Sesquiterpene Variation in West Australian Sandalwood (Santalum spicatum).

West Australian sandalwood (Santalum spicatum) has long been exploited for its fragrant, sesquiterpene-rich heartwood; however sandalwood fragrance qualities vary substantially, which is of interest to the sandalwood industry. We investigated metabolite profiles of trees from the arid northern and southeastern and semi-arid southwestern regions of West Australia for patterns in composition and co-occurrence of sesquiterpenes. Total sesquiterpene content was similar across the entire sample collection; however sesquiterpene composition was highly variable. Northern populations contained the highest levels of desirable fragrance compounds, α- and ß-santalol, as did individuals from the southwest. Southeastern populations were higher in E,E-farnesol, an undesired allergenic constituent, and low in santalols. These trees generally also contained higher levels of α-bisabolol. E,E-farnesol co-occurred with dendrolasin. Contrasting α-santalol and E,E-farnesol chemotypes revealed potential for future genetic tree improvement. Although chemical variation was evident both within and among regions, variation was generally lower within regions. Our results showed distinct patterns in chemical diversity of S. spicatum across its natural distribution, consistent with earlier investigations into sandalwood population genetics. These results are relevant for plantation tree improvement and conservation efforts.

Impacts of climate change and socio-economic scenarios on flow and water quality of the Ganges, Brahmaputra and Meghna (GBM) river systems: low flow and flood statistics.

The potential impacts of climate change and socio-economic change on flow and water quality in rivers worldwide is a key area of interest. The Ganges-Brahmaputra-Meghna (GBM) is one of the largest river basins in the world serving a population of over 650 million, and is of vital concern to India and Bangladesh as it provides fresh water for people, agriculture, industry, conservation and for the delta system downstream. This paper seeks to assess future changes in flow and water quality utilising a modelling approach as a means of assessment in a very complex system. The INCA-N model has been applied to the Ganges, Brahmaputra and Meghna river systems to simulate flow and water quality along the rivers under a range of future climate conditions. Three model realisations of the Met Office Hadley Centre global and regional climate models were selected from 17 perturbed model runs to evaluate a range of potential futures in climate. In addition, the models have also been evaluated using socio-economic scenarios, comprising (1) a business as usual future, (2) a more sustainable future, and (3) a less sustainable future. Model results for the 2050s and the 2090s indicate a significant increase in monsoon flows under the future climates, with enhanced flood potential. Low flows are predicted to fall with extended drought periods, which could have impacts on water and sediment supply, irrigated agriculture and saline intrusion. In contrast, the socio-economic changes had relatively little impact on flows, except under the low flow regimes where increased irrigation could further reduce water availability. However, should large scale water transfers upstream of Bangladesh be constructed, these have the potential to reduce flows and divert water away from the delta region depending on the volume and timing of the transfers. This could have significant implications for the delta in terms of saline intrusion, water supply, agriculture and maintaining crucial ecosystems such as the mangrove forests, with serious implications for people's livelihoods in the area. The socio-economic scenarios have a significant impact on water quality, altering nutrient fluxes being transported into the delta region.

Dynamic modeling of the Ganga river system: impacts of future climate and socio-economic change on flows and nitrogen fluxes in India and Bangladesh.

This study investigates the potential impacts of future climate and socio-economic change on the flow and nitrogen fluxes of the Ganga river system. This is the first basin scale water quality study for the Ganga considering climate change at 25 km resolution together with socio-economic scenarios. The revised dynamic, process-based INCA model was used to simulate hydrology and water quality within the complex multi-branched river basins. All climate realizations utilized in the study predict increases in temperature and rainfall by the 2050s with significant increase by the 2090s. These changes generate associated increases in monsoon flows and increased availability of water for groundwater recharge and irrigation, but also more frequent flooding. Decreased concentrations of nitrate and ammonia are expected due to increased dilution. Different future socio-economic scenarios were found to have a significant impact on water quality at the downstream end of the Ganga. A less sustainable future resulted in a deterioration of water quality due to the pressures from higher population growth, land use change, increased sewage treatment discharges, enhanced atmospheric nitrogen deposition, and water abstraction. However, water quality was found to improve under a more sustainable strategy as envisaged in the Ganga clean-up plan.

The transcriptome of sesquiterpenoid biosynthesis in heartwood xylem of Western Australian sandalwood (Santalum spicatum).

The fragrant heartwood oil of West Australian sandalwood (Santalum spicatum) contains a mixture of sesquiterpene olefins and alcohols, including variable levels of the valuable sesquiterpene alcohols, α- and ß-santalol, and often high levels of E,E-farnesol. Transcriptome analysis revealed sequences for a nearly complete set of genes of the sesquiterpenoid biosynthetic pathway in this commercially valuable sandalwood species. Transcriptome sequences were produced from heartwood xylem tissue of a farnesol-rich individual tree. From the assembly of 12,537 contigs, seven different terpene synthases (TPSs), several cytochromes P450, and allylic phosphatases were identified, as well as transcripts of the mevalonic acid and methylerythritol phosphate pathways. Five of the S. spicatum TPS sequences were previously unknown. The full-length cDNA of SspiTPS4 was cloned and the enzyme functionally characterized as a multi-product sesquisabinene B synthase, which complements previous characterization of santalene and bisabolol synthases in S. spicatum. While SspiTPS4 and previously cloned sandalwood TPSs do not explain the prevalence of E,E-farnesol in S. spicatum, the genes identified in this and previous work can form a basis for future studies on natural variation of sandalwood terpenoid oil profiles.

Control of eight predominant Eimeria spp. involved in economic coccidiosis of broiler chicken by a chemically characterized essential oil.

AIM: To control eight most predominant Eimeria spp. involved in the economic disease of coccidiosis in broiler chicken, by a chemically characterized essential oil of eucalyptus and peppermint. METHODS AND RESULTS: The experimental design consisted of 160 day-old-broiler chicks, divided into four equal groups (G1 , G2 , G3 and G4 ), with 40 birds per group. Each group was divided into four equal subgroups. Birds in G1 were deprived of essential oil treatment and of Eimeria challenge. Birds in G2 were unchallenged, and administered the essential oil in drinking water at 0.69 ml kg(-1) body weight. Birds in G3 were untreated with essential oil, and each of its four subgroups was challenged at a different age (14, 21, 28 and 35 days). Birds in G4 were treated with essential oil, and challenged in the same manner as for G3 . Equal number of birds from all subgroups (n = 10) were sacrificed at the sixth day after the time allocated for each challenge. The 6 day incubation period post challenge resulted in respective mean per cent weight increase in G2 and G1 birds equivalent to 57.8 and 53.1% (P < 0.05). In addition, the essential oil improved the per cent weight increase in challenged birds (54.6%) compared to the challenged-untreated birds (18.6%) (P < 0.05). The mean feed conversion, mortality, intestinal lesion scores and oocyst counts were significantly reduced in the challenged-treated birds compared to the challenged-untreated birds (P < 0.05). CONCLUSIONS: The results support the hypothesis of using the essential oils of eucalyptus and peppermint to control the most prevalent Eimeria spp. involved in coccidiosis of broiler chicken, helping in improvement of their production, alleviation of lesions and reduction in intestinal oocyst counts. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information about the possibility of using this blend of essential oil as a coccidiostat for the protection of broiler chickens against the prevalent eight Eimeria spp. of coccidiosis.

Modulation by essential oil of vaccine response and production improvement in chicken challenged with velogenic Newcastle disease virus.

AIM: To evaluate the impact of Eucalyptus and peppermint essential oils on immune modulation and production of broiler chicken challenged with a molecularly characterized velogenic NewCastle disease virus (vNDV). METHODS AND RESULTS: The experimental design included five treatments with three replicate pens/treatment comprised of 12-day-old broilers chicks/replicate. The five treatments included a positive challenge control (non-NDV vaccinated/nonessential oil treated/challenged) (NNEOC), a negative challenge control (NDV vaccinated/essential oil treated/unchallenged) (VEOU), a non-NDV vaccinated/essential oil treated/challenged (NEOC), a NDV vaccinated/nonessential oil treated/challenged (VNEOC) and a NDV vaccinated/essential oil treated/challenged (VEOC). The lowest mean survival rate (0·0%) and lowest production performance were obtained by the positive challenge control, while the best mean survival (93·3%) and average body weight (2649 g) were obtained by the negative challenge controls (P < 0·05). Among the three others challenged treatments, the best mean survival (79·2%), highest mean body weight at 42 days of age (2445 g), the lowest feed conversion ratio (1·60) and the highest serum conversion immunopotentiation at 35 days of age determined by ELISA and hemagglutination titres were obtained by the VEOC birds compared with respective means obtained by birds of the NEOC and VNEOC treatments (P < 0·05). CONCLUSIONS: The results supported the possibility of using the essential oils of Eucalyptus and Peppermint in broilers to immunopotentiate the response to vaccination against velogenic NDV, helping in significant improvement of survival and production. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information about the potential use of essential oils of eucalyptus and peppermint that can be exploited as commercial immunopotentiators for the protection of NDV-vaccinated broiler chickens against economic velogenic NDV.

Improving biological relevancy of transcriptional biomarkers experiments by applying the MIQE guidelines to pre-clinical and clinical trials.

The "Minimum Information for the Publication of qPCR Experiments" (MIQE [3]) guidelines are very much targeted at basic research experiments and have to our knowledge not been applied to qPCR assays carried out in the context of clinical trials. This report details the use of the MIQE qPCR app for iPhone (App Store, Apple) to assess the MIQE compliance of one clinical and five pre-clinical trials. This resulted in the need to include 14 modifications that make the guidelines more relevant for the assessment of this special type of application. We also discuss the need for flexibility, since while some parameters increase experimental quality, they also require more reagents and more time, which is not always feasible in a clinical setting.

Antimicrobial resistance of Staphylococcus species isolated from Lebanese dairy-based products.

The study evaluated the antimicrobial resistance of molecularly characterized strains of Staphylococcus aureus and S. saprophyticus isolated from 3 Lebanese dairy-based food products that are sometimes consumed raw: kishk, shanklish and baladi cheese. Suspected Staphylococcus isolates were identified initially using standard biochemical tests, then strains that were confirmed by polymerase chain reaction (29 S. aureus and 17 S. saprophyticus) were evaluated for their susceptibility to different antimicrobials. The highest levels of contamination with staphylococci were in baladi cheese. Resistance rates ranged from 67% to gentamicin to 94% to oxacillin and clindamycin. The results suggest that these locally made dairy-based foods may act as vehicles for the transmission of antimicrobial-resistant Staphylococcus spp.

Impact of sex, age, race, ethnicity and aspirin use on bleeding symptoms in healthy adults.

BACKGROUND: Comparing a patient's bleeding symptoms with those of healthy individuals is an important component of the diagnosis of bleeding disorders, but little is known about whether bleeding symptoms in healthy individuals vary by sex, race, ethnicity, age, or aspirin use. OBJECTIVES, PATIENTS/METHODS: We developed a comprehensive, ontology-backed, Web-based questionnaire to collect bleeding histories from 500 healthy adults. The mean age was 43 years (range 19-86 years), 63% were female, 19% were Hispanic, 37% were African-American, 43% were Caucasian, 8% were Asian, and 4% were multiracial. RESULTS: 18 of the 36 symptoms captured occurred with < 5% frequency, and 26% of participants reported no bleeding symptoms (range 0-19 symptoms). Differences in sex, race, ethnicity, aspirin use and age accounted for only 6-13% of the variability in symptoms. Although men reported fewer symptoms than women (median 1 vs. 2, P < 0.01), there was no difference when sex-specific questions were excluded (median 1 for both men and women, P = 0.50). However, women reported more easy bruising (24% vs. 7%, P < 0.01) and venipuncture-related bruising (10% vs. 3%, P = 0.02). The number of symptoms did not vary by race or age, but epistaxis was reported more frequently by Caucasians than by African-Americans (29% vs. 18%, P = 0.02), and epistaxis frequency decreased with age (odds ratio 0.97 per year, P < 0.01). Paradoxically, infrequent aspirin users reported more bruising and heavy menses than frequent users (21% vs. 8%, P = 0.01, and 56% vs. 38%, P = 0.03, respectively). CONCLUSIONS: Our findings provide a contemporaneous and comprehensive description of bleeding symptoms in a diverse group of healthy individuals. Our Web-based system is freely available to other investigators.

Antimicrobial resistance of Staphylococcus species isolated from Lebanese dairy-based products

The study evaluated the antimicrobial resistance of molecularly characterized strains of Staphylococcus oureus and S. saprophyticus isolated from 3 Lebanese dairy-based food products that are sometimes consumed raw: kishk, shanklish and baladi cheese. Suspected Staphylococcus isolates were identified initially using standard biochemical tests, then strains that were confirmed by polymerase chain reaction [29 S, aureus and 17 S. saprophyticus] were evaluated for their susceptibility to different antimicrobials. The highest levels of contamination with staphylococci were in baladi cheese. Resistance rates ranged from 67% to gentamicin to 94% to oxacillin and clindamycin. The results suggest that these locally made dairy-based foods may act as vehicles for the transmission of antimicrobial-resistant Staphylococcus spp

Antimicrobial resistance and pathogenicity of Escherichia coli isolated from common dairy products in the Lebanon.

In a recent study, bacteria have been isolated from popular Lebanese dairy products, which had been collected in the Beqaa Valley, in north-eastern Lebanon. The foods investigated were two cheeses (shankleesh and baladi) and a dried fermented mixture of yogurt and wheat grains (kishk). Bacterial colonies on McConkey and sorbitol-McConkey agar that showed the morphology of Escherichia coli were biochemically tested and then classified, using PCR-based assays, into the various strains of pathogenic and non-pathogenic E. coli. Some of the confirmed E. coli isolates were proven to be pathogenic, including two identified as E. coli O157:H7. When the pathogenic isolates were tested for their susceptibility to 10 different antibiotics (all commonly used, by clinicians and veterinarians, for the treatment of infections with Gram-negative bacteria), each tested isolate was found to be highly resistant to at least one antibiotic. It therefore appears that, in Lebanon, some popular dairy products pose a public-health hazard, acting as vehicles for the transmission of drug-resistant pathogens.

Anti-inflammatory bioactivities in plant extracts.

The medical ethnobotanical knowledge propagated over generations in the coastal regions of the Eastern Mediterranean, including Lebanon, is one that has built on several ancient cultures and civilizations of these regions. Recent interest in medical ethnobotany and the use of medicinal herbs in treating or preventing ailments has rejuvenated interest in folk medicine practices, especially those transcendent across generations. According to Eastern Mediterranean folk medicine practices, herbal remedies that treat many inflammation-related ailments were typically based on plant bioactive water extracts or decoctions. Studies have shown that active anti-inflammatory ingredients in water extracts include many natural chemicals such as phenols, alkaloids, glycosides, and carbohydrates. The intent of this manuscript is twofold: first, to review the literature that describes anti-inflammatory bioactivities in plant extracts of different plant genera; and second, to evaluate indigenous folk remedies used by folk doctors to treat inflammatory ailments in this region of the world. For this aim, the reported literature of five plant genera assumed to possess anti-inflammatory bioactivities and typically prescribed by folk doctors to treat inflammation-related ailments is reviewed.

Competitive exclusion against Salmonella Enteritidis in layer chickens by yoghurt microbiota: impact on egg production, protection and yolk-antibody and cholesterol levels.

AIMS: This work aims at studying the impact of competitive exclusion of Salmonella serotype Enteritidis infection in layer chickens, by microbiota of fresh and dried-modified yoghurt, on egg production and weight, protection against infection, and on yolk-antibody and cholesterol levels. METHODS AND RESULTS: Four groups of 27-week-old layer chickens were included in this study. After an initial enrofloxacin treatment, groups 1 and 2 were administered fresh or dry yoghurt (respectively) for 14 days. Groups 1, 2 and 3 were challenged intraoesophageally with Salm. Enteritidis, on the sixth day of yoghurt administration, while group 4 was left unchallenged and without yoghurt administration. No significant difference in percent infectivity of visceral organs with Salm. Enteritidis was observed between the groups. The yoghurt administered groups showed an early significant antibody response in their yolk on the seventh day postchallenge (P < 0.05) and highest egg production and weight. Finally, the egg yolk cholesterol concentration was higher in Salm. Enteritidis-challenged groups than that observed in the unchallenged group. CONCLUSIONS: The results point to the possible involvement of yoghurt administration in immunopotentiation and improvement of egg production and weight. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings warrant further research that could improve immunity and production in layer chickens infected with Salm. Enteritidis.

Microbiological and chemical properties of litter from different chicken types and production systems.

Chicken litter is produced in large quantities from all types of poultry raising activities. It is primarily used for land application, thus it is essential to analyze its properties before it is released to the environment. The objective of this study is to compare the microbiological and chemical properties of litter generated from layer and broiler chickens reared under intensive and free-range production systems. The microbiological analysis consisted of the enumeration of total bacteria, total coliforms, Staphylococcus species, Salmonella species and Clostridium perfringens. Chicken litter from layers reared under intensive and free range systems showed lower mean total bacterial count than the litter collected from chicken broilers reared under either of the two systems (P=0.0291). The litter from intensive layers had the lowest mean total coliform counts (P=0.0222) while the lowest Staphylococcus species count was observed in the litter from free-range layers (P=0.0077). The C. perfringens count was the lowest in chicken litter from intensively raised broilers and layers (P=0.0001). The chemical properties of litter from the different chicken types and production systems were compared based on determination of pH, electrical conductivity, carbon, nitrogen, phosphorus, potassium, cadmium and zinc. Litter from free-range broilers showed the highest pH value (P=0.0005); however, the electrical conductivity was higher in the litter from both intensive and free-range layers compared to the litter from both broiler production systems (P=0.0117). Chicken litter from intensive systems had higher nitrogen content than litter from free-range systems (P=0.0000). The total phosphorus was the lowest in free-range broiler litter (P=0.0001), while the total potassium was the lowest in litter from intensively managed broilers (P=0.0000). Zinc appeared higher in litter from layers compared to that from broilers (P=0.0101). The cadmium content was higher in the litter from free-range broilers and layers compared to that in the litter from intensively managed systems (P=0.0439). Staphylococcus species in the litter as well as cadmium concentrations seem to be the most critical parameters presenting risks on the environment and on human health. Based on the lowest coliform counts (an indication of water pollution), the high nutrient levels and the low cadmium values, litter from intensively managed layers appears as the most suitable for application on agricultural soils.

Avian influenza outbreak in poultry in the Lebanon and transmission to neighbouring farmers and swine.

Twenty-four poultry farms in four major provinces of the Lebanon were investigated to verify the first emergence of avian influenza (AI). Both the meat chicken breeders and commercial chicken layers presented a significant average drop in egg production equivalent to 46% and 47.3%, respectively. However, the average drop in egg production in the free-range layers was only 11.1%. Flocks were confirmed as positive for AI by ELISA, clinical signs and pathological lesions. The pathogenicity, including case fatality in affected chickens, was different depending of the types of chicken and farming conditions. The average fatality rates among breeders, commercial layers, free-range layers and broilers were 2%, 2%, 1% and 35%, respectively. The majority of the randomly selected ELISA-positive serum samples collected from different farms showed H9-specific haemagglutination inhibition (HI) antibodies. The direct immunoflorescent antibody test also revealed the presence of H9 antigen in congested brains and in tracheal lesions of broilers. The virus isolated from the brains of broilers was subtype H9N2. Pigs fed with carcasses of affected chickens showed H9-specific HI antibodies at 100%. Farmers (32.3%) serving the affected chickens also revealed these antibodies in their sera.

Comparison of immunosuppression in dry and lactating Awassi ewes due to water deprivation stress.

In seminomadic farming practice, dry and lactating ewes are exposed to different degrees of water deprivation, leading to stress followed by various disease outbreaks. This study compares quantitatively the immunosuppression to Salmonella Enteritidis (SE) fimbriae (14 and 21 kDa) and other major polypeptides (28.9, 37.7, 42.9, 68.0, 92.6 and 96.8 kDa) in water-deprived dry and lactating ewes. Sixteen dry and lactating multiparous Awassi ewes were divided into four treatment groups (A, A', B and B'). Ewes in groups A and B were lactating, whereas ewes in groups A' and B' were dry. All ewes were administered a killed SE vaccine, subcutaneously in the neck, at the initiation of the experiment. The water availability for ewes in groups B (lactating) and B' (dry) was ad libitum, while that for ewes in groups A (lactating) and A' (dry) was once every 4 days. A serum sample was collected from the jugular vein of each ewe at zero time (initiation of the experiment, when SE bacterin was delivered) and at 2, 9, 12, 15 and 18 days post SE vaccination. The percentage reduction in the level of humoral antibody response to polypeptides of > or = 21 kDa was more apparent in water-deprived lactating ewes of group A between 9 and 18 days post initiation of thirst. In this period, immunosuppression to polypeptides > or = 21 kDa was present in 14 out of 16 observations in group A (water-deprived lactating), with significant immunosuppression in 9 observations in relation to the respective control (p<0.05), while it was present in only 4 out of 16 observations in group A' (water-deprived dry), with significant immunosuppression in 2 observations (p <0.05). In conclusion, immunosuppression to polypeptides of > or =21 kDa is more significant in lactating water-deprived ewes in the period 9-18 days post initiation of thirst, a result that will influence our future sheep welfare awareness programmes targeting an elimination of the practice of water deprivation in seminomadic sheep farming.

Comparative impact of live chicken infectious anaemia virus vaccine versus natural exposure in meat chicken breeders on immunity to infectivity by CIA and inclusion body hepatitis viruses in their offspring.

The immunology and histopathology and the distribution of viral antigen in infections with chicken infectious anaemia virus (CIAV) and inclusion body hepatitis virus (IBHV) were compared in the broiler offspring of CIAV-vaccinated meat chicken breeders versus those in the offspring of breeders naturally exposed to field CIAV. No significant difference in the humoral antibody level specific for CIAV was observed between 5 and 33 weeks of age in the two breeder groups (p > 0.05). The maternal humoral immunity to CIAV in the day-old offspring of the groups did not differ significantly (p > 0.05). The humoral immunity to CIAV at 40 days of age indicated an absence of clinical signs of CIAV in the broiler offspring of both groups of breeders and this was associated with mean serum thymulin levels in offspring of both groups not differing significantly at 1 or 40 days of age. Histopathological and immunofluorescence observations did not differ significantly in the offspring of either group by CIAV or IBHV.

Evaluation of 12 stabilizers in a developed attenuated Salmonella Enteritidis vaccine.

The development of a stable live attenuated Salmonella Enteritidis (SE) vaccine, resisting heat stress during transportation and storage in unequipped tropical and subtropical zones of the world, is highly recommended. Twelve stabilizers were individually supplemented into a 9 ml volume of sterile distilled water resulting in concentrations of 1, 2, 3, 4 and 5%. A volume of 1 ml of attenuated live SE vaccine is added over the 9 ml of each concentration of the stabilizers. The differently stabilized SE vaccines were stressed at 55 degrees C for 48 h. The lowest percent reductions in SE cell viability by specified level of each stabilizer in ascending order were: 22.3% by 2% skim milk, 55.1% by 5% bovine serum albumin (BSA), 59.2% by 4% sorbitol, 74.4% by 3% maltose, 75% by 2% honey, 91.3% by 3% histidine, 96.9% by 1% heparin, 97.5% by 4% dextrose, 97.9% by 5% lactose, 99.4% by 5% sucrose, 99.5% by 2% gelatin, and 100% by 1-5% glycerol. In narrowing the concentration levels of skim milk to include 1.00, 1.25, 1.50, 1.75, 2.00, 2.25, 2.50, 2.75, and 3.00%, the 2.50% was the optimum level resulting in minimal percent reduction in SE cell viability of 18.9% after exposure to the defined heat stress.