Kinetics and intrapulmonary disposition of tilmicosin after single and repeated oral bolus administrations to rabbits

Tilmicosin (TIM, Pulmotil®) was administered to eight rabbits by oral gavage at a dose of 12.5 mg/kg body weight for 2, 5, and 7 days, and its plasma kinetics and intrapulmonary disposition were investigated. TIM concentrations in plasma samples collected after days 1 and 6 of treatment were measured by high-performance liquid chromatography with ultraviolet detection. The pharmacokinetic parameters, obtained by non-compartmental analysis of TIM plasma concentrations, did not show any significant variations between days 1 and 6. From the second day of treatment, TIM concentrations attained in lung tissue and pulmonary alveolar macrophages (PAM) exceeded those in plasma by 7- and 400-fold, respectively, and high levels were maintained in lung tissues during the entire treatment duration. After the first day of withdrawal, a fast decline in TIM levels in both plasma and lung tissue was observed, but in PAM, much higher concentrations were maintained after 3 days of TIM withdrawal.     

Weed control modifies Tuber melanosporum mycelial expansion in young oak plantations

Context

Black truffle (Tuber melanosporum) cultivation is a promising agro-forestry alternative for Mediterranean rural areas, but adequate weed control at seedling establishment still remains a challenge in black truffle plantations.

Aims

The aim of this study is to evaluate the effects of several weed control strategies on early development of Quercus ilex seedlings and the symbiotic T. melanosporum.

Materials and methods

In a young black truffle-inoculated holm oak plantation, we assessed for 3 years the effects of two types of mechanical weed control and five mulches in a young Q. ilex plantation inoculated with T. melanosporum. Herbaceous cover, seedling growth and abundance of T. melanosporum mycelium, based on PCR analysis of soil DNA extracts using T. melanosporum-specific primers, were estimated to determine the effectiveness of these treatments in controlling weeds and supporting the growth of both the host tree and the target fungus.

Results

The amount of T. melanosporum mycelium in the soil 30 cm around the seedlings was larger under double-layer white mulch than in the rest of treatments tested. Under the white colour mulches, which had the largest light reflection, we registered the cooler soil temperature, and the best weed control was observed on the single- and double-layer black truffles and double-layer white mulch.

Conclusion

The effects of double-layer white mulch on herbaceous cover, soil temperature, reflected light, and the expansion of T. melanosporum bring us closer to being able to substitute traditional tilling of truffle orchards for the less expensive mulching treatments.     

Main biotic drivers of tree growth in a developing Juniperus thurifera stand in central Spain

Over the last few decades, abandonment of traditional management practices in Spain has led to widespread stand densification and has favoured the expansion of some forest species that previously exhibited more restricted ranges. Spanish juniper (Juniperus thurifera L.) woodlands are experiencing this phenomenon due to agricultural land abandonment and a decrease in the livestock pressure. Yet the main drivers underlying stand structure and dynamics at this novel scenario are poorly understood. In this study, we investigate the main biotic drivers of tree growth in a high-density stand of the dioecious J. thurifera at an early developmental stage (mean tree age of 32, ca. 50 years after land abandonment). Tree growth was measured by coring 299 individuals of different reproductive classes (male, female and non-reproductive). Neighbourhood models were used to assess the relative importance of tree size and neighbourhood competition on tree growth of each reproductive class in the study plot. We found that tree size had the strongest effect on tree growth, whereas the effect of intraspecific competition was negligible. We observed differences in growth patterns among reproductive classes along trunk diameter sizes. Thus, at smaller sizes, the three reproductive classes presented identical patterns of growth. However, at bigger sizes, females were the fastest growing individuals, followed by males and non-reproductive individuals. Overall, our results suggest that in young J. thurifera monospecific forests, where self-thinning processes may have not undergone yet, tree size and the reproductive class could play a relatively more important role than competition as drivers of tree growth. These findings constitute new information which contributes to understanding growth dynamics at early developmental stages in this dioecious species. Furthermore, our results provide guidelines for silvicultural managing, suggesting that at these young juniper stands thinning would likely not translate into enhanced growth on remnant trees.     

Immunization with Eimeria ninakohlyakimovae-live attenuated oocysts protect goat kids from clinical coccidiosis

Caprine coccidiosis, affecting mainly young goat kids around the weaning period, is worldwide the most important disease in the goat industry. Control of caprine coccidiosis is increasingly hampered by resistances developed against coccidiostatic drugs leading to an enhanced need for anticoccidial vaccines. In the current study we conducted an oral immunization trial with live attenuated sporulated Eimeria ninakohlyakimovae oocysts. Sporulated E. ninakohlyakimovae oocysts were attenuated by X-irradiation technique. The experimental design included a total of 18 goat kids divided into the following groups: (i) animals immunized with attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-irradiated homologous oocysts (group 1); (ii) animals infected with non-attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-attenuated homologous oocysts (group 2); (iii) animals primary-infected with untreated E. ninakohlyakimovae oocysts at 8 weeks of age (control of the challenge infection, group 3); (iv) non-infected control animals (group 4). Goat kids immunized with live attenuated E. ninakohlyakimovae oocysts (group 1) excreted significantly less oocysts in the faeces (95.3% reduction) than kids infected with non-attenuated ones (group 2). Furthermore, immunization with live but attenuated oocysts resulted in ameliorated clinical coccidiosis compared to goat kids infected with untreated oocysts (group 2) and resulted in equally reduced signs of coccidiosis after challenge infection compared to acquired immunity driven by non-attenuated oocysts. Overall, the present study demonstrates for the first time that live attenuated E. ninakohlyakimovae oocysts orally administered showed almost no pathogenicity but enough immunogenicity in terms of immunoprotection. Importantly, vaccinated animals still shed low amounts of oocysts, guaranteeing environmental contamination and consecutive booster infections to sustain ongoing immunity.     

Electroencephalographic Responses to a Noxious Surgical Stimulus in Mules,Horses, and Ponies

The aim of this study was to investigate the electroencephalographic (EEG) response of equidae to a castration stimulus. Study 1 included 11 mules (2½–8 years; 230–315 kg) and 11 horses (1½–3½ years; 315–480 kg); study 2 included four ponies (15–17 months; 176–229 kg). They were castrated under halothane anesthesia after acepromazine premedication (IV [study 1] and intramuscular [study 2]) and thiopental anesthetic induction. Animals were castrated using a semiclosed technique (study 1) and a closed technique (study 2). Raw EEG data were analyzed and the EEG variables, median frequency (F50), total power (Ptot), and spectral edge frequency (F95), were derived using standard techniques at skin incision (skin) and emasculation (emasc) time points. Baseline values of F50, Ptot, and F95 for each animal were used to calculate percentage change from baseline at skin incision and emasculation. Differences were observed in Ptot and F50 data between hemispheres in horses but not mules (study 1) and in one pony (study 2). A response to castration (>10% change relative to baseline) was observed in eight horses (73% of animals) and four mules (36% of animals) for F50 and nine horses (82%) and four mules (36%) for Ptot. No changes in F95 data were observed in any animal in study 1. Responses to castration were observed in three ponies (75% of animals) for F50, one pony (25%) for F95, and all ponies for Ptot. Alteration of acepromazine administration and castration technique produced a protocol that identified changes in EEG frequency and power in response to castration.     

Kinetic study of the oxidation of quercetin by mushroom tyrosinase

The kinetic behavior of mushroom tyrosinase in the presence of the flavonol quercetin was studied. This flavonol was oxidized by mushroom tyrosinase and the reaction was followed by recording spectral changes over time. The spectra obtained during the reaction showed two isosbectic points, indicating a stable o-quinone. When quercetin was oxidized by tyrosinase in the presence of cysteine and 3-methyl-2-benzothiazolone hydrazone (Besthorn's hydrazone, MBTH) isosbestic points were also observed indicating a definite stoichiometry. From the data analysis of the initial rate in the presence of MBTH, the kinetic parameters: = (16.2 +/- 0.6) microM/min, = (0.12 +/- 0.01) mM, (/) = (V(max)/K(S)(')()) = (13.5 +/- 1.4) x 10(-)(2) min(-)(1), = (6.2 +/- 0.6) s(-)(1) were determined. We propose that quercetin acts simultaneously as a substrate and a rapid reversible inhibitor of mushroom tyrosinase, depending on how it binds to the copper atom of the enzyme active site. Thus, if the binding occurs through the hydroxylic groups at the C3' and C4' positions, quercetin acts as a substrate, while if it occurs through the hydroxylic group at the C3 position of the pyrone ring, quercetin acts as an inhibitor.     

Mycoflora of the 'green water' culture system of tiger shrimp Penaeus monodon Fabricius

This study was conducted to quantify and characterize the mycoflora associated with the ‘green water’ culture system of Penaeus monodon. Samples of water, tilapia gut and mucus, and shrimp hepatopancreas from three shrimp farms were collected during 15, 30, 45 and 60 days of culture (DOC). Results showed that high fungal loads were observed in tilapia gut (total: 117–1352 colony forming unit (CFU) 5 cm hind gut^?1; yeasts: 0–136 CFU 5 cm hind gut^?1) and mucus (total: 12–311 CFU (5 cm²)^?1; yeasts: 0–88 CFU (5 cm²)^?1), while minimal fungal populations were observed in water samples (total: 0–110CFU mL^?1; yeasts: 0–5 CFU ml^?1). Shrimp hepatopancreas harboured a very low number of filamentous fungi (0–27 CFU 0.1 g^?1) and yeasts (0–7CFU 0.1 g^?1) especially at 60 DOC. The filamentous fungal isolates were dominated by Penicillium and Aspergillus species, while the yeast populations were dominated by Rhodotorula and Saccharomyces species. The dominance of these fungi on tilapia mucus and gut and their presence in the rearing water might play an important role in the overall mechanisms involved in the control of luminous Vibrio in the ‘green water’ grow‐out culture of P. monodon.     

At which spatial scale are plant species composition and diversity affected in beech forests?

• Context  

Landscape structure is crucial for forest conservation in regions where the natural forest is fragmented. Practical conservation is currently shifting from local stands to a landscape perspective, although few studies have tested the relative effects of different spatial scales on plant species composition and diversity in forests.     

Potential synergistic effect of Melia azedarach fruit extract and Beauveria bassiana in the control of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in cattle infestations

The use of a concentrate emulsion of Melia azedarach green fruits and a suspension of the fungus Beauveria bassiana was evaluated in the control of Rhipicephalus microplus on artificially infested cattle. The evaluation was conducted following the protocol established by the Brazilian Agriculture Ministry. Five groups of 4 or 5 animals were allocated to one of the following treatments: emulsion concentrate of M. azedarach at 0.25% (T AZED 0.25%), emulsion concentrate of M. azedarach at 0.5% (T AZED 0.5%), B. bassiana at 2.4 × 10(8) conidia (T BASS), association of the concentrate of M. azedarach at 0.25% with B. bassiana at 2.4 × 10(8) conidia (T AZED 0.25%+BASS), and control (untreated). The association of the two compounds provided better results than any one isolated treatment, indicating compatibility or perhaps a synergy between M. azedarach and B. bassiana. This treatment resulted in fewer engorged females (129 ± 70) than in the control group (233 ± 82), showing high performance against all developmental stages of the tick. Results revealed an apparent synergistic effect of M. azedarach and B. bassiana in the control of R. microplus that should be further investigated.     

Bacterial community structure and detection of putative plant growth-promoting rhizobacteria associated with plants grown in Chilean agro-ecosystems and undisturbed ecosystems

Soil microorganisms with phytase- and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activities are widely studied as plant growth-promoting rhizobacteria (PGPR). Here, we explored the bacterial community structure and occurrence of putative PGPR in plants grown in agro-ecosystems and undisturbed ecosystems from northern, central, and southern Chile. Total rhizobacterial community structure was evaluated by denaturing gradient gel electrophoresis, and dominant bands present in diverse ecosystems were sequenced. Significant differences in total bacterial communities were shown with some bacterial orders (Enterobacteriales, Actinomycetales, and Rhizobiales) being highly similar to both ecosystems. Twenty-nine putative PGPR, showing phytate- and ACC-degrading activities and production of auxin, were selected from across the sites. Based on 16S rRNA gene sequencing, the putative PGPR were characterized as Enterobacteriales (Enterobacter, Serratia, Pantoea, Rahnella, Leclercia), Pseudomonas, and Bacillus, consistent with previously reported PGPR and endophytic bacteria. Beta-propeller phytase genes with similarity to Bacillus were also identified. PGPR from agro-ecosystems appeared to show higher auxin production compared to those from undisturbed ecosystems. This study demonstrates that putative PGPR are widely distributed across Chilean soils. Further understanding of their contribution to the growth and adaptation of plant hosts to local soil conditions may provide opportunity for development of new PGPR in Chilean agriculture.