Development of an enzyme-linked immunosorbent assay for the fungicide imazalil in citrus fruits

Imazalil has been widely used in citrus fruits such as lemons, oranges, and grapefruits. A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the detection of residual imazalil in citrus fruits. A monoclonal antibody (MoAb) generated to the synthetic imazalil hapten (EIT-0073)-protein conjugate was used. This assay was applied to lemon, orange, and grapefruit matrices for an imazalil analysis. The acceptable residue level for lemons, oranges, and grapefruits in Japan is 5 ppm. The matrix interference was minimized by direct dilution of the sample homogenate. No further cleanup was needed. The detection limit for imazalil in these citrus fruits was 0.1 ng/mL. The recovery of each fortified citrus fruit sample was >81.0%. The imazalil recovery measured by the proposed ELISA was compared to the recovery determined by a conventional HPLC. A good correlation was observed between the proposed ELISA and the HPLC. This proposed ELISA would be useful for monitoring for residual imazalil.     

Nuclear transfer in cattle : effect of linoleic acid-albumin on freezing sensitivity of enucleated oocytes

The effect of linoleic acid-albumin (LAA) supplementation to the media for IVM, enucleation, and activation on the developmental potential of bovine embryos produced by nuclear transfer (NT) into frozen-thawed cytoplasts was investigated. Blastomeres derived from morulae was placed in the perivitelline space of frozen-thawed cytoplasts, which were then fused by a DC pulse. The proportion of fused embryos was similar between groups with and without LAA (87 vs. 90%). The proportion of development to blastocysts of NT embryos derived from the media with LAA (14%) was higher than that without LAA (4%), indicating that LAA treatment of bovine oocytes during IVM, enucleation and activation can improve the ability of such cytoplasts after freezing and thawing to develop into blastocysts after NT.     

Molecular epidemiology of avian influenza viruses circulating among healthy poultry flocks in farms in northern Vietnam

Repeated epizootics of highly pathogenic avian influenza (HPAI) virus subtype H5N1 were reported from 2003 to 2005 among poultry in Vietnam. More than 200 million birds were killed to control the spread of the disease. Human cases of H5N1 infection have been sporadically reported in an area where repeated H5N1 outbreaks among birds had occurred. Subtype H5N1 strains are established as endemic among poultry in Vietnam, however, insights into how avian influenza viruses including the H5N1 subtype are maintained in endemic areas is not clear. In order to determine the prevalence of different avian influenza viruses (AIVs), including H5N1 circulating among poultry in northern Vietnam, surveillance was conducted during the years 2006-2009. A subtype H5N1 strain was isolated from an apparently healthy duck reared on a farm in northern Vietnam in 2008 and was identified as an HPAI. Although only one H5N1 virus was isolated, it supports the view that healthy domestic ducks play a pivotal role in maintaining and transmitting H5N1 viruses which cause disease outbreaks in northern Vietnam. In addition, a total of 26 AIVs with low pathogenicity were isolated from poultry and phylogenetic analysis of all the eight gene segments revealed their diverse genetical backgrounds, implying that reassortments have occurred frequently among strains in northern Vietnam. It is, therefore, important to monitor the prevalence of influenza viruses among healthy poultry between epidemics in an area where AIVs are endemic.     

Suspected panosteitis in a crossbred calf

A male crossbred calf developed a limp and pain upon deep pressure on the right hind limb and the right forelimb. The radiographic findings of affected limbs and pathological findings of bone biopsy were similar to those observed in canine panosteitis. This is the first case of suspected panosteitis reported in cattle.     

Pulmonary adenosquamous carcinoma in a dog

A mass that developed in the lung of a 10-year-old mixed-breed dog was pathologically examined. Histopathological examination showed papillary and tubular growth of glandular epithelium-like cells in some areas and growth of squamous cells arranged in nests in other areas, showing coexistence of adenocarcinoma and squamous cell carcinoma in a lung tumour. Immunohistochemical staining with anti-keratin-cytokeratin antibody was strongly positive for cytoplasms in both components. Electron microscopically, the neoplastic cells of the adenocarcinoma component had features of glandular cells, with microvilli, numerous free ribosomes, large round secretory granules and intercellular desmosomes. Non-keratinized squamous cells had tonofilaments and intercellular desmosomes. These findings led to the diagnosis of primary adenosquamous carcinoma, which demonstrates phenotypic profiles characteristic of both epidermal keratinocytes and glandular epithelium.     

Effects of orvus ES paste on canine spermatozoal longevity after freezing and thawing

Because of weak resistance of canine sperm to freezing, an applicable method of preparing canine frozen semen has not yet been established. We added various concentrations of Orvus ES Paste (OEP) to egg yolk Tris-fructose citrate, and investigated its effectiveness on survival of spermatozoa. Addition of 0.5-1.0% OEP to the extender for freezing canine semen was effective in prolonging post-thaw survival of spermatozoa.     

Genetic analysis of phosphoprotein and matrix protein of rabies viruses isolated in Brazil

To investigate the genetic characteristics of phosphoprotein (P) and matrix protein (M) genes of variable rabies virus (RV) prevalent in Brazil, the authors genetically characterized the P and M genes from 30 Brazilian RV field isolates. Phylogenetic analysis based on the P and M genes revealed the presence of six RV variants that consisted primarily of three insectivorous bats, the vampire bat, dog and fox in Brazil. Specific amino acid substitutions corresponding to these phylogenetic lineages were observed, with Asp(42) and Glu(62) in the P protein found to be characteristic of Brazilian chiroptera- and carnivora-related RVs, respectively. Amino acid sequence motifs predicted to associate with a viral function in the P and M proteins were conserved among Brazilian RV variants.     

Magnesium oxide nanoparticles induce systemic resistance in tomato against bacterial wilt disease

Bacterial wilt is a serious problem affecting many important food crops. Recent studies have indicated that treatment with biotic or abiotic stress factors may increase the resistance of plants to bacterial infection. This study investigated the effects of magnesium oxide nanoparticles (MgO NP) on disease resistance in tomato plants against Ralstonia solanacearum, as well as its antibacterial activity. The roots of tomato seedlings were inoculated with R. solanacearum and then immediately treated with MgO NP; the treated plants showed very little inhibition of bacterial wilt. In contrast, when roots were drenched with a MgO NP suspension prior to inoculation with the pathogen, the incidence of disease was significantly reduced. Rapid generation of reactive oxygen species such as O₂^?˙ radicals was observed in tomato roots treated with MgO NP. Further O₂^?˙ was rapidly generated when tomato plant extracts or polyphenols were added to the MgO NP suspension, suggesting that the generation of O₂^?˙ in tomato roots might be due to a reaction between MgO NP and polyphenols present in the roots. Salicylic acid‐inducible PR1, jasmonic acid‐inducible LoxA, ethylene‐inducible Osm, and systemic resistance‐related GluA were up‐regulated in both the roots and hypocotyls of tomato plants after treatment of the plant roots with MgO NP. Histochemical analyses showed that β‐1,3‐glucanase and tyloses accumulated in the xylem and apoplast of pith tissues of the hypocotyls after MgO NP treatment. These results indicate that MgO NP induces systemic resistance in tomato plants against R. solanacearum.     

Influence of Amylose Content on Cookie and Sponge Cake Quality and Solvent Retention Capacities in Wheat Flour

Reduced amylose wheat (Triticum æstivum L.) produces better quality noodles and bread less prone to going stale, while little is known about the relationships between amylose content and the quality of soft wheat baking products such as sugar snap cookies (SSC) and Japanese sponge cakes (JSC). Near‐isogenic lines developed from wheat cultivar Norin 61, differing in their level of granule‐bound starch synthase (Wx protein) activity, were used to produce wheat grains and ultimately flours of different amylose contents. These were tested with regard to their effect on soft wheat baking quality and solvent retention capacities (SRC). Amylose content was strongly correlated to cookie diameter (r = 0.969, P < 0.001) and cake volume (r = 0.976, P < 0.001), indicating that the soft wheat baking quality associated with SSC diameter and JSC volume were improved by an incremental increases in amylose content. Among the four kinds of SRC tests (water, sodium carbonate, sucrose and lactic acid), the water SRC test showed the highest correlation with amylose content, SSC diameter, and JSC volume. When the regression analysis was conducted between the nonwaxy and partial waxy isogenic lines that are available in commercial markets, only water SRC was significantly correlated to amylose content (r = –0.982, P < 0.001) among of four SRC tests. This suggests that, unlike udon noodle quality, high‐amylose content is indispensable in improving soft wheat baking quality, a process requiring less water retention capacity.     

Effect of Growing Environment of Soft Wheats on Amylose Content and Its Relationship with Cookie and Sponge Cake Quality and Solvent Retention Capacity

The effect of growing environments of soft wheat on amylose content and its relationship with baking quality and solvent retention capacities (SRC) was investigated. Near‐isogenic soft wheat lines of Norin 61 differing in granule‐bound starch synthase (Wx protein) activity and grown in three different regions of Japan: Hokkaido (spring‐sown) for 2006 and 2007, Kanto (autumn‐sown), and Kyushu (autumn‐sown) for 2007 were evaluated. Spring‐sown samples produced grains of greater protein content (10.9–12.4%) than autumn‐sown samples (7.3–9.1%). In contrast, spring‐sown samples of 2007 with higher maturing temperature had lower amylose content (25.5% for Norin 61) compare to the autumn‐sown and spring‐sown samples of 2006 (27.6–28.4% for Norin 61). Amylose content was strongly correlated to sugar snap cookie (SSCD) diameter (r = 0.957–0.961; n = 10, all samples; P ≤ 0.001, r = 0.701–0.976; n = 7 partial waxy and nonwaxy samples; and Japanese sponge cake (JSCV) volume r = 0.971–0.993; n = 10; P≤ 0.001, r = 0.764–0.922; n = 7 partial waxy and nonwaxy samples), regardless of seeding season and growing conditions. The strength of the JSVC‐amylose relationship (slope) was similar among the three regions, whereas the strength of the SSCD‐amylose relationship was slightly weaker for spring‐sown samples and slightly stronger for partial waxy and nonwaxy autumn‐sown samples. Among of the four solvents (water, solutions of sodium carbonate, sucrose, or lactic acid), water‐SRC showed the greatest correlation to amylose content (r = –0.969 to –0.996; n = 10; P ≤ 0.001, r = –0.629 to –0.983; n = 7 partial waxy and nonwaxy samples), indicated that amylose content can be accurately estimated from the water‐SRC within the samples from the same grown environment.