Actinobacillus pleuropneumoniae is impaired by the garlic volatile allyl methyl sulfide (AMS) in vitro and in-feed garlic alleviates pleuropneumonia in a pig model

Decomposition products of ingested garlic are to a certain extent excreted via the lungs. If the supposed health-supporting capacities associated with garlic extend to these exhaled sulfurous compounds, they could have an effect on the course of pneumonia. In this study, the garlic-derived volatile allyl methyl sulfide (AMS) as a lead compound of volatile garlic metabolites was shown to exhibit an antibacterial effect against the pig pathogen Actinobacillus pleuropneumoniae serotype 9. AMS caused a delay in the appearance of the optical density-monitored growth of A. pleuropneumoniae in medium when compared to unaffected growth curves, yet without lowering the stationary phase yield at the concentration range tested. At 1.1mM, AMS impaired the in vitro growth rate of A. pleuropneumoniae serotype 9 by 8% compared to unimpeded growth. In an animal trial, a garlic-fed group of 15 pigs that received a diet with 5% garlic feed component and a control group of 15 pigs that received a diet without garlic were infected with A. pleuropneumoniae serotype 2 via an aerosol and subsequently followed for 4 days. At the day of the challenge, blood AMS in the garlic-fed group amounted to 0.32 ± 0.13 μM. A beneficial, alleviating effect of garlic on the course and severity of an A. pleuropneumoniae infection in pigs was indicated by the reduced occurrence of characteristic pleuropneumonia lesions (27% of the lungs affected in the garlic-fed group vs. 47% in the control group) and a near to significant (p=0.06) lower relative lung weight post mortem in the garlic-fed group.     

A two year BTV-8 vaccination follow up: molecular diagnostics and assessment of humoral and cellular immune reactions

The compulsory vaccination campaign against Bluetongue virus serotype eight (BTV-8) in Germany was exercised in the state of Bavaria using three commercial monovalent inactivated vaccines given provisional marketing authorisation for emergency use. In eleven Bavarian farms representing a cross sectional area of the state the immune reactions of sheep and cattle were followed over a two year period (2008-2009) using cELISA, a serum neutralisation test (SNT) and interferon gamma (IFN-γ) ELISPOT. For molecular diagnostics of BTV genome presence two recommended real time quantitative RT-PCR protocols were applied. The recommended vaccination scheme led to low or even undetectable antibody titers (ELISA) in serum samples of both cattle and sheep. A fourfold increase of the vaccine dose in cattle, however, induced higher ELISA titers and virus neutralising antibodies. Accordingly, repeated vaccination in sheep caused an increase in ELISA-antibody titers. BTV-8 neutralising antibodies occurred in most animals only after multiple vaccinations in the second year of the campaign. The secretion of interferon gamma (IFN-γ) in ELISPOT after in vitro re-stimulation of PBMC of BTV-8 vaccinated animals with BTV was evaluated in the field for the first time. Sera of BTV-8 infected or vaccinated animals neutralising BTV-8 could also neutralise an Italian BTV serotype 1 cell culture adapted strain and PBMC of such animals secreted IFN-γ when stimulated with BTV-1.     

Antioxidant and biocidal activities of Carum nigrum (seed) essential oil, oleoresin, and their selected components

In the present study, chemical constituents of the essential oil and oleoresin of the seed from Carum nigrum obtained by hydrodistillation and Soxhlet extraction using acetone, respectively, have been studied by GC and GC-MS techniques. The major component was dillapiole (29.9%) followed by germacrene B (21.4%), beta-caryophyllene (7.8%), beta-selinene (7.1%), and nothoapiole (5.8%) along with many other components in minor amounts. Seventeen components were identified in the oleoresin (Table 2) with dillapiole as a major component (30.7%). It also contains thymol (19.1%), nothoapiole (15.2.3%), and gamma-elemene (8.0%). The antioxidant activity of both the essential oil and oleoresin was evaluated in mustard oil by monitoring peroxide, thiobarbituric acid, and total carbonyl and p-anisidine values of the oil substrate. The results showed that both the essential oil and oleoresin were able to reduce the oxidation rate of the mustard oil in the accelerated condition at 60 degrees C in comparison with synthetic antioxidants such as butylated hydroxyanisole and butylated hydroxytoluene at 0.02%. In addition, individual antioxidant assays such as linoleic acid assay, DPPH scavenging activity, reducing power, hydroxyl radical scavenging, and chelating effects have been used. The C. nigrum seed essential oil exhibited complete inhibition against Bacillus cereus and Pseudomonas aeruginosa at 2000 and 3000 ppm, respectively, by agar well diffusion method. Antifungal activity was determined against a panel of foodborne fungi such as Aspergillus niger, Penicillium purpurogenum, Penicillium madriti, Acrophialophora fusispora, Penicillium viridicatum, and Aspergillus flavus. The fruit essential oil showed 100% mycelial zone inhibition against P. purpurogenum and A. fusispora at 3000 ppm in the poison food method. Hence, both oil and oleoresin could be used as an additive in food and pharmaceutical preparations after screening.     

Effects of dry period length on milk production, health, fertility, and quality of colostrum in dairy cows. Invited review

In terms of milk production, the dry period (DP) is a non-productive time. In addition, the risk of mastitis increases when cows with daily milk yield >12.5kg are dried off, which is often the case when the DP is set to 55-60 days. References on the positive effects of a shortened DP on the metabolic health at the start of lactation and the subsequent reproductive performance were the motive of this review. Completely foregoing or shortening the DP produces a decrease in milk yield during the following lactation. This decrease is particularly pronounced for cows at the beginning of the second lactation. It is thought that this may be due in part to different involution and proliferation processes of the mammary glands in primiparous and multiparous animals. The majority of the scientific papers suggest that shortening the DP has a positive rather than a negative effect on udder health. The quality of the colostrum is also unaffected; but if there is no drying off, the colostrum will be diluted. When considered overall, the literature, with support from metabolic profile studies, appears to favour a reduced impact on energy metabolism in early lactation in circumstances when the DP is shortened or waived. Clear positive effects on parameters relevant to energy metabolism however only apply to multiparous animals, and other factors, such as milking frequency before and after calving, exert an influence. Positive effects of shortening the DP on the reproductive performance were shown in several studies. Time from calving to first ovulation as well as indicators for assessing the success of artificial insemination and reproductive performance showed better results in animals with no or shortened drying off periods. Again, this only applied to multiparous animals. Overall, there is no single clear answer to the question of an optimal length of the DP, but conversely it is abundantly clear that a dry period of 55-60 days is not necessary in all cases and does not make sense under certain circumstances. Based on the study findings from recent years, the determination of dry periods should be used as a farm-specific and needs-oriented applied management decision.     

Fruit and seed characterization of wild populations of a traditional Andean crop: Solanum betaceum Cav. (Solanaceae) in the Argentinian Yungas

Genetic Resources and Crop Evolution - Chilto (Solanum betaceum Cav.) is a traditional Andean crop, appreciated for its high nutritional and commercial value. Despite it is cultivated in many...     

Development of antibodies for the detection of N-acetyl-glufosinate

Glufosinate is a widely used herbicide, which is difficult to detect by conventional analytical techniques. For many other herbicides, suitable antibodies have been raised for immunoassay development. Unfortunately, glufosinate is a very small molecule and difficult to immunize with. Thus, a derivatization-assisted immunoassay (DAIA) using the target analyte N-acetyl-glufosinate (NAG) was constructed. The activated hapten was synthesized by a new approach, using a homobifunctional cross-linker suberic acid bis(N-hydroxysuccinimide ester). The preparation of a suitable conjugate, the immunization, and the characterization of polyclonal antibodies are shown. The determination of the conjugation density (hapten density) of the immunogens was performed by four different methods (high-performance liquid chromatography with a refractive index detector, total reflection X-ray fluorescence, inductively coupled plasma mass spectrometry, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry), which gave similar results. The limit of detection was 17 mug/L NAG in water for the direct competitive enzyme immunoassay. NAG is also a main metabolite of glufosinate in resistant transgenic plants. The antibodies might be useful for the selective detection of NAG in the presence of the parent compound glufosinate (cross-reactivity 0.13%) and other metabolites.     

Evaluation of a TaqMan real-time PCR for the detection of Theileria parva in buffalo and cattle

A real-time PCR assay based on TaqMan probe chemistry was developed for the detection of Theileria parva DNA in blood samples. It uses a Theileria genus-specific PCR primer set and a T. parva-specific probe to amplify and hybridize with a species-specific part of the 18S rRNA gene of the parasite. The test was evaluated using positive and negative reference blood samples and shown to be specific for T. parva. Analytical sensitivity was determined by testing a dilution series of T. parva positive blood. It was shown to be able to detect parasitaemia as low as 2 × 10(-6)%. The Taqman assay results were also compared with that obtained with the real-time hybridization probe PCR assay, which is currently employed as the official test for the diagnosis of T. parva infections in buffalo and cattle and was shown to be equally sensitive. A panel of 1164 field samples was screened using both assays and 164 samples tested positive in both tests, indicating a good correlation.     

Length asymmetry of the bovine digits

The lengths of the digital bones of the fore- and hind-limbs obtained post mortem from 40 cattle of different ages were measured using digital radiographs. The lengths of the individual digital bones and the overall length of the digit were determined using computer software. The lateral metacarpal/metatarsal condyle, and lateral P1 and P2 were significantly longer than their medial counterparts, whereas P3 of the medial digit was longer than its lateral partner. Measured from the cannon bone epiphysis to the tip of the pedal bone, the mean increased length of the lateral digit was 0.8 mm in the fore- and 1.5 mm in the hind-limb. When the lengths of the digital bones were summed, the mean length of the lateral digit was 1.8 mm longer in the fore-limb and 2.1 mm longer in the hind-limb. Based on these findings, it can be concluded that the lengths of the paired digits differ in cattle. The majority of cattle have longer lateral digits in the fore- and hind-limbs. This asymmetry might explain why the lateral hind-limb claws are predisposed to sole ulcers on hard surfaces. In the hind-limbs, the impact is transferred from the pelvis directly to the longer lateral digit. In the fore-limb claws, the tenomuscular attachment to the trunk may be involved in a more even weight distribution and in a shift of weight to the medial claw.