Detection and quantification of equine herpesvirus-1 viremia and nasal shedding by real-time polymerase chain reaction.

Equine herpesvirus-1 (EHV-1) infection is common in young horses throughout the world, resulting in respiratory disease, epidemic abortion, sporadic myelitis, or latent infections. To improve on conventional diagnostic tests for EHV-1, a real-time polymerase chain reaction (PCR) technique was developed, using primers and probes specific for the EHV-1 gB gene. Amplification efficiencies of 100% +/- 5% were obtained for DNA isolated from a plasmid, infected peripheral blood mononuclear cells (PBMCs), and nasal secretions from infected ponies. The dynamic range of the assay was 8 log10 dilutions, and the lower limit of detection was 6 DNA copies. Fifteen ponies, seronegative for EHV-1, were experimentally infected with EHV-1, and nasal samples were used to quantify shedding of virus by both virus isolation and real-time PCR analysis. Virus isolation identified nasal shedding of EHV-1 in 12/15 ponies on a total of 25 days; real-time PCR detected viral shedding in 15/15 ponies on 75 days. Viremia was quantified using PBMC DNA, subsequent to challenge infection in 3 additional ponies. Viremia was identified in 1/3 ponies on a single day by virus isolation; real-time PCR detected viremia in 3/3 ponies on 17 days. When real-time PCR was used to analyze PBMC DNA from 11 latently infected ponies (documented by nested PCR), EHV-1 was not detected. We conclude that real-time PCR is a sensitive and quantitative test for EHV-1 nasal shedding and viremia and provides a valuable tool for EHV-1 surveillance, diagnosis of clinical disease, and investigation of vaccine efficacy.     

Anthropogenic influences on potential fire spread in a pyrogenic ecosystem of Florida, USA

Fire has historically been an important ecological factor maintaining southeastern U.S. vegetation. Humans have altered natural fire regimes by fragmenting fuels, introducing exotic species, and suppressing fires. Little is known about how these alterations specifically affect spatial fire extent and pattern. We applied historic (1920 and 1943) and current (1990) GIS fuels maps and the FARSITE fire spread model to quantify the differences between historic and current fire spread distributions. We held all fire modeling variables (wind speed and direction, cloud cover, precipitation, humidity, air temperature, fuel moistures, ignition source and location) constant with exception of the fuel models representing different time periods. Model simulations suggest that fires during the early 1900's burned freely across the landscape, while current fires are much smaller, restricted by anthropogenic influences. Fire extent declined linearly with patch density, and there was a quadratic relationship between fire extent and percent landscape covered by anthropogenic features. We found that as little as 10 percent anthropogenic landcover caused a 50 percent decline in fire extent. Most landscapes (conservation or non-conservation areas) are now influenced by anthropogenic features which disrupt spatial fire behavior disproportionately to their actual size. These results suggest that land managers using fire to restore or maintain natural ecosystem function in pyrogenic systems will have to compensate for anthropogenic influences in their burn planning. This revised version was published online in May 2005 with corrections to the Cover Date. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of Artificial Diets on Growth,Survival and Condition of Adult Cuttlefish, <Emphasis Type="Italic">Sepia officinalis</Emphasis> Linnaeus, 1758

The effects of artificial diets on growth and body condition of adult cuttlefish, Sepia officinalis were tested in two experiments. Supplemented prepared diets (fish myofibrillar protein concentrate) were fed during a 30-day and a 21-day experiments. Growth, feeding rate and food conversion of group-reared cuttlefish were analyzed. The first of these experiments tested four artificial diets, made with increasing levels of lysine, on adult cuttlefish. According to the chemical analysis, diets 1–3 had limiting concentrations of lysine and other essential amino acids (compared to mantle composition of the cuttlefish), while diet 4 was the only one where almost all essential amino acids were present in concentrations similar or higher than the ones present in cuttlefish mantle. A second experiment was conducted by isolating 16 adult cuttlefish individually, and feeding them the same four artificial diets, in order to obtain individual data. During Experiment 1, only the diet with the best chemical score (diet 4) produced growth (p < 0.05), with a mean instantaneous growth rate (MIGR) of 0.30% wet body weight (BW) d⁻¹. Similarly, individually reared cuttlefish fed diet 4 produced the highest IGR’s (0.26, 0.38 and 0.48% BW d⁻¹) and grew larger (p < 0.01). Comparison of cuttlefish fed the artificial diets vs. thawed shrimp and unfed cuttlefish indicated that cuttlefish fed the artificial diets were in an intermediate state. Growth rates obtained with the artificial diets (<0.4% BW d⁻¹) were considerably lower compared to natural prey, live or frozen, reported by other authors.     

Impact of foot-and-mouth disease on mastitis and culling on a large-scale dairy farm in Kenya

Foot and mouth disease (FMD) is a highly transmissible viral infection of cloven hooved animals associated with severe economic losses when introduced into FMD-free countries. Information on the impact of the disease in FMDV-endemic countries is poorly characterised yet essential for the prioritisation of scarce resources for disease control programmes. A FMD (virus serotype SAT2) outbreak on a large-scale dairy farm in Nakuru County, Kenya provided an opportunity to evaluate the impact of FMD on clinical mastitis and culling rate. A cohort approach followed animals over a 12-month period after the commencement of the outbreak. For culling, all animals were included; for mastitis, those over 18 months of age. FMD was recorded in 400/644 cattle over a 29-day period. During the follow-up period 76 animals were culled or died whilst in the over 18 month old cohort 63 developed clinical mastitis. Hazard ratios (HR) were generated using Cox regression accounting for non-proportional hazards by inclusion of time-varying effects. Univariable analysis showed FMD cases were culled sooner but there was no effect on clinical mastitis. After adjusting for possible confounders and inclusion of time-varying effects there was weak evidence to support an effect of FMD on culling (HR = 1.7, 95% confidence intervals [CI] 0.88-3.1, P = 0.12). For mastitis, there was stronger evidence of an increased rate in the first month after the onset of the outbreak (HR = 2.9, 95%CI 0.97-8.9, P = 0.057).

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-015-0173-4) contains supplementary material, which is available to authorized users.     

Effect of Fermentation and Oil Incorporation on the Retention of E Vitamers During Breadmaking

All forms of baking and processing cause a loss of nutrients, including vitamin E, but little is known about these occurrences or if they could be avoided. The objective of this research was to study the incorporation of palm oil and the stability of vitamin E in palm oil during breadmaking. Wheat meal and rye breads were baked with and without the addition of 0, 2, 5, or 8% palm oil. The eight E group vitamers (tocopherols and tocotrienols) were extracted by using accelerated solvent extraction, freeze dried, and then analyzed with normal‐phase HPLC. Compared with the controls, the inclusion of palm oil was found to increase the quantity of all forms of vitamin E in the final baked products. It is concluded that palm oil is effective in increasing the vitamin E content of whole grain bread.     

Introgression of antioxidant activity into cassava (Manihot esculenta C.): an effective technique for extending fresh storage roots shelf life

Physiological postharvest deterioration (PPD) of cassava is the main constraint affecting its nutritional and economical values. PPD is induced by wounds when detaching storage roots from mother plant during harvesting. It is accelerated by the reactive oxygen species (ROS) such as oxygen ion (O₂) and peroxide (O₂)^?2. The carotenoid content and its antioxidant property can help in extending shelf life of cassava storage roots. The primary mode of action of carotenoids as antioxidant is to quench singlet oxygen. Cassava breeding was reported to successfully introgress and improve carotenoid content into cassava. The two types of phytoene synthase (PSY) enzymes (PSY1 and PSY2) are key regulators of carotenoids accumulation in cassava. Carotenoids formation and accumulation in cassava storage roots are induced by a single nucleotide polymorphism in PSY2 which causes a non‐conservative amino acid exchange. This single nucleotide polymorphism in PSY gene is co‐segregated with β‐carotene in cassava storage roots, a phenomenon that could help to unravel the mechanism of introgression of carotenoids into cassava. This article investigates breeding feasibility for improving quality of cassava landraces in developing countries.     

Gas chromatography for detection of citrus infestation by fruit fly larvae (Diptera: Tephritidae)

Tephritid fruit flies are serious economic pests worldwide. As larvae, they feed and develop within the pulp of host fruits, making infestation difficult to detect by visual inspection. At U.S. ports of entry, incoming produce shipments are checked for infestation by manually cutting open a small sample of fruit and searching for tephritid larvae. Consequently, there is a need for more sensitive, high-throughput screening methods. This study evaluated gas chromatography (GC) as a potential technology for improved detection of hidden infestation. Grapefruits (Citrus × paradisi Macfad.) infested with immature stages of the Caribbean fruit fly Anastrepha suspensa (Loew) (Diptera: Tephritidae) were examined to determine if infested fruit emitted a chemical profile distinct from that of non-infested fruit. Peaks identified by GC analysis were grouped into three classes. Chemicals detected in similar quantities in all samples, or slightly elevated in infested samples, were regarded as non-diagnostic background volatiles. Chemicals highly elevated after oviposition, during the last instar exit stage, and in experimentally-pierced fruit were interpreted to be indicators of citrus peel injury, and included d-limonene and β-ocimene. Chemicals elevated exclusively in the larval infestation stages were considered indicators of feeding damage and potentially diagnostic of infestation, and included hexyl butanoate and an unidentified compound. The peaks associated with injury and feeding were also detectable with a portable ultra-fast GC analyzer that required less than 80 s per sample. Further studies will investigate the potential application of these results for development of a rapid, non-destructive screening method for detection of tephritid infestation.     

Crystal structure of the long-chain fatty acid transporter FadL

The mechanisms by which hydrophobic molecules, such as long-chain fatty acids, enter cells are poorly understood. In Gram-negative bacteria, the lipopolysaccharide layer in the outer membrane is an efficient barrier for fatty acids and aromatic hydrocarbons destined for biodegradation. We report crystal structures of the long-chain fatty acid transporter FadL from Escherichia coli at 2.6 and 2.8 angstrom resolution. FadL forms a 14-stranded beta barrel that is occluded by a central hatch domain. The structures suggest that hydrophobic compounds bind to multiple sites in FadL and use a transport mechanism that involves spontaneous conformational changes in the hatch.