Use of a polymerase chain reaction to subtype Mycobacterium avium subspecies paratuberculosis, an increasingly important pathogen from farmed deer in New Zealand

AIMS: To review the number of microbiologically-confirmed cases of Johne's disease in farmed deer since 2000, and determine the prevalence of the bovine and ovine subtypes of Mycobacterium avium subsp paratuberculosis (M. paratuberculosis), using a highly specific polymerase chain reaction (PCR) test on samples from infected herds. METHODS: The number of cases of M. paratuberculosis in farmed deer identified by culture or IS900 PCR was documented. A highly specific PCR test was applied to subtype M. paratuberculosis from BACTEC 12B cultures selected on the basis of one culture per deer herd, to give a wide coverage of herds in New Zealand. RESULTS: From January 2001 to October 2005, M. paratuberculosis was isolated from 1,141 farmed deer, and has now been identified by microbiological testing in over 600 deer herds in New Zealand. The bovine subtype of M. paratuberculosis was shown by a highly specific PCR test to be present in 91/95 herds examined; the ovine subtype was found in the remaining four herds. CONCLUSIONS: Since 2000, there has been a substantial increase in both the number of microbiologically-confirmed cases of Johne's disease in farmed deer and the number of infected herds. Johne's disease is now widespread and common in deer herds throughout New Zealand. Whilst the bovine subtype of M. paratuberculosis predominates in deer herds in New Zealand in which Johne's disease has been confirmed, the occasional finding of the ovine subtype highlights the need to consider both sheep and cattle as potential sources of infection for farmed deer.     

Effects of sodium hyaluronate and methylprednisolone acetate on proteoglycan metabolism in equine articular chondrocytes treated with interleukin-1

OBJECTIVE: To determine the effects of sodium hyaluronate (HA) in combination with methylprednisolone acetate (MPA) on interleukin-1 (IL-1)-induced inflammation in equine articular cartilage pellets. Sample POPULATION: Chondrocytes collected from 7 horses euthanatized for problems unrelated to the musculoskeletal system. PROCEDURES: Chondrocyte pellets were treated with medium (negative control); medium containing IL-1 (positive control); or medium containing IL-1 with MPA only (0.05 or 0.5 mg/mL), HA only (0.2 or 2 mg/mL), or MPA (0.05 or 0.5 mg/mL) and HA (0.2 or 2 mg/mL) in combination. Proteoglycan (PG) synthesis was determined by incorporation of sulfur 35-labeled sodium sulfate into PGs. Glycosaminoglycan (GAG) content of the media and the pellets and total pellet DNA content were determined. RESULTS: Methylprednisolone acetate at 0.5 mg/mL caused an increase in PG synthesis, whereas HA had no effect alone. The combination of MPA, both 0.05 mg/mL and 0.5 mg/mL, with HA at 2 mg/mL increased PG synthesis, compared with IL-1-treated control. All treatment groups containing the high concentration of MPA (0.5 mg/mL) and the high concentration of HA (2.0 mg/mL) had pellets with increased GAG content. The addition of HA caused an increase in total GAG content in the media, regardless of MPA treatment. Cyclooxygenase-2 mRNA and aggrecan mRNA expression was significantly reduced with MPA treatment. Total pellet DNA content was unchanged by any treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Our results indicate that MPA in combination with HA has beneficial effects on PG metabolism of IL-1-treated equine chondrocytes.     

Sorption and catalytic hydrolysis of diethatyl-ethyl on homoionic clays

Sorption and catalytic hydrolysis of the herbicide diethatyl-ethyl [N-chloroacetyl-N-(2,6-diethylphenyl)glycine ethyl ester] on homoionic Na(+)-, K(+)-, Ca(2+)-, and Mg(2+)-montmorillonite clays were studied in aqueous media. The Freundlich sorption coefficient, K(f), measured from isotherms on clay followed the order of Na(+) approximately K(+) > Mg(2+) approximately Ca (2+). Analysis of FT-IR spectra of diethatyl-ethyl sorbed on clay suggests probable bonding at the carboxyl and amide carbonyl groups of the herbicide. The rate of herbicide hydrolysis in homoionic clay suspensions followed the same order as that for sorption, indicating that sorption may have preceded and thus caused hydrolysis. Preliminary product identification showed that hydrolysis occurred via nucleophilic substitution at the carboxyl carbon, causing cleavage of the ester bond and formation of diethatyl and its dechlorinated derivative, and at the amide carbon, yielding an ethyl ester derivative and its acid. These pathways also suggest that hydrolysis of diethatyl-ethyl was catalyzed by sorption on the clay surface.     

Liquid chromatographic-fluorometric determination of cinnamyl alcohol in perfumes, colognes, and toilet waters

A liquid chromatographic (LC) method is described for the determination of cinnamyl alcohol (3-phenyl-2-propen-1-ol) in fragrance compositions. The fragrance product is partially cleaned up by diluting the fragrance with a 95% ethanol-water mixture and passing it through a short column containing RP-8 packing. An aliquot of the effluent is then analyzed by LC using an RP-18 column interfaced to a spectrophotofluorometer equipped with double monochromators. The fluorescence emission intensity of the eluted cinnamyl alcohol is measured and compared with that of a standard to calculate the amount of cinnamyl alcohol present. Recoveries from fragrance products fortified with cinnamyl alcohol at levels ranging from 0.0020 to 0.060 mg/mL ranged from 85 to 105% with a mean of 94%. The lowest level of determination was 0.0005 mg/mL.     

Organic matter effects on phase partition of 1,3-dichloropropene in soil

The fumigant 1,3-dichloropropene (1,3-D), in combination with chloropicrin, is considered a major replacement to methyl bromide (MeBr). This study was conducted to better understand phase partitioning of 1,3-D and the role of organic matter in its adsorption to soil. Partition of 1,3-D between air and water (K(H)), and between soil and water (K(f)), was determined by quantifying the concentration in both phases upon equilibrium. At 20 degrees C, the K(H) values of (Z)- and (E)-1,3-D were 0.052 and 0.033, respectively. In four California and Florida soils, the K(f) values of 1,3-D isomers ranged from 0.39 to 8.55, and the K(oc) values ranged from 18 to 60. The relatively high K(H) and low K(f) imply that 1,3-D is highly mobile in most soils after subsurface application. Adsorption of 1,3-D in native soils and soils amended with manure compost increased with increasing soil organic matter content. This suggests that organic wastes may be applied to soil to increase 1,3-D adsorption, thus reducing its potential for offsite movement.     

The United Kingdom national materials exposure programme

In 1986, the National Materials Exposure Programme was set up within the United Kingdom to investigate the effects of acid deposition on buildings and building materials. Thirty sites were chosen, which represented a range of geographical and pollution climates. Each site met a minimum meteorological and pollution monitoring regime (including SO₂, NO₂). After four years, other sites were included (with less frequent data collection) and some sites removed. At each site, samples of 3 types of stone, mild steel, painted steel, Cu, Al and galvanised steel were exposed, with some of the stone sheltered from direct precipitation. Samples were removed periodically for analysis and dose-response relations derived for different materials. The empirical relationships derived are in the form of: decay rate=a [SO₊] + b [H₂] + c [rainfall] + d These dose response relations have been used to develop critical load maps for materials for the United Kingdom. Eight years of data have been collected, some for the UNECE task force programme. Laboratory tests using an Atmospheric Flow Chamber were also undertaken. Since the beginning of the programme addition materials have been exposed on some sites including mortars. A further set of eight sites has been used to assess the effects of ozone on a range of organic materials (for example polyvinyl chloride, polycarbonate, sealants). The paper presents up-to-date findings for the programme and confirms the dominance of dry deposition of sulphur dioxide as the main decay process for sensitive materials in areas of significant pollution.     

Comparative population structure and reproductive biology of the critically endangered shrub Grevillea althoferorum and two closely related more common congeners

Grevillea althoferorum is a critically endangered, sprouting shrub known from two disjunct populations within the South-West Botanical Province of Western Australia. This study compares the conservation biology of G. althoferorum and two closely related but more common congeners, the non-sprouter G. rudis and the sprouter G. synapheae subsp. pachyphylla in order to determine whether there are differences in reproductive and ecological attributes that might explain why G. althoferorum is rare. In contrast to the more common species, neither population of G. althoferorum exhibited evidence of seedling recruitment. However, the northern population was confirmed to be clonal and was actively recruiting from root suckers. Both populations of G. althoferorum were found to have reduced amounts of viable pollen on stigmas in comparison with the other species. The fruit set at the southern population of G. althoferorum was considerably lower than that found for the common species, with only 0.15% of flowers setting fruit and no fruit was produced in the northern population. In addition no evidence of a soil seed bank was found for either population of G. althoferorum, but G. rudis and G. synapheae subsp. pachyphylla both had soil stored seed which germinated following treatment with aqueous smoke solution. Sexual recruitment at both populations of G. althoferorum was absent, and reproduction appears to be predominantly clonal in the northern population. Management strategies for G. althoferorum should therefore focus on the protection of adult plants from accidental destruction.     

Characterization by restriction endonuclease analysis and seroagglutination of strains of Mycobacterium avium and Mycobacterium intracellulare obtained from farmed deer.

Restriction endonuclease analysis and seroagglutination were used to characterize strains of Mycobacterium avium and Mycobacterium intracellulare (collectively, MAI) recovered from 1 local herd and 2 imported shipments of red deer (Cervus elaphus) that developed sensitization to bovine tuberculin during skin testing. A total of 31 MAI strains were isolated from lymph node pools (head, thorax, abdomen, and peripheral regions) of 21 of 29 local deer. Similarly, 15 MAI strains were isolated from the lymph node pools of 12 deer from the 2 imported shipments. Mycobacterial strains were isolated from more than 1 of the lymph node pools of 9 local and 2 imported deer. Most of the strains (59% from local deer, 46% from imported deer) were recovered from the lymph nodes of the head region. After restriction endonuclease analysis of these isolates using the enzymes Bcl I, BstEII, and Pvu II, 26 of the strains from the local herd were separated into 3 groups, each consisting of strains with indistinguishable or closely related patterns. Seroagglutination results indicated that the first of these groups contained strains belonging to serotype 1, the second group contained strains belonging to serotype 8; and the third group of strains belonged to serotype 3 and 9. The 5 remaining strains from the local herd had unrelated restriction patterns. One of these belonged to serotype 3, whereas the remaining 4 could not be serotyped. Restriction analysis of the 15 strains from the imported deer identified 2 groups. Seroagglutination results indicated that 1 group contained strains belonging to serotype 2 and the other group contained strains belonging to serotype 8.(ABSTRACT TRUNCATED AT 250 WORDS)     

Glial membranes at the node of Ranvier prevent neurite outgrowth

Nodes of Ranvier are regularly placed, nonmyelinated axon segments along myelinated nerves. Here we show that nodal membranes isolated from the central nervous system (CNS) of mammals restricted neurite outgrowth of cultured neurons. Proteomic analysis of these membranes revealed several inhibitors of neurite outgrowth, including the oligodendrocyte myelin glycoprotein (OMgp). In rat spinal cord, OMgp was not localized to compact myelin, as previously thought, but to oligodendroglia-like cells, whose processes converge to form a ring that completely encircles the nodes. In OMgp-null mice, CNS nodes were abnormally wide and collateral sprouting was observed. Nodal ensheathment in the CNS may stabilize the node and prevent axonal sprouting.     

Zilpaterol hydrochloride supplementation has no effect on the shelf life of ground beef

The objective of this study was to determine the effect of feeding zilpaterol hydrochloride (ZH) on the shelf life and stability of ground beef. Beef knuckles and plates were obtained from USDA Select beef heifer carcasses from control (CON) animals or those supplemented with ZH (8.33 mg/kg of dietary DM basis) for the last 20 d of the finishing period. Subprimals were coarsely ground and blended to produce an 80% lean product. The mixture was vacuum-stuffed into chubs and placed in dark storage at 2 to 4°C for 7, 14, or 21 d before fine grinding. Each week, the finely ground samples were packaged on expanded polystyrene trays overwrapped with polyvinyl chloride film and placed in refrigerated retail cases (0 to 2°C) under continuous fluorescent lighting to simulate retail display. Samples were subjected to a variety of analyses at different time intervals (h) during simulated display, including composition analysis, thiobarbituric acid-reacting substance analysis (TBA), sensory color, instrumental color, and aerobic plate count. Data analysis revealed trained sensory color and discoloration scores were similar between CON and ZH-treated samples. Instrumental L* and b* values for CON and ZH-treated samples did not differ (P = 0.13 and 0.19, respectively). Instrumental a* values declined (P < 0.05) over the display period for CON and ZH ground beef. However, a* values for ZH ground beef stored for 7 d were greater (P < 0.05) than CON values at 18 through 72 h of display. There was a treatment × storage day interaction (P < 0.001) for TBA values with ZH having smaller TBA values than CON after 7 d of dark storage. There was no difference (P = 0.21) in aerobic plate count between ZH and CON ground beef samples. Overall, ground beef from cattle supplemented with ZH was equal to or better than CON for sensory color and discoloration, instrumental color, and stability variables, including TBA reactive substances and aerobic plate counts.