Adherence of Actinobacillus pleuropneumoniae serotype 1 to swine buccal epithelial cells involves fibronectin

The swine pathogen Actinobacillus pleuropneumoniae serotype 1 was investigated for its ability to adhere to swine, rat, and human buccal epithelial cells (BEC). The highest number of bacteria adhered was to swine BEC. This binding ability was affected by heating, extreme pH, treatment with sodium dodecyl sulfate, ethylenediamine tetraacetate, or periodate, and proteolysis, suggesting that cell-surface glycoproteins participate in adherence and that adherence is based mostly on ionic interactions. Mannose and swine fibronectin may play a direct role in this interaction. Convalescent-phase serum from naturally infected pigs inhibited the adhesion. There was a correlation between bacterial pathogenicity as well as host specificity and the capacity for adherence to swine BEC. Adhesion to swine BEC provides a convenient method to study in vitro the adherence of A. pleuropneumoniae and other pathogens of the pig respiratory tract.     

Estimating nocturnal ecosystem respiration from the vertical turbulent flux and change in storage of CO2

Micrometeorological measurements of nighttime ecosystem respiration can be systematically biased when stable atmospheric conditions lead to drainage flows associated with decoupling of air flow above and within plant canopies. The associated horizontal and vertical advective fluxes cannot be measured using instrumentation on the single towers typically used at micrometeorological sites. A common approach to minimize bias is to use a threshold in friction velocity, u_*, to exclude periods when advection is assumed to be important, but this is problematic in situations when in-canopy flows are decoupled from the flow above. Using data from 25 flux stations in a wide variety of forest ecosystems globally, we examine the generality of a novel approach to estimating nocturnal respiration developed by van Gorsel et al. (van Gorsel, E., Leuning, R., Cleugh, H.A., Keith, H., Suni, T., 2007. Nocturnal carbon efflux: reconciliation of eddy covariance and chamber measurements using an alternative to the u_*-threshold filtering technique. Tellus 59B, 397–403, Tellus, 59B, 307-403). The approach is based on the assumption that advection is small relative to the vertical turbulent flux (F_C) and change in storage (F_S) of CO₂ in the few hours after sundown. The sum of F_C and F_S reach a maximum during this period which is used to derive a temperature response function for ecosystem respiration. Measured hourly soil temperatures are then used with this function to estimate respiration R_Rmax. The new approach yielded excellent agreement with (1) independent measurements using respiration chambers, (2) with estimates using ecosystem light-response curves of F_c + F_s extrapolated to zero light, R_LRC, and (3) with a detailed process-based forest ecosystem model, R_cast. At most sites respiration rates estimated using the u_*-filter, R_ust, were smaller than R_Rmax and R_LRC. Agreement of our approach with independent measurements indicates that R_Rmax provides an excellent estimate of nighttime ecosystem respiration.     

Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences

Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions.     

Inoculation of ectomycorrhizal fungi contributes to the survival of tree seedlings in a copper mine tailing

To advance our understanding of the effects of inoculation with ectomycorrhizal fungi (EMF) on seedling colonization in mine wastelands, we conducted a field experiment in a copper tailing. Six-month-old seedlings of Japanese red pine (Pinus densiflora) and oak (Quercus variabilis) separately inoculated with three EMF species (Pisolithus sp., Cenococcum geophilum, Laccaria laccata) were transplanted to the copper tailing. The survival rates of tree seedlings were monitored monthly, and growth (biomass and height), contents of nutrients and heavy metals (K, P, Ca, Mg, Cu, Zn), and mycorrhizal infection rates of seedlings were determined 6 months after planting. Oak seedlings exhibited higher survival rates than pine seedlings after 6 months of growth on the tailing. EMF inoculations of pine seedlings significantly enhanced their survival, growth, and nutrient uptake. In contrast, EMF inoculations of oak seedlings improved growth only in terms of biomass. Additionally, EMF inoculation caused pine seedlings to accumulate more Cu and Zn in roots compared to non-inoculated seedlings, whereas inoculation inhibited the accumulation of heavy metals in shoots. However, similar results were not observed in oak seedlings. Observations of roots indicated that the rates of mycorrhizal infection of both tree species had dramatically declined at harvest time. In conclusion, ectomycorrhizal symbioses can improve the survival and performance of pine seedlings in mine tailings. The present study provided direct evidence of the importance of EMF inoculation of seedlings to the reforestation of mine wastelands.     

The vitamin A requirement and the vitamin A status of growing cattle. 1. Studies of calves

Five experiments with 18 to 36 male calves each of the black and white dairy cattle breed (age: 14-21 days, initial live weight: approximately 45 kg per animal) were carried out in order to investigate the influence of various vitamin A supply (0-80,000 IU per 100 kg LW and day) on dry matter intake and weight gain as well as the vitamin A status of liver and blood plasma over 84 days. The calves consumed a diet free of carotene and vitamin A consisting of milk replacer, concentrate and chopped wheat straw. The calves were fed in three experiments for a longer time in order to observe the further vitamin A depletion. Nine animals consumed an unsupplemented ration, nine other one got 10,000 IU vitamin A per 100 kg LW and day. Biopsies of liver and plasma samples were taken from 4 animals per group every four weeks. The various vitamin A supplementation did not significantly influence the dry matter intake (Mean: 1.67; 1.48 to 1.80 kg DM per animal and day) and the weight gain of calves (Mean: 702, 599 to 770 g per animal and day). First vitamin A deficiency symptoms (reduced feed intake, decreased weight gain, diarrhoea etc.) were observed in animals of unsupplemented group after 100 days of experiments. After 84 days the vitamin A concentration of liver of animals of unsupplemented groups decreased to 1.3-32.2% compared with the begin of experiments (60.6-155.7 mumol/kg fresh matter). Up to 51% of initial concentration were found when 10,000 IU vitamin A per 100 kg LW and day were fed. About 25,000 IU vitamin A per 100 kg LW and day were required in order to keep the initial level of vitamin A concentration of liver. The plasma vitamin A concentration is unsuitable for estimation of vitamin A status of calves. The concentration of vitamin A of liver and plasma amounted to 114 mumol per kg and 0.25 mumol per litre at the begin of experiments. The vitamin A concentration of liver of unsupplemented group decreased to 20 mumol per kg, that of plasma increased to 0.28 mumol per 1 at the end. A strong vitamin A deficiency (liver concentration: less than 10 mumol/kg) may cause a decrease of vitamin A concentration of blood.     

Allometric model of the height–diameter curve for even-aged pure stands of Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>)

We derived an allometric model of the height–diameter curve for even-aged pure stands, which was a modification of the earlier model proposed by Inoue (2000a). An individual-dependent allometric equation was used as the height–diameter curve. Using the discriminant analysis method, all trees composed of a stand were stratified into upper and lower trees. It was assumed that both relationships between mean tree height H _m and upper tree height H _u and between mean DBH D _m and mean DBH of upper trees D _u could be described by the time-dependent allometric power equations. The height–diameter curve showed an average relationship between tree height and DBH of a given stand at a given time, and hence it could be assumed that the height–diameter curve contained two points (D _m , H _m ) and (D _u , H _u ). With these assumptions, we derived an allometirc model of height–diameter curve, which allowed the coefficients of the curve to be estimated from mean tree height and mean DBH. The proposed model was fitted to Japanese cedar (Cryptomeria japonica D. Don) data. The error ratio of the allometric model ranged from 2.254% to 13.412% (mean = 6.785%), which was significantly smaller than that of the earlier model. When the error of mean tree height was ±1.0 m or less, the effect of the error of mean tree height on the error ratio was comparatively small. This suggested that the error of ±1.0 m in mean tree height could be accepted in the estimation of height–diameter curve using the allometric model. These features enable us to combine the allometric model with Hirata’s vertical angle-count sampling or growth models. In conclusion, the allometric model would be one of the most practical and convenient approaches for estimating the height–diameter relationship of even-aged pure stands.     

The relative plasma availabilities of ivermectin in reindeer (Rangifer tarandus tarandus) following subcutaneous and two different oral formulation applications

Background

Overwintering (breeding) reindeer (Rangifer tarandus tarandus) are commonly treated with ivermectin against parasitic infestations once yearly in autumn-winter roundups. The only preparations registered to reindeer are those for subcutaneous injection. However, also oral extra-label ivermectin administration is used. Twenty-six, 8-month-old reindeer calves were randomly allocated into three groups. Group 1 (n = 9) received oral ivermectin mixture (Ivomec® vet mixt. 0.8 mg/ml, oral ovine liquid drench formulation), Group 2 (n = 9) oral ivermectin paste (Ivomec® vet 18.7 mg/g equine paste), and Group 3 (n = 8) subcutaneous injection of ivermectin (Ivomec® 10 mg/ml vet inj.), each group at a dose of 200 μg/kg body weight. Blood samples were collected at treatment and at days 1, 2, 3, 6, 9 and 16 post treatment. Plasma concentrations of ivermectin were determined by high-pressure liquid chromatography (HPLC) with fluorescence detection.

Results

The peak plasma concentration (C_max) was reached by 2 days after each treatment. The C_max and Area Under Curve (AUC) differed significantly between the groups: C_max was 30.2 ± 3.9, 14.9 ± 5.7 and 63.1 ± 13.1 ng/ml, and AUC_∞ was 2881 ± 462, 1299 ± 342 and 6718 ± 1620 ng*h/ml for groups 1, 2 and 3, respectively (mean ± standard deviation).

Conclusions

The differences in plasma concentrations of ivermectin are concomitant with earlier observed differences in antiparasitic efficacy, which discounts the use of the equine paste in reindeer in favour of the oral ovine liquid drench formulation, or preferably, the reindeer-registered subcutaneous injection formulation.