Evaluation of dietary addition of 2 essential oils from Achillea moschata,or their components (bornyl acetate,camphor, and eucalyptol) on in vitro ruminal fermentation and microbial community composition

This study investigated the effects of 2 Achillea moschata essential oils extracted from plants collected in 2 different valleys of the Italian Alps and 3 pure compounds of oils — bornyl acetate (BOR), camphor (CAM), and eucalyptol (EUCA) — on in vitro ruminal fermentation and microbiota. An in vitro batch fermentation experiment (Exp. 1) tested the addition of all of the substances (2 essential oils and 3 compounds) in fermentation bottles (120 mL) at 48 h of incubation, whereas a subsequent in vitro continuous culture experiment (Exp. 2) evaluated the pure compounds added to the fermenters (2 L) for a longer incubation period (9 d). In both experiments, total mixed rations were incubated with the additives, and samples without additives were included as the control (CTR). Each treatment was tested in duplicate and was repeated in 3 and 2 fermentation runs in Exp. 1 and 2, respectively. Gas production (GP) in Exp. 1 was similar for all of the treatments, and short chain volatile fatty acid (SCFA) production was similar in both experiments except for a decrease of SCFA produced (P = 0.029) due to EUCA addition in Exp. 2. Compared to CTR, BOR and CAM reduced the valerate proportion (P = 0.04) in Exp. 1, and increased (P < 0.01) the acetate proportion in Exp. 2. All treatments increased (P < 0.01) total protozoa counts (+36.7% and +48.4% compared to CTR on average for Exp. 1 and 2, respectively). In Exp. 1, all of the treatments lowered the Bacteroidetes and Firmicutes and increased the Proteobacteria relative abundances (P < 0.05), whereas in Exp. 2, the EUCA addition increased (P = 0.012) the Ruminococcus. In Exp. 1, methane (CH₄) as a proportion of the GP was lowered (P = 0.004) by the addition of CAM and EUCA compared to CTR, whereas in Exp. 2, EUCA reduced the amount of stoichiometrically calculated CH₄ compared to CTR. Overall, essential oils extracted from A. moschata and the pure compounds did not depress in vitro rumen fermentation, except for EUCA in Exp. 2. In both experiments, an increase of the protozoal population occurred for all the additives.     

Effects of intertrochanteric varus osteotomy on Norberg angle and percent coverage of the femoral head in displastic dogs

This study was conducted to assess the effects of femoral varus osteotomy on joint congruency in dogs affected by early stage hip dysplasia. Preoperative planning to move the femoral head within the acetabulum was carried out. Varisation of the femoral inclination angle (fIA) was achieved by Intertrochanteric Osteotomy (ITO). Norberg angle (NA), percent coverage (PC) of the femoral head by the acetabulum and fIA was measured from preoperative, immediate postoperative and first and second recheck radiographs of seven dogs that underwent an ITO (joint n = 9). There was significant (p < 0.05) improvement of both NA and PC in all patients as indicated by a change in the mean ± standard deviation of 78.9° ± 7.5 and 36.9% ± 5.2 to 92.2° ± 6.7 and 50.6% ± 8.3, respectively. No significant difference (p < 0.05) was observed between the values of the planned femoral inclination angle (pfIA) of the femur and the effective femoral inclination angle (efIA) obtained after surgery (115.9° ± 2.5 and 111.3° ± 6.4, respectively). These findings could encourage the use of ITO in veterinary practice and indicate that intertrochanteric varus osteotomy should be re-considered for the treatment of early stage hip dysplasia in dogs with radiological signs of joint incongruency.     

Ultrastructure of immature and mature human oocytes after cryotop vitrification

In vitro maturation of vitrified immature germinal vesicle (GV) oocytes is a promising fertility preservation option. We analyzed the ultrastructure of human GV oocytes after Cryotop vitrification (GVv) and compared it with fresh GV (GVc), fresh mature metaphase II (MIIc) and Cryotop-vitrified mature (MIIv) oocytes. By phase contrast microscopy and light microscopy, the oolemmal and cytoplasmic organization of fresh and vitrified oocytes did not show significant changes. GVv oocytes showed significant ultrastructural alterations of the microvilli in 40% of the samples; small vacuoles and occasional large/isolated vacuoles were abnormally present in the ooplasm periphery of 50% of samples. The ultrastructure of nuclei and mitochondria-vesicle (MV) complexes, as well as the distribution and characteristics of cortical granules (CGs), were comparable with those of GVc oocytes. MIIv oocytes showed an abnormal ultrastructure of microvilli in 30% of the samples and isolated large vacuoles in 70% of the samples. MV complexes were normal, but mitochondria-smooth endoplasmic reticulum aggregates appeared to be of reduced size. CGs were normally located under the oolemma but presented abnormalities in distribution and matrix electron density. In conclusion, Cryotop vitrification preserved main oocyte characteristics in the GV and MII stages, even if peculiar ultrastructural alterations appeared in both stages. This study also showed that the GV stage appears more suitable for vitrification than the MII stage, as indicated by the good ultrastructural preservation of important structures that are present only in immature oocytes, like the nucleus and migrating CGs.     

Effects of a selection of additives on in vitro ruminal methanogenesis and in situ and in vivo NDF digestibility

The effects of 18 essential oils, yeast, Quebracho tannin and Quillaja saponin on ruminal methane and gas production (GP) were studied in vitro. A lactating cow diet was incubated with rumen inoculum. Doses of the additives (mg/L) were: essential oils = 500; tannin = 67; yeast = 8.35 and 16.7; and saponin = 300. Lemongrass, estragole, eugenol, geraniol, limonen, thyme oil and thymol produced less gas (overall mean 33.8 mL/200 mg dry matter (DM)) than control (43.6 mL/200 mg DM; P < 0.001). Methane produced (mL/200 mg DM) by guaiacol (10.7), lemongrass (9.6), limonene (11.4), thyme oil (10.9) and thymol (2.1) was lower than control (12.5) (P < 0.001). Methane percentage on total GP was lower (P < 0.001) for cinnamic alcohol (25.4), guaiacol (24.5), thymol (19.7) and vanillin (26.3) than control (28.8). In a second experiment, thymol, guaiacol and yeast were added to the diet of dry fistulated cows to determine in situ neutral detergent fiber digestibility (NDFD) of six forages and in vivo dietary NDFD. Thymol and yeast decreased in situ NDFD after 24 h (33.9% and 33.5% vs. 38.1%; P = 0.008). Thymol decreased in vivo total tract NDFD (40.8% vs. 51.4%; P = 0.02). Differences in GP and methane levels were registered within classes of additives. A careful selection of additives may allow for the manipulation of ruminal fermentation.     

Partially defatted black soldier fly larva meal inclusion in piglet diets: effects on the growth performance,nutrient digestibility,blood profile,gut morphology and histological features

Background: The aim of this trial was to investigate the effects of different inclusion levels of a partial y defatted black soldier fly(BSF, Hermetia il ucens L.) larva meal on the growth performance, nutrient digestibility, blood profile, gut morphology and histological features of piglets. A total of 48 newly weaned piglets were individual y weighed(initial body weight(IBW): 6.1 ± 0.16 kg) and randomly al ocated to 3 dietary treatments(4 boxes as replicates/treatment and 4 animals/box). BSF larva meal was included at increasing levels(0% [BSF0], 5% [BSF5] and 10% [BSF10]) in isonitrogenous and isoenergetic diets formulated for two feeding phases: I(from d 1 to d 23) and I(from d 24 to d 61). The weight gain(WG), average daily gain(ADG), average daily feed intake(ADFI) and feed conversion ratio(FCR) were calculated for each feeding phase and for the whole trial. The haematochemical parameters and nutrient digestibility of the piglets were also evaluated. A total of 3 piglets per box were slaughtered on d 61 and the slaughtered piglets were submitted to morphometric investigations and histopathological examinations.Results: No overal significant differences were observed for growth performance(P 0.05), except for the ADFI of phase I, which showed a linear response to increasing BSF meal levels(P 0.05, maximum for the BSF10 group). Dietary BSF meal inclusion did not significantly influence the blood profile, except as far as monocytes and neutrophils are concerned,and these showed a linear and quadratic response, respectively, to increasing BSF meal levels(P 0.05, maximum for the BSF10 and BSF5 groups, respectively). On the other hand, the nutrient digestibility, gut morphology and histological features were not affected by dietary BSF meal inclusion(P 0.05).Conclusions: The obtained results show that a partial y defatted BSF larva meal can be used as a feed ingredient in diets for weaned piglets without negatively affecting their growth performance, nutrient digestibility, blood profile, gut morphology or histological features.     

Untersuchungen zum gezielten Einsatz von Insektiziden gegen den Rapsglanzkäfer (<Emphasis Type="Italic">Meligethes aeneus</Emphasis>) in der landwirtschaftlichen Praxis des Landes Brandenburg im Zeitraum von 2006 bis 2014

Before 2006, agents of class II pyrethroids were used to control rape pests including the rape pollen beetle mainly. Thus, the selection of pyrethroids-resistant pollen beetle populations was encouraged. The aim of our study was to identify ways to combat these populations. Therefore, data of 465 real on-farm insecticide applications from nine years since 2006 were analyzed with regard to their effects on the pollen beetle and own experiments were carried out to test the efficacy of approved insecticides and control strategies. The analysis showed that the highest efficiencies were obtained with agents of the group of organophosphates and with the active substances indoxacarb and pymetrozine. Beyond that, the data examined revealed that the efficiency of thiacloprid (an insecticide of the class of neonicotinoids) has remained relatively constant throughout the observation period. In contrast, the increase in pyrethroid-resistant pollen beetle populations reflects the further decrease in efficacy of class II pyrethroids in the period 2011–2014.In summary, it should be noted that currently available insecticides achieve a maximal abatement level of 80–85?%. Therefore, an economic damage cannot be prevented on rapeseed locations with a very strong pollen beetle infestation even if multiple use of insecticides happens.To prevent further escalation of the resistance problem, another aspect should be given greater attention in the future. 55?% of the insecticide applications in the practice dataset where carried out before the control threshold (BRW) for weakly developed oilseed rape crops was achieved; often in the stage of micro-bud (BBCH 51). In compliance with the currently suggested higher control threshold, only 15?% of the insecticide applications would have been necessary. Thus, a reduction of the number of insecticide applications could be achieved by considering the higher thresholds. This would significantly reduce the selection pressure. Therefore, plant protection advice should mediate between exploiting the control thresholds and using the most appropriate insecticide.     

Investigation of Genomic Variability of Xanthomonas Arboricola Pv. juglandis by AFLP Analysis

Xanthomonas arboricola pv. juglandis is the causal agent of walnut blight, one of the most important and widespread diseases of Persian (English) walnut (Juglans regiaL.), causing severe damage to leaves, twigs and nuts. To investigate the genomic variability of X. arboricolapv. juglandis, 66 isolates obtained from different countries (England, France, Italy, The Netherlands, Romania, Spain, USA, and New Zealand) were analysed using the Amplified Fragment Length Polymorphism (AFLP) technique. EcoRI and MseI were used as restriction endonucleases. Primers with a core sequence including endonuclease recognition sites and a 3^prime-terminal cytosine selective base for MseI primer, or no selective base for EcoRI primer, were used. Data were analysed by means of a multiple correspondence analysis. A total of 76 amplified polymorphic DNA fragments were used to compute relationships among isolates. The AFLP profiles of X. arboricola pv. juglandis isolates appeared to be reliably distinguishable from X. arboricola pv. pruni and X. arboricola pv. corylina, and from other Xanthomonas species, i.e. X. campestris pv. campestris, X. fragariae, X. hortorum, X. axonopodis pv. vesicatoria. Though this pathogen is associated with one single host genus, a high level of genomic diversity was found. This diversity might be partly explained by the geographic origin. Nevertheless, isolates with different patterns were collected within one country, and similar molecular patterns were found in isolates collected at different sites. However, genetic diversity might be influenced by exchanging vegetative material from different countries. Mixing of X. arboricola pv. juglandis isolates might have partly concealed the influence of the geographic location from which the bacteria were isolated.     

Activity of adjuvants in ‘aliette’ against Botryotinia fuckeliana (botrytis cinered) and their cross-resistance with benzimidazole fungicides

‘Aliette’, a formulation of the fungicide fosetyl-aluminium, is moderately active in vitro against the ascomycete Botryotinia fuckeliana (Botrytis cinerea). In contrast, fosetyl-aluminium and related chemicals, such as phosphonic acid and sodium phosphonate, have low efficacy, although it is enhanced by media with low phosphate content. This indicates that the effectiveness of ‘Aliette’ against B. fuckeliana is not due to fosetyl-aluminium but to adjuvants included in the formulation. The genetic basis of differences in sensitivity to ‘Aliette’ was investigated in field and laboratory isolates of B. fuckeliana. Genetic analysis showed that reduced sensitivity to ‘Aliette’ always co-segregated with resistance to benzimidazole fungicides in meiotic progeny of sexual crosses. This indicates that differences in sensitivity to ‘Aliette’ in B. fuckeliana are due to the major gene Mbcl of resistance to benzimidazole fungicides and active adjuvant(s) present in ‘Aliette’ are linked to benzimidazoles by cross-resistance.     

Characterization and genetic analysis of field isolates ofBotryotinia fuckeliana (Botrytis cinerea) resistant to dichlofluanid

Field isolates ofBotryotinia fuckeliana were collected from naturally infected plants. Their responses to the multisite fungicide dichlofluanid in mycelium growth test fell into three phenotypic classes, characterized by the following EC₅₀ (and MIC) values ing ml^–1: sensitivity, 1–3 (6–10); low resistance, 3–10 (> 100); high resistance, 10–30 (> 100). The corresponding values obtained for these classes in a spore germination test were respectively: 0.05 (0.2), 0.05–0.1 (0.5), 0.5–1 (0.9–1.5). Resistant isolates were crossed with two sensitive and two resistant strains of appropriate mating type to determine the genetic basis of resistance. Distribution of resistance phenotypes in ascospore progeny indicated that a gene, namedDic1, was probably responsible for the low or high resistance of 14 mutants selectively collected from experimental plots of greenhouse-grown gerbera sprayed several times with dichlofluanid or tolyfluanid. A second gene, namedDic2, was probably responsible for the low resistance displayed by two isolates (from grapevine and from carnation) maintained in the laboratory collection. As a result of the investigation, the use of dichlofluanid in integrated management programmes against grey mould is discussed.