rhubarb

Forty Swiss Large White piglets of 4 weeks of age were used to determine the effect of dietary Chinese rhubarb on growth performance, as well as on energy and nitrogen metabolism by means of indirect respiration calorimetry studies. A cereal-based diet, supplemented with four different dosages of Chinese rhubarb (0.0%, 0.25%, 0.5%, 1%), was offered ad libitum to the piglets. Gaseous exchange was measured, and urine, faeces, and blood samples were taken. The addition of 0.25% dietary rhubarb increased feed intake (+32%) and daily weight gain (+67%) compared to the diet containing the greatest amount of rhubarb (1%). The piglets fed diet supplemented with 0.25% rhubarb digested and metabolised more energy and nitrogen relative to the animals of treatment 1%. As a consequence, the same group (0.25%) converted more energy and nitrogen into body protein than piglets receiving diet supplemented with 1% rhubarb. However, compared to the control group, the addition of 0.25% rhubarb to the diet resulted in not significantly alter growth performance, energy, and nitrogen metabolism of the animals.Blood parameters were not affected by the rhubarb supplementation. The dose-responding effect of Chinese rhubarb was reflected by the dry matter content of faecal samples. According to the laxative effect of higher doses of rhubarb, the addition of 1% rhubarb to the diet reduced dry matter content of faeces. This effect was well visible in practice, although not relevant in statistical terms. The addition of 0.5% dietary rhubarb had no relevant effect on growth performance or on energy and nitrogen metabolism of treated animals.     

The effects of ractopamine hydrochloride on lean carcass yields and pork quality characteristics

One hundred eighty barrows were evaluated to determine the effects of ractopamine hydrochloride (RAC) on lean carcass yields and pork quality. The pens were blocked by weight (six pens per block) with starting block weights of 69.0, 70.7, 73.8, 76.6, 78.4, and 84.3 kg. Pens within a block were assigned randomly to one of three RAC treatments so each treatment in a block was replicated twice. Treatments (as-fed basis) included control diet, 10 ppm of RAC added (R10), and 20 ppm of RAC added (R20) and ranged from 25 to 41 d depending on block. Pigs were slaughtered by blocks when block average live weights were 109 kg. Gain and feed efficiency were improved (P < 0.05) with increasing dietary concentrations of RAC, but feed intake did not differ (P > 0.05). Dressing percentage was higher (P < 0.05) for RAC-treated pigs. Subjective color, firmness, marbling scores, and Minolta L* reflection of the LM were not different (P > 0.05) among treatments. Carcass weights were heavier (P < 0.05) for pigs treated with RAC compared with control pigs and were higher for R20 than for R10. The RAC-fed pigs had greater (P < 0.05) yields (actual and percentage of HCW) of the following Institutional Meat Purchase Specification (IMPS) cuts than control pigs: trimmed, boneless ham (IMPS-402C and IMPS-402G), loin (IMPS-414), sirloin, and Boston butt (IMPS-406A). Pigs treated with RAC had a greater (P < 0.05) percentage of fat-free lean trimmings (IMPS-418) than did control pigs. Pigs treated with the R20 concentration had increased (P < 0.05) water-holding capacity compared with control pigs. Purge loss decreased linearly (P < 0.05) with increasing RAC compared with control for 14-d aged, non-enhanced loins. Warner-Bratzler shear (WBS) force values measured for nonenhanced chops were greater for RAC-treated pigs than for control pigs with a low dose response (P = 0.001). Enhanced chop (salt and phosphate injection) WBS values did not differ (P > 0.05) among dietary treatments. Trained sensory evaluation panel results for tenderness decreased in a low-dose plateau response fashion for nonenhanced chops (P = 0.004). Tenderness of enhanced chops decreased linearly (P = 0.04) with increasing RAC concentrations. No differences (P > 0.05) were found in juiciness or flavor of enhanced or nonenhanced chops. Feeding RAC to late-finishing swine resulted in faster growing, more efficient animals with increased boneless subprimal yields, and it had little effect on pork juiciness and flavor.     

A new single nucleotide polymorphism in CAPN1 extends the current tenderness marker test to include cattle of Bos indicus, Bos taurus, and crossbred descent

The three objectives of this study were to 1) test for the existence of beef tenderness markers in the CAPN1 gene segregating in Brahman cattle; 2) test existing CAPN1 tenderness markers in indicus-influenced crossbred cattle; and 3) produce a revised marker system for use in cattle of all subspecies backgrounds. Previously, two SNP in the CAPN1 gene have been described that could be used to guide selection in Bos taurus cattle (designated Markers 316 and 530), but neither marker segregates at high frequency in Brahman cattle. In this study, we examined three additional SNP in CAPN1 to determine whether variation in this gene could be associated with tenderness in a large, multisire American Brahman population. One marker (termed 4751) was associated with shear force on postmortem d 7 (P < 0.01), 14 (P = 0.015), and 21 (P < 0.001) in this population, demonstrating that genetic variation important for tenderness segregates in Bos indicus cattle at or near CAPN1. Marker 4751 also was associated with shear force (P < 0.01) in the same large, multisire population of cattle of strictly Bos taurus descent that was used to develop the previously reported SNP (referred to as the Germplasm Evaluation [GPE] Cycle 7 population), indicating the possibility that one marker could have wide applicability in cattle of all subspecies backgrounds. To test this hypothesis, Marker 4751 was tested in a third large, multisire cattle population of crossbred subspecies descent (including sire breeds of Brangus, Beefmaster, Bonsmara, Romosinuano, Hereford, and Angus referred to as the GPE Cycle 8 population). The highly significant association of Marker 4751 with shear force in this population (P < 0.001) confirms the usefulness of Marker 4751 in cattle of all subspecies backgrounds, including Bos taurus, Bos indicus, and crossbred descent. This wide applicability adds substantial value over previously released Markers 316 and 530. However, Marker 316, which had previously been shown to be associated with tenderness in the GPE Cycle 7 population, also was highly associated with shear force in the GPE Cycle 8 animals (P < 0.001). Thus, Marker 316 may continue to be useful in a variety of populations with a high percentage of Bos taurus backgrounds. An optimal marker strategy for CAPN1 in many cases will be to use both Markers 316 and 4751.     

Carcass composition, bone mechanical properties, and meat quality traits in relation to growth rate in rabbits

The objective of this study was to characterize correlated responses in carcass composition, bone mechanical properties, and meat quality indicators to divergent selection for growth rate. Rabbits from low (LOW) or high (HIGH) lines divergently selected over five generations on 63-d BW and a control group (CONT) were used. Rabbits were slaughtered at an average BW of 2,306 +/- 65 g, corresponding to 63, 58, or 52 d of age in LOW (n = 41), CONT (n = 43), and HIGH (n = 44) groups, respectively. Postweaning ADG and G:F increased (P < 0.001) in the order LOW < CONT < HIGH groups. The lengths of the tibia and femur in rabbit legs decreased (P < 0.001) with increasing growth rate. Tibia and femur bone intrinsic stiffness, as assessed by a three-point flexure test, followed the order (P < 0.001) of HIGH < CONT < LOW groups. At the same BW, HIGH rabbits and CONT rabbits showed many similarities for dressing percent, carcass composition, and color and chemical composition of muscles or meat parts. In contrast, carcass yield and the relative proportion of hind part were 3% greater (P < 0.01) in LOW rabbits than in CONT and HIGH rabbits. The meat-to-bone ratio in the hind leg was 11% greater (P < 0.001) in the LOW group compared with the CONT and HIGH groups. Yellow color index and moisture content in LM, a fast-twitch glycolytic muscle, were lower (P < 0.05) in LOW rabbits than in HIGH rabbits, but ultimate pH, WHC, and cooking loss did not differ between the two growth-selected lines. Ultimate pH in semitendinosus, a mixed slow- and fast-twitch oxidoglycolytic muscle, was less (P < 0.001) in HIGH rabbits than in CONT and LOW rabbits. However, lactate dehydrogenase and isocitrate dehydrogenase activities in this muscle (n = 21 per group), assessed at the time of slaughter as markers of glycolytic and oxidative capacities, respectively, did not differ among groups. Growth rate did not modify mean cross-sectional area and type frequency of myofibers in semitendinosus muscle. Finally, Warner-Bratzler shear force and total energy needed to achieve rupture did not differ among groups in semitendinosus muscle (n = 21 per group), whereas they were the least (P < 0.001) in LM (n = 128) from HIGH rabbits. Results from this study indicate that improvement of growth rate by selection has little effect in rabbit carcass and meat quality traits.     

Lignin characteristics of bast fiber and core in kenaf, bark and wood of paper mulberry and mulberry

The structural features of bast fiber and core lignins in kenaf (Hibiscus cannabinus), bark and wood lignin of paper mulberry (Broussonetia papyrifera (L.) Vent × Broussonetia kazinoki Sieb.) and mulberry (Morus bombycis) were characterized by alkaline nitrobenzene oxidation, ozonation and methoxyl group determination. Bj?rkman lignins were isolated from bast fiber and core, and bark and wood fractions of the plant samples, and structural characteristics were investigated by ¹H NMR and ¹³C NMR spectroscopies. Kenaf bast fiber gave very high molar ratio of syringaldehyde to vanillin (S/V) of alkaline nitrobenzene oxidation products, while methoxyl content was about the same as that of the core fraction. Results of ¹H NMR and ¹³C NMR of Bj?rkman lignin suggested the presence of aliphatic fragments in lignins isolated from paper mulberry and mulberry bark, but not in kenaf bast fiber. The lower yield of alkaline nitrobenzene oxidation products from bast fiber and bark might be due to the higher content of condensed structure of lignin compared to core fraction. Total yield of erythronic (E) and threonic (T) acids of ozonation products and the molar ratio of erythronic acid to threonic acid (E/T) of the bast fibers and bark were lower than the corresponding core and wood fractions, suggesting that the contents of arylglycerol-β-aryl ether intermonomer linkages in the bast fiber and bark lignin were lower than those of the core and wood fractions. Methoxyl content of bark lignin was lower than the corresponding wood lignin. The methoxyl content of the extract-free kenaf bast fiber was similar to that of the core fraction, while the values of paper mulberry and mulberry bark were about one-half of the corresponding wood fractions, respectively. In bark lignins, the methoxyl contents of Klason lignin and Bj?rkman lignin from bark were lower than those of the extract-free barks. This result suggests that the purity of Klason lignin and Bj?rkman lignins of bark may be rather low.