Heterogeneity of Staphylococcus pseudintermedius isolates from atopic and healthy dogs

Staphylococcus pseudintermedius is part of the normal canine flora but frequently causes pyoderma in canine atopic dermatitis (AD). This study aimed to determine whether particular S. pseudintermedius strains were associated with AD and/or pyoderma. Ninety‐six S. pseudintermedius isolates from the ear, nares, perineum and lesions of 21 atopic and 16 healthy dogs were lysed with proteinase K and digested with 40 U SmaI. Restriction products were separated using pulsed‐field gel electrophoresis (PFGE) with an Oxford S. aureus control and lambda‐ladder DNA concatomer markers. A dendrogram was constructed by the unweighted pair group method. All isolates showed a ≥56% similarity coefficient. Nine distinct PFGE clusters were identified, as follows: five from both atopic and healthy dogs; three from atopic dogs only; and one from healthy dogs only. Nine clusters were isolated from the nares, eight from the perineum, five from the ears and six from pyoderma lesions. There were no significant differences in the frequency of isolation from atopic or healthy skin, body sites or infected lesions for any of the clusters. Two of six healthy dogs and 18 of 20 atopic dogs with multiple isolates had closely related isolates (less than three band differences) at more than one sampling site. Isolates from pyoderma lesions were closely related to at least one mucosal isolate in 11 of 16 dogs. Staphylococcus pseudintermedius isolates appear to be heterogeneous, and colonization or infection of atopic skin was not associated with any particular strain or cluster of strains.     

Establishment of native <Emphasis Type="Italic">Eucalyptus pauciflora</Emphasis> and exotic <Emphasis Type="Italic">Eucalyptus nitens</Emphasis> on former grazing land

The success of tree plantings in agricultural landscapes within dry and variable rainfall regions is often below expectations. The aims of this study were to investigate techniques for establishing a local native species (Eucalyptus pauciflora) and an exotic species (E. nitens) in the Midlands of Tasmania, a tree-denuded region that has been grazed for over 100 years in Australia. We established trials at the Sorrel Springs and Woodland Park properties. We compared six establishment techniques: control; understorey nurse planting; spot burn; mulch; tree-guard and; self-watering tree-guard. Three months, 1 and 6 years after planting we measured tree height, health index and mortality. At Sorrell Springs tree-guarded treatments had greater height than other treatments at year one, but there were no effects of treatment at year six on height, health or mortality. At Woodland Park mulch, burn and tree-guarded treatments had greater height 1 and 6 years after planting. At year six tree-guarded treatments had lower mortality than other treatments. At Sorrell Springs height of E. nitens was greater than E. pauciflora at year one but not at year six. At Woodland Park height of E. nitens was generally greater at year six but having almost double the incidence of mortality relative to E. pauciflora. We conclude that expensive tree-guarding is only warranted where high browse pressure is expected and that on such sites species with tough leaves that are resistant to browsing should be considered. Further that in environments marginal for growing trees, best practice establishment technique is generally as effective as additional treatments of understorey nurse planting, spot burning or mulching.     

Protection against bovine tuberculosis induced by oral vaccination of cattle with Mycobacterium bovis BCG is not enhanced by co-administration of mycobacterial protein vaccines

Mycobacterium bovis bacille Calmette-Guérin (BCG) delivered to calves by the oral route in a formulated lipid matrix has been previously shown to induce protection against bovine tuberculosis. A study was conducted in cattle to determine if a combination of a low dose of oral BCG and a protein vaccine could induce protective immunity to tuberculosis while not sensitising animals to tuberculin. Groups of calves (10 per group) were vaccinated by administering 2 × 10(7)colony forming units (CFU) of BCG orally or a combination of 2 × 10(7)CFU oral BCG and a protein vaccine comprised of M. bovis culture filtrate proteins (CFP) formulated with the adjuvants Chitin and Gel 01 and delivered by the intranasal route, or CFP formulated with Emulsigen and the TLR2 agonist Pam(3)CSK(4) and administered by the subcutaneous (s.c.) route. Two further groups were vaccinated with the CFP/Chitin/Gel 01 or CFP/Emulsigen/Pam(3)CSK(4) vaccines alone. Positive control groups were given 10(8)CFU oral BCG or 10(6)CFU s.c. BCG while a negative control group was non-vaccinated. All animals were challenged with M. bovis 15 weeks after vaccination and euthanized and necropsied at 16 weeks following challenge. Groups of cattle vaccinated with s.c. BCG, 10(8)CFU or 2 × 10(7)CFU oral BCG showed significant reductions in seven, three and four pathological or microbiological disease parameters, respectively, compared to the results for the non-vaccinated group. There was no evidence of protection in calves vaccinated with the combination of oral BCG and CFP/Emulsigen/Pam(3)CSK(4) or oral BCG and CFP/Chitin/Gel 01 or vaccinated with the protein vaccines alone. Positive responses in the comparative cervical skin test at 12 weeks after vaccination were only observed in animals vaccinated with s.c. BCG, 10(8)CFU oral BCG or a combination of 2 × 10(7)CFU oral BCG and CFP/Chitin/Gel 01. In conclusion, co-administration of a protein vaccine, administered by either systemic or mucosal routes with oral BCG did not enhance the protection conferred by administration of oral BCG alone.     

<Emphasis Type="Italic">Streptococcus uberis</Emphasis>-specific T cells are present in mammary gland secretions of cows and can be activated to kill <Emphasis Type="Italic">S. uberis</Emphasis>

The presence, phenotype and function of Streptococcus uberis-specific T cells in the mammary gland secretion (MGS) and blood of cows exposed to S. uberis were assessed. MGS T cells in the udder were purified and incubated with autologous blood monocytes as antigen-presenting cells (APC). Most cows, irrespective of prior S. uberis infection status and lactation status, were shown to have S. uberis-specific T cells both in MGS and in the blood. When cells from a subgroup of cows were studied, it was found that the S. uberis-specific T cells produced high levels of interferon-gamma (IFN-γ), but low levels of interleukin-10 (IL-10). A high percentage of responding T cells were of the CD8 ⁺ memory (CD45RO) subset. T cells from the MGS specific for S. uberis were propagated from animals during the drying off period and expanded in vitro using interleukin-2 (IL-2) and S. uberis antigens. This led to the accumulation of T cells of the CD8 ⁺ subset bearing memory cell markers (CD45A ⁻ , CD45RO ⁺ ), which released high levels of IFN-γ. Four of the five T cell lines derived from the MGS of three animals had substantial direct killing activity towards S. uberis in vitro. It is concluded that there is an emergence of S. uberis-specific bactericidal T cells in the MGS of cows after infection or environmental exposure to S. uberis. Vaccines aimed at activating and expanding this T cell population in the mammary glands of cattle may offer an avenue for the prevention of mastitis caused by S. uberis.     

Isolation and characterisation of equine influenza viruses (H3N8) from Europe and North America from 2008 to 2009

Like other influenza A viruses, equine influenza virus undergoes antigenic drift. It is therefore essential that surveillance is carried out to ensure that recommended strains for inclusion in vaccines are kept up to date. Here we report antigenic and genetic characterisation carried out on equine influenza virus strains isolated in North America and Europe over a 2-year period from 2008 to 2009. Nasopharyngeal swabs were taken from equines showing acute clinical signs and submitted to diagnostic laboratories for testing and virus isolation in eggs. The sequence of the HA1 portion of the viral haemagglutinin was determined for each strain. Where possible, sequence was determined directly from swab material as well as from virus isolated in eggs. In Europe, 20 viruses were isolated from 15 sporadic outbreaks and 5 viruses were isolated from North America. All of the European and North American viruses were characterised as members of the Florida sublineage, with similarity to A/eq/Lincolnshire/1/07 (clade 1) or A/eq/Richmond/1/07 (clade 2). Antigenic characterisation by haemagglutination inhibition assay indicated that the two clades could be readily distinguished and there were also at least seven amino acid differences between them. The selection of vaccine strains for 2010 by the expert surveillance panel have taken these differences into account and it is now recommended that representatives of both Florida clade 1 and clade 2 are included in vaccines.     

Influence of tillage and method of metam sodium application on distribution and survival ofVerticillium dahliae in the soil and the development of verticillium wilt of potato

The effect of plowing and deep-rip tillage, in combination with chemigation or shank injection of metam sodium, onVerticillium dahliae populations and disease development was assessed in two fields with differing soil types and potato rotations. Soil samples were collected on a geo-referenced basis at depths of 0 to 10 cm and 10 to 20 cm before tillage, after tillage, after chemical application, and before planting and assayed for the presence of the pathogen. Propagules ofV. dahliae were detected at 140 of 141 sites sampled prior to tillage. Most (74.4% in heavy, sandy loam; 63.1% in light, loamy sand) were concentrated in upper 10 cm of the soil profile. Plowing redistributed inoculum vertically while deep-rip tillage did not. In the non-chemical treated areas of both fields, theVerticillium population reached a maximum between 25 July and 8 August before declining to near pre-tillage levels. Overall, the population generally was lower in the field with heavy soil, higher organic matter content, and a 3-year crop rotation. Metam sodium appeared to be most effective when shank injected, as the levels of inoculum in both fields declined by 60% to 80% following this application method. Chemigation was ineffective in the lighter soil, but the inoculum density in the deep-rip tillage area of the field with the heavier soil declined by nearly 20% in the upper and 60% in the lower strata following this treatment. The number ofV. dahliae propagules at the 10- to 20-cm depth in the plowed area of the same field was reduced by 25% following chemigation, but remained unchanged in the upper strata. Wilt was reduced in both fields by as much as 50% with shank injection of metam sodium with concomitant increases in total yield, marketable yield and gross income. Increases in total yield were significant (P<0.05) for the main effect of chemical, in the plowed area of the field with the heavier soil type following shank injection. These data suggest that growers might benefit from altering their tillage and chemical application practices as part of an integrated approach to managing Verticillium wilt.     

The effect of ethylene and abscisic acid on symptom expression of bacterial ring rot in eggplant and potato

Studies were performed to determine the effect that plant hormones involved with plant senescence (i.e., ethylene and abscisic acid) and photoperiod have on disease development and symptom expression of bacterial ring rot (BRR) caused byCorynebacterium sepedonicum (Spieck & Kotth.) Skapt & Burkh. Potato plants were grown either under a long (14 hr.) or short (10 hr.) photoperiod, while eggplants were grown only under a short (10 hr.) photoperiod. Disease severity ratings of BRR were found to be significantly higher (P=0.05) on potato plants grown under a short photoperiod compared to a long photoperiod. Plant heights of BRR infected plants were found to be significantly greater under the long photoperiod. Endogenous levels of ethylene were found to be significantly (P=0.05) greater in inoculated potato plants grown under a long photoperiod than inoculated plants grown under a short photoperiod. Results suggest that the plant hormones ethylene and abscisic acid do not significantly affect the disease development and symptom expression of BRR.     

A rapid onset, post-capture muscle necrosis in the Norway lobster, Nephrops norvegicus (L.) from the West coast of Scotland

A post-capture, abdominal muscle necrosis of rapid onset has been identified in Norway lobsters, Nephrops norvegicus (L.), captured off the West coast of Scotland. Economic losses, as a result of the mortality of these animals in transport, were encountered by Scottish wholesalers during the summer and autumn of 1999. Affected animals show a characteristic whitening of individual muscle fibres and fibre bundles of the abdomen within hours of capture, with a progression towards complete opacity of the abdominal musculature within a number of days. The pathology causes a loss of the normal function of the abdomen; thus, preventing the normal 'tail flip' swimming. Electron microscopy failed to reveal any obvious causative agent but showed that affected tissue displayed a progressive disruption of sarcomere organization, loss of Z-line material, condensation of myofibrils and infiltration of necrotic regions by granulocytes. SDS–PAGE of affected muscle tissue showed that there was a great reduction of most of the major contractile proteins. The condition most closely resembles idiopathic or spontaneous muscle necrosis, a pathology previously reported from both wild and cultured crustaceans. Damage to the integument in conjunction with exposure to various stressors during and immediately after capture is the most likely cause of the pathology. The rapid onset of the pathology has implications for the post-capture handling procedure for N. norvegicus and their subsequent vivier transport to market. It may also be partially responsible for the high mortality rate of undersized N. norvegicus returned to the sea after capture and aerial emersion.