Determination of spinosad and its metabolites in food and environmental matrices. 2. Liquid chomatography-mass spectrometry

A selective and sensitive method utilizing liquid chromatography-mass spectrometry (LC-MS) has been developed for determining residues of the natural insect control agent spinosad in several crop matrices that are difficult to analyze by HPLC with UV detection. The method determines the active ingredients (spinosyns A and D) and three minor metabolites (spinosyns B and K and N-demethylspinosyn D) in alfalfa hay, wheat hay, wheat straw, sorghum fodder, and corn stover. The analytes are extracted from the samples with an acetonitrile/water solution, and the extracts are purified by solid phase extraction with a C(18) disk and a silica cartridge. All five analytes are determined simultaneously in a single injection using positive atmospheric pressure chemical ionization LC-MS with selected ion monitoring. The average recoveries ranged from 69 to 96% with standard deviations ranging from 4 to 15%. The method has a validated limit of quantitation of 0. 01 microgram/g and a limit of detection of 0.003 microgram/g. The LC-MS method can also provide residue confirmation in addition to quantitation.     

The composite genome of the legume symbiont Sinorhizobium meliloti

The scarcity of usable nitrogen frequently limits plant growth. A tight metabolic association with rhizobial bacteria allows legumes to obtain nitrogen compounds by bacterial reduction of dinitrogen (N2) to ammonium (NH4+). We present here the annotated DNA sequence of the alpha-proteobacterium Sinorhizobium meliloti, the symbiont of alfalfa. The tripartite 6.7-megabase (Mb) genome comprises a 3.65-Mb chromosome, and 1.35-Mb pSymA and 1.68-Mb pSymB megaplasmids. Genome sequence analysis indicates that all three elements contribute, in varying degrees, to symbiosis and reveals how this genome may have emerged during evolution. The genome sequence will be useful in understanding the dynamics of interkingdom associations and of life in soil environments.     

Quantitative trait loci influencing fruit-related characteristics of tomato grown in high-temperature conditions

The objective of this study was to identify molecular markers linked to fruit-related traits in the tomato subjected to high temperatures. In total, 160 F₂ plants derived from a cross between a heat-tolerant breeding line, CL5915-93D4-1-0-3 (Solanum esculentum), and a heat-sensitive wild accession, L4422 (S. pimpinellifolium), were grown in a greenhouse. Six traits including fruit number, fruit weight, brix, seed number, fruit setting, and flower number were scored. The distributions of fruit number, fruit set, flower number, and seed number were skewed towards heat susceptibility which is known to be characteristic of L4422. Polymorphic bands were generated by PCR-derived methods of RAPD, ISSR and AFLP Polymorphism, the segregation ratio, and distribution over the genome of the above 3 markers were compared. Ten linkage groups, ranging 20.6–151.6 cM in size, were constructed with 62 informative markers spanning a total of 776.3 cM. Fruit-related quantitative trait loci (QTLs) were non-randomly distributed in the tomato genome. For the 6 traits investigated, 21 QTLs were dispersed on linkage groups 2–5. The genetic effects of the various QTLs were differently exhibited, in our study we have respectively found from 10.5% to 30.2% of the variation explained by the QTL for flower number (FRN4) and brix (BX2). Thirteen QTL-mapped markers were unique to 1 trait, and 4 markers were linked to more than 1 trait. Among them, QTLs linked to the I868-470 marker had effects on fruit weight and brix, and a significant positive correlation between these 2 traits was noted (r = 0.35, P < 0.05). Thus, the I868-470 marker may have the potential for simultaneous selection of high fruit weight and brix. These markers also allowed us to align genome linkage maps across distantly related species and to reveal the co-localization between these QTLs and major genes.     

Clinical comparison of recovery from total intravenous anesthesia with propofol and inhalation anesthesia with isoflurane in dogs

The characteristics of recovery from total intravenous anesthesia (TIVA) with propofol and inhalation anesthesia with isoflurane was clinically compared in 149 client-owned dogs that anesthetized for surgical or diagnostic procedures. In all dogs, anesthesia was induced with an intravenous injection of propofol following premedication with acepromazine or diazepam. As a result, 58 dogs anesthetized with propofol-TIVA showed slower but smoother recovery than 91 dogs anesthetized with isoflurane anesthesia. The dogs stood at 34.5 +/- 19.3 and 27.7 +/- 17.2 min after propofol-TIVA and isoflurane anesthesia, respectively. Adverse effects, including hypersalivation, neurologic excitement (paddling, muscle tremor/twitching, opisthotonos) and vomiting/retching, were observed in similar infrequent incidences during the recovery from both anesthetic protocols. Propofol-TIVA is suggested to be an alternative anesthetic protocol for canine practice.     

Standardizing catch and effort data of the Taiwanese distant-water longline fishery in the western and central Pacific Ocean for bigeye tuna, Thunnus obesus

Catch per unit effort (CPUE) is often used as an index of relative abundance in fisheries stock assessments. However, the trends in nominal CPUE can be influenced by many factors in addition to stock abundance, including the choice of fishing location and target species, and environmental conditions. Consequently, catch and effort data are usually ‘standardized’ to remove the impact of such factors. Standardized CPUE for bigeye tuna, Thunnus obesus, caught by the Taiwanese distant-water longline fishery in the western and central Pacific Ocean (WCPO) for 1964–2004 were derived using three alternative approaches (GLM, GAM and the delta approach), and sensitivity was explored to whether catch-rates of yellowfin tuna and albacore tuna are included in the analyses. Year, latitude, and the catch-rate of yellowfin explained the most of the deviance (32–49%, depending on model configuration) and were identified consistently among methods, while trends in standardized catch-rate differed spatially. However, the trends in standardized catch-rates by area were found to be relatively insensitive to the approach used for standardization, including whether the catch-rates of yellowfin and albacore were included in the analyses.     

Two reported cytotypes of the emergent orchid model species <Emphasis Type="Italic">Erycina pusilla</Emphasis> are two different species

Each species is characterized by a specific set of chromosomes, which is described as the chromosome portrait or karyotype. In general, such a karyotype is the same for all individuals in the population. An exception to that rule has recently been found in the orchid Erycina pusilla, which has been reported to have two cytotypes with chromosome numbers of 2n = 10 and 2n = 12. Here, we examined the karyotypes of the two cytotypes and found differences in arm ratios and heterochromatin patterns as well as in the presence of satellite chromosomes and in the number and location of rDNA and telomeric repeat sites. These differences are extensive and would have required multiple chromosome rearrangements to generate the differences between the two karyotypes. We also found that F1 hybrids between the parents with the two different chromosome numbers resulted in sterile offspring, in accordance with our previous findings. The combination of hybrid sterility and extensively rearranged chromosomes supports the hypothesis that these two reported cytotypes are, in fact, two different species.     

Surgical outcome and ocular complications of evisceration and intraocular prosthesis implantation in dogs with end stage glaucoma: a review of 20 cases

Surgical outcome and complications following evisceration and implantation of intraocular silicone prosthesis (ISP) in 20 canine eyes with end-stage glaucoma were analyzed, including clinical signs, complications, cosmetic appearance, and owners' responses. The mean postoperative follow-up time was 312 days. Major short-term surgical complications were central corneal ulceration (6/20) and infections (3/20), while long-term complication was keratoconjunctivitis sicca (KCS) (2/20). All patient owners (100%) showed satisfied with the surgical outcome and postoperative cosmetic effect including 85% of the owners gave an excellent or good rating of satisfaction. No more medication needed for long term control except the KCS cases. With careful case selection and post-operative care, evisceration with ISP implantation proves a good and safe surgical procedure with minimal complications for end stage glaucoma.     

Comparison of ambisense m RNA of watermelon silver mottle virus with other tospoviruses

ABSTRACT Double-stranded genomic RNAs (dsRNAs) extracted from Chenopodium quinoa infected with watermelon silver mottle virus (WSMV) were similar to those of tomato spotted wilt virus (TSWV, serogroup I) and impatiens necrotic spot virus (INSV, serogroup III), except that the S dsRNA of WSMV is 0.75 and 0.6 kbp longer than those of TSWV and INSV, respectively. The complete nucleotide sequence of the genomic M RNA of WSMV was determined from cDNA clones generated from separated M dsRNA. The M RNA is 4,880 nucleotides in length with two open reading frames (ORFs) in an ambisense organization. The M RNA-encoded nonstructural (NSm) ORF located on the viral strand encodes a protein of 312 amino acids (35 kDa), and the G1/G2 ORF located on the viral complementary strand encodes a protein of 1,121 amino acids (127.6 kDa). The RNA probe corresponding to the NSm or G1/G2 ORF of WSMV failed to hybridize with the M dsRNAs of TSWV and INSV. Comparison of M and S RNAs of WSMV, TSWV, INSV, and peanut bud necrosis virus (PBNV, serogroup IV) revealed a consensus sequence of eight nucleotides of 5'-AGAGCAAU...-3' at their 5' ends and 5'-...AUUGCUCU-3' at their 3' ends. The low overall nucleotide identities (56.4 to 56.9%) of the M RNA and the low amino acid identities of the NSm and G1/G2 proteins (30.5 to 40.9%) with those of TSWV and INSV indicate that WSMV belongs to the Tospovirus genus but is phylogenetically distinct from viruses in serogroups I and III. The M RNA of WSMV shares a nucleotide identity of 79.6% with that of PBNV, and the two viruses share 83.4 and 88.7% amino acid identities for their NSm and G1/G2 proteins, respectively. It is concluded that they are two related but distinct species of serogroup IV. In addition to the viral or viral complementary full-length M RNA, two putative RNA messages for the NSm gene and the G1/G2 gene, 1.0 and 3.4 kb, respectively, were detected from the total RNA extracted from WSMV-infected tissue of Nicotiana benthamiana. The 1.0- and 3.4-kb RNAs were also detected in the viral RNAs extracted from purified nucleocapsids, suggesting that the putative messages of the M RNA of WSMV can also be encapsidated by the nucleocapsid protein.     

Callus induction and plant regeneration from lateral shoots of herbaceous bamboo Mniochloa abersend

Callus induction and plant regeneration of Mniochloa abersend via lateral shoots were conducted in this study. Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/L naphthaleneacetic acid (NAA) was effective for compact callus induction. Remarkably, calli on the MS medium with 0.1 mg/L 2,4-D yielded the highest folds of proliferation (8.01), and showed a high potential capacity to differentiate 1 year after subculture. In addition, the compact calli possessed 100% differentiation rate and generated more shoots that were green and strong in 1.0 mg/L kinetin and 1.0 mg/L NAA. Vigorous roots were generated in the 1/2MS supplemented with 0.5 mg/L indole-3-butyric acid, and the resultant plantlets exhibited 90% survival rate after they were hardened and transplanted. The established regeneration system of M. abersend provides a promising platform for bamboo gene function study.     

Effects of sugar cane extract on pseudorabies virus challenge of pigs

This experiment aimed to evaluate the efficacy of sugar cane extract (SCE) on the modulation of porcine immunity against pseudorabies virus (PrV) infection. Twelve-week-old experimental pigs were fed with SCE (500 mg/kg of body weight per day) for 3 days and challenged with PrV (2 x 10(5) TCID(50)) on the second day. Pigs that were only challenged with PrV and without SCE-treatment served as controls. The leukocyte functional assays were performed on the 7th and 14th day post-PrV challenge. Our results showed a significant enhancement (P<0.05) of natural killer cytotoxicity, lymphocyte proliferation, phagocytic function of monocytes, and interferon-gamma (IFN-gamma) production of CD4(+) and gammadelta T cells in the SCE-treated pigs compared with the controls. In addition, SCE administration reduced the severity of clinical signs and brain lesion in the course of disease in PrV-challenged pigs. SCE-treated pigs showed a 12% growth enhancement compared with untreated controls. SCE administration had an immunostimulating effect on porcine immunity that may subsequently enhance protective activities against PrV infection which may be extensively applied in field for the prevention of infections.