Maritime Halophyte Species from Southern Portugal as Sources of Bioactive Molecules

Extracts of five halophytes from southern Portugal (Arthrocnemum macrostachyum, Mesembryanthemum edule, Juncus acutus, Plantago coronopus and Halimione portulacoides), were studied for antioxidant, anti-inflammatory and in vitro antitumor properties. The most active extracts towards the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical were the methanol extracts of M. edule (IC₅₀ = 0.1 mg/mL) and J. acutus (IC₅₀ = 0.4 mg/mL), and the ether extracts of J. acutus (IC₅₀ = 0.2 mg/mL) and A. macrostachyum (IC₅₀ = 0.3 mg/mL). The highest radical scavenging activity (RSA) against the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical was obtained in the ether extract of J. acutus (IC₅₀ = 0.4 mg/mL) and H. portulacoides (IC₅₀ = 0.9 mg/mL). The maximum total phenolic content (TPC) was found in the methanol extract of M. edule (147 mg gallic acid equivalents (GAE)/g) and in the ether extract of J. acutus (94 mg GAE/g). Significant decreases in nitric oxide (NO) production were observed after incubation of macrophages with lipopolysaccharide (LPS) and the chloroform extract of H. portulacoides (IC₅₀ = 109 µg/mL) and the hexane extract of P. coronopus (IC₅₀ = 98.0 µg/mL). High in vitro cytotoxic activity and selectivity was obtained with the ether extract of J. acutus. Juncunol was identified as the active compound and for the first time was shown to display selective in vitro cytotoxicity towards various human cancer cells.     

Dose determination studies for monepantel, an amino-acetonitrile derivative, against fourth stage gastro-intestinal nematode larvae infecting sheep

Monepantel is the first compound from the recently discovered amino-acetonitrile derivative (AAD) class of anthelmintics to be developed for use in sheep. Three dose determination studies were conducted in Australia and Switzerland to identify the minimum therapeutic dose of monepantel when formulated for the oral treatment of sheep to control fourth stage (L4) gastro-intestinal nematode larvae. In each study, sheep infected with the target nematodes (selected from Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta, Teladorsagia trifurcata, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Cooperia curticei, Cooperia oncophora, Nematodirus battus, Nematodirus filicollis, Nematodirus spathiger, Chabertia ovina and Oesophagostomum venulosum) were treated with either 1.25, 2.5 or 5.0mg monepantel/kg liveweight. Following euthanasia and worm counting, efficacy was calculated against worm counts from untreated control groups. Monepantel proved highly effective at 2.5 and 5.0mg/kg, but was only moderately effective against some nematode species (L4 stage) at 1.25mg/kg. The results also confirmed that monepantel will effectively control L4 stages of nematodes resistant to at least some of the currently available broad-spectrum anthelmintic classes (macrocyclic lactone resistant strains were not included in the studies). It was concluded that 2.5mg/kg would be a suitable minimum dose rate for a commercial product. No adverse events related to treatment with monepantel were detected.     

Effects of supplemental d-methionine in comparison to l-methionine on nitrogen retention,gut morphology,antioxidant status,and mRNA abundance of amino acid transporters in weanling pigs

An N-balance experiment was conducted to test the hypothesis that d-Methionine (d-Met) has the same bioavailability and efficacy as l-Methionine (l-Met) when fed to weanling pigs. A Met-deficient basal diet containing 0.24% standardized ileal digestible (SID) Met was formulated. Six additional diets were formulated by adding 0.036%, 0.072%, or 0.108% d-Met or l-Met to the basal diet, and these diets, therefore, contained 77%, 87%, or 97% of the requirement for SID Met. Fifty-six barrows (10.53 ± 1.17 kg) were housed in metabolism crates and allotted to the seven diets with eight replicate pigs per diet. Feces and urine were collected quantitatively with 7-d adaptation and 5-d collection periods. Blood and tissue samples from pigs fed the basal diet and pigs fed diets containing 0.108% supplemental Met were collected on the last day. Results indicated that N retention (%) linearly increased (P < 0.01) as supplemental d-Met or l-Met increased in diets. Based on N retention (%) as a response, the linear slope-ratio regression estimated the bioavailability of d-Met relative to l-Met to be 101% (95% confidence interval: 57%–146%). The villus height and crypt depth in the jejunum were not affected by the Met level or Met source. Total antioxidant capacity or thiobarbituric acid reactive substance concentrations in plasma or tissue samples from pigs fed the control diet or diets containing 0.108% supplemental d-Met or l-Met were not different. Abundance of mRNA for some AA transporters analyzed in intestinal mucosa of pigs also did not differ. Therefore, it is concluded that d-Met and l-Met are equally bioavailable for weanling pigs.     

Growth performance and carcass quality are not different between pigs fed diets containing cold-fermented low-oil DDGS and pigs fed conventional DDGS,but pelleting improves gain to feed ratio regardless of source of DDGS

The objective of this study was to test the hypothesis that growth performance and carcass characteristics of pigs fed diets containing cold-fermented, low oil distillers dried grains with solubles (DDGS) is not different from that of pigs fed diets containing conventional DDGS regardless of the physical form of the diets. A total of 160 barrows and gilts were used. There were 4 diets, 10 pens per diet, and 4 pigs per pen. Pigs were weaned at 21 d of age and fed a common phase 1 diet that did not contain DDGS during the initial 7 d post-weaning. Pigs were then allotted to the four diets that were arranged in a 2 × 2 factorial design with two sources of DDGS (cold-fermented and conventional DDGS) and two diet forms (meal and pellets). Pigs were fed phase 2 diets from day 7 to 21 and phase 3 diets from day 21 to 43 post-weaning. All diets were based on corn and soybean meal, but phase 2 diets also contained 15% DDGS and phase 3 diets contained 30% DDGS. From day 43, pigs were fed grower diets for 38 d, early finisher diets for 38 d, and late finisher diets for 18 d and these diets also contained 30% DDGS. Feed was provided on an ad libitum basis and daily feed allotments were recorded. Pigs were weighed at the beginning of each phase and at the conclusion of the experiment. On the last day of the experiment, the pig in each pen with a body weight that was closest to the pen average was slaughtered and carcass measurements were determined. Combined results for the two nursery phases indicated that feeding meal diets instead of pelleted diets increased (P < 0.001) average daily feed intake and decreased (P < 0.05) gain to feed ratio (G:F). However, no differences between the two sources of DDGS were observed for the overall growth performance of weanling pigs. For the entire growing-finishing period, the source of DDGS did not affect growth performance, but pigs fed meal diets had reduced (P < 0.001) G:F compared with pigs fed the pelleted diets. There were no differences between the two sources of DDGS for carcass characteristics. Back fat was greater (P < 0.05) for pigs fed pelleted diets than for pigs fed meal diets. In conclusion, no differences in growth performance or carcass characteristics between pigs fed cold-fermented DDGS and pigs fed conventional DDGS were observed. However, pigs fed pelleted diets had greater G:F and greater back fat than pigs fed meal diets.     

Livestock Browsing,Not Water Limitations,Contributes to Recruitment Failure of Dobera glabra in Semiarid Ethiopia

The study used nursery and field experiments to investigate why recruitment of Dobera glabra (Forssk) Poir., a native food source for both humans and livestock, often fails in the semiarid rangelands of Afar, Ethiopia. We hypothesized that soil water limitations and browsing by livestock would be the primary mechanisms accounting for the failure of natural regeneration. We used three sets of experiments—1) seedling performance in response to shade and watering in a nursery, 2) field regeneration with and without browsing, and 3) regeneration beneath trees with and without browsing—to examine regeneration success. Seedlings were established in plots from seeds sown directly into the soil for nursery and field experiments, but natural germination occurred beneath trees. Survival and relative growth rates (RGR) were used to monitor seedling performance. Seedlings that received neither shade nor watering treatments had lower seedling survival (53 ± 15%) as compared with other treatment combinations. Highest seedling survival was recorded under shade and 1 d watering s wk^-1 treatment combinations (92 ± 1%). However, shade treatments had a minimal increase on seedling RGR. Water limitation is not a crucial limiting factor for D. glabra recruitment, as 53% of the seedlings survived without both shade and supplemental water for 1 yr. Field experiments, however, demonstrated that browsing greatly reduced seedling survival (below 15%) and suppressed growth of surviving seedlings, suggesting that browsing is the major factor preventing natural recruitment. Natural recruitment of D. glabra is unlikely with the existing continuous and intensive grazing/browsing in Afar rangelands, where the mobility of pastoralists is restricted. We suggest that planting nursery-raised seedlings in home gardens of settled pastoralists and establishment of grazing reserves in some key range sites that contain D. glabra could help offset the recruitment failure of native food species D. glabra in Afar rangelands.     

Barbiturate intoxication in two dogs confirmed by toxicological urinalysis

Two dogs were presented within 24 hours to the Department of Small Animal Medicine and Surgery at the University of Veterinary Medicine Hannover for investigation of the sudden onset of neurological abnormalities following a walk in the same park. One dog was observed ingesting a piece of meat. Analysis of urine by gas chromatography-mass spectrometry from each of the dogs identified the presence of barbiturates. Both dogs recovered with supportive treatment. This is the first report to describe the use of toxicological urinalysis with gas chromatography-mass spectrometry for the diagnosis of barbiturate intoxication in dogs.     

A review of prevalences of paratuberculosis in farmed animals in Europe

Prevalence estimates are used by decision makers such as policy makers and risk assessors to make choices related to certain diseases and infections. Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic infection particularly resulting in economic losses among farmed ruminants. Therefore, this infection is of concern for many farming industry decision makers. As a result, multiple studies have been carried out to determine the within-herd and between-herd prevalence of MAP infections. The objective of the present study was to conduct a review and, if possible, compare animal and herd level prevalences of MAP among farmed animals in Europe. European data on prevalences of MAP in all farmed animal species were included from a review of literature. Information on target population and study design, tests used and apparent prevalences were recorded, and subsequently true prevalences were calculated when possible. A critical review of the included studies indicated that although a wide range of studies have been conducted, likely and comparable true prevalence estimates could rarely be calculated. Based on a few studies where the prevalences appeared to be plausible, it was concluded that prevalences of MAP would have to be guesstimates based on available data. The true prevalence among cattle appeared to be approximately 20% and was at least 3-5% in several countries. Between-herd prevalence guesstimates appeared to be >50%. No countries had published sufficient information to claim freedom from MAP or just a near-zero prevalence of MAP infections. No within-flock prevalence estimates were available for goats and sheep. The between-flock prevalence guesstimates were >20%, based only on estimates from Switzerland and Spain. Major critical issues were identified in the majority of the studies, primarily due to lack of knowledge of the test accuracy of the diagnostic test used, or due to studies where the study population did not reflect the target population. Because most of the reported studies did not yield prevalence estimates which were comparable to other studies, there is a continuing need for well-designed studies of the prevalence of MAP infections.     

Identification of fatty acid esters of pectenotoxin-2 seco acid in blue mussels (Mytilus edulis) from Ireland

Pectenotoxins from marine dinoflagellates of the genus Dinophysis are rapidly hydrolyzed by many shellfish to give pectenotoxin-2 seco acid, which isomerizes to 7-epi-pectenotoxin-2 seco acid. Three series of fatty acid esters of pectenotoxin-2 seco acid (PTX-2 seco acid) and 7-epi-PTX-2 seco acid were detected by LC-MS analysis of extracts from blue mussels (Mytilus edulis) from Ireland. The locations of the fatty acid ester linkages were identified by a combination of LC-MSn in positive- and negative-ion modes, LC-MS analysis of the products from reaction of the esters with sodium periodate, and NMR analysis of purified samples of the two most abundant ester derivatives. The 37-O-acyl esters of PTX-2 seco acid were the most abundant, followed by the corresponding 11-O-acyl esters, accompanied by low levels of the 33-O-acyl esters. The most abundant fatty acid esters in the fractionated sample were, in order, the 16:0, 22:6, 14:0, 16:1, 18:4, and 20:5 fatty acids, although a wide array of other PTX-2 seco acid fatty acid esters were also present at low levels.