Development of an enzyme-linked immunosorbent assay for the pyrethroid cypermethrin

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of cypermethrin was developed. Two haptens, the trans- and cis-isomers of 3-[(+/-)-cyano-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarbonyloxy]methyl]phenoxyacetic acid, were conjugated with thyroglobulin as immunogens. Four antisera were generated and screened against six different coating antigens. The assay that was the most sensitive for cypermethrin was optimized and characterized. The IC(50) for cypermethrin was 13.5 +/- 4.3 microg/L, and the lower detection limit (LDL) was 1.3 +/- 0.5 microg/L. This ELISA had relatively low cross-reactivities with other major pyrethroids, such as deltamethrin, phenothrin, resmethrin, fluvalinate, and permethrin. Methanol was found to be the best organic cosolvent for this ELISA, with an optimal sensitivity observed at a concentration of 40% (v/v). The assay parameters were unchanged at pH values between 5.0 and 8.0, whereas higher ionic strengths strongly suppressed the absorbances. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction was applied to various domestic and environmental water samples. The water samples, fortified with cypermethrin, were analyzed according to this method. Good recoveries and correlation with spike levels were observed.     

Salmonella enterica subspecies enterica serotype Typhimurium and Escherichia coli O86 in wild birds at two garden sites in south-west Scotland

Salmonella enterica subspecies enterica serotype Typhimurium and Escherchia coli O86:K61:NM are two bacteria that can cause outbreaks of mortality in garden birds visiting bird tables and other feeding stations. Two sites in south-west Scotland were monitored for the two organisms for 12 months. At site A, large numbers of birds fed throughout the year, and at site B smaller numbers of birds fed only in the winter months. Samples of composite faeces were collected from the feeding stations and screened for the organisms, and any dead birds were also screened. S Typhimurium definitive type (DT) 56 (variant) was found to be endemic at site A, and was recovered from 48 per cent of samples of composite faeces collected from the bird table, from 42 per cent of composite faeces from underneath a hanging feeder, and from 33 per cent of composite faeces from below a roost used by house sparrows; the organism was also isolated from the carcases of six wild birds found dead at the site. In contrast, S Typhimurium (DT41) was recovered only once at site B, from 2 per cent of the composite faeces from below a hanging feeder, and no dead birds were recovered from the site. E coli O86 was not recovered from the faeces collected from either site, but was isolated from a bird that died from trauma at site A.     

Host immune responses against hog cholera virus in pigs treated with an ionized alkali mineral complex

To determine the immune responses in pigs to hog cholera virus after treatment with an ionized alkali mineral complex (IAMC), 40 healthy pigs (28-32 days old) from a commercial swine farm were purchased and housed into 4 groups (n=10 each). All pigs were vaccinated intramuscularly (1 ml) with an attenuated live hog cholera virus (HCV, LOM strain) at 28-32 days old and challenged with a virulent hog cholera virus at 8 weeks after vaccination. Each group was treated with PowerFeel sprayed diet as 0.05% (w/w) in a final concentration (T-1, n=10), a diet mixed with SuperFeed as 3% (w/w) in a final concentration (T-2, n=10), or a diluted PowerFeel solution (1:500, v/v) as drinking water (T-3, n=10), respectively. A group (n=10) served as a non-treated control. Proportions of expressing CD2+ and CD8+ cells increased significantly (p<, 0.05) at 8-week post-application. Mean antibody titers of each group against HCV gradually increased to higher levels after vaccination and with challenge of the virulent virus. In conclusion, the IAMC-treated diets can be helpful for the improvement of growth in pigs with proper vaccination program, while the IAMC-treated diets have no effects on the clinical protection against hog cholera.     

Isolation of Neospora caninum from the brain of a naturally infected adult dairy cow

Neospora caninum was isolated from the brain of a 2-year-old dairy cow that had aborted confirmed N. caninum-infected fetuses on two occasions. The cow had an indirect fluorescent antibody titer of 1:1600 to N. caninum. The cow was killed 24 days after its second abortion and the brain was bioassayed for N. caninum in nude mice. Multifocal areas of perivascular cuffing and glial nodules were observed in the cerebrum and mesencephalon of the cow, but N. caninum was not identified in histological sections of the brain. All three nude mice inoculated with brain homogenate of the cow, developed emaciation and paralysis. Microscopical examination of the nude mice revealed systemic N. caninum infection with demonstrable tachyzoites in various organs. The parasites isolated from fresh mouse brain were transferred successfully into Vero cell cultures. PCR procedure on the purified tachyzoites obtained from the Vero cell cultures amplified the specific DNA sequence for N. caninum.     

Effect of soil water stress on the development of stone cells in pear (Pyrus pyrifolia cv. ‘Niitaka’) flesh

This study was carried out to investigate the cause of stone cell formation in pear (Pyrus pyrifolia cv. ‘Niitaka’) flesh. Potted plants grown in a glass house were subjected to water stress conditions without irrigation for 30 days from 30 days before full bloom (BFB treatment), full bloom (FB treatment) and 30 days after full bloom (AFB treatment). Control plants were drip-irrigated daily maintaining a soil matrix potential around −40 ± 5 kPa. The formation of stone cells in pear flesh increased in the FB treatment and AFB treatment plants and this tendency was sustained until the harvest season. Root activity was investigated 60 days after full bloom (DAFB) and the triphenyltetrazolium chloride (TTC) reduction potential, the formazan content and leaf water potential were investigated 30, 45, and 60 DAFB. Root activity decreased progressively due to the effect of water stress. Also, the Ca content in leaf and flesh was lower. The peroxidase activity was high in the flesh at the early stages of fruit growth and decreased at the late stages of fruit growth, and then a higher increase of peroxidase activity was observed in water-stressed fruit. The reduction in calcium content of leaf and fruit in plants under water stress may be related to the reduction of root activity and leaf water potential. The increase in peroxidase activity under water stress may be due to limited calcium absorption. Higher peroxidase activity may induce the accumulation of lignin in the cell wall and promote the formation of stone cells in pear flesh. We conclude that water stress condition during the early stages of fruit growth is one of several factors that determine the formation of stone cells in pear flesh.