Use of an enzyme-linked immunosorbent assay in a bovine brucellosis eradication program

An enzyme-linked immunosorbent assay (ELISA) was developed and was compared with the complement fixation test (CFT) in a bovine brucellosis eradication program. The ELISA detected significantly more reactors than the CFT in both strain 19 vaccinated infected herds (1.79% versus 1.14%) and non-vaccinated infected herds (4.2% versus 3.59%) but not in either vaccinated or non-vaccinated brucella-free herds. The specificity for both tests in brucella-free herds was greater than 0.998. The specificity and sensitivity of the ELISA were compared with those of 3 other tests (the Rose Bengal test; the indirect haemolysis test [IHLT] and the CFT) on serum from 151 animals cultured at slaughter. The calculated specificity of the ELISA in this infected group was lower than for both the CFT and the IHLT (0.58 versus 0.67 versus 0.75). The sensitivity however was much greater (1.0 versus 0.73 versus 0.71). The value of the ELISA when used in an eradication program is discussed.     

Evaluation of an enzyme linked-immunosorbent assay for the diagnosis of Brucella ovis infection in rams

A simple enzyme linked immunosorbent assay (ELISA) was developed for the serological diagnosis of Brucella ovis infections in rams. Serums from brucellosis accredited-free flocks and flocks known to be infected with B. ovis were tested and the results correlated with warm complement fixation (CF) test and bacteriological examination of semen. Both the ELISA and the CF test detected 0.5% false positive reactions in rams from clinically negative flocks. However the ELISA detected significantly more positive reactors in infected flocks and the CF test failed to detect some rams excreting B. ovis. The ELISA proved to be a valuable test in eradicating brucellosis from infected flocks.     

The development of an assay for resistance to Phytophthora cambivora in almond rootstocks using shoots excised from tissue cultures

The pathogenicity of Phytophthora cambivora to Mission almond seedlings was confirmed by twig inoculation in situ. An excised twig assay using four isolates of P. cambivora showed almond cvs Mission and Chellaston to be susceptible, whereas peach cv. Nemaguard, which is commonly used as a rootstock for almond, was resistant. P. cambivora isolates 179 and 108, the most and least aggressive, respectively, were selected for further studies. The excised twig assay was modified for screening micropropagated shoots in vitro for response to P. cambivora. Defoliated shoots were placed upright in agar medium colonized with the fungus, and development of necrosis was assessed over a period of 2-3 days. Shoots of Chellaston developed extensive necrosis and those of Nemaguard developed little necrosis, whereas shoots of hybrid selections from peach x almond crosses were generally intermediate in response. This method permits rapid and efficient screening of large amounts of material, and would be a useful adjunct to conventional almond breeding programmes.     

Use of phages for identifying the citrus canker bacterium Xanthomonas campestris pv. citri in Taiwan

Phages CP115 and CP122, which were isolated from canker lesions on grapefruit and Liucheng sweet orange, respectively, showed a high degree of specificity with respect to lysis of test bacterial strains. When used jointly, they lysed 135 (97·8%) out of 138 Xanthomonas campestris pv. citri strains isolated from the canker lesions on leaves, twigs, and fruits of various citrus species, cultivars, and hybrids grown throughout Taiwan, but they did not lyse other X. campestris pathovars and other phytopathogenic bacteria, nor other bacteria isolated from soil, clinical or environmental samples. Of 252 CP115/CP122-sensitive and 78 CP115/CP122-resistant bacterial strains with colony characteristics typical of or similar to those of X. campestris pv. citri , isolated from canker lesions of various citrus plants in diverse growing regions in Taiwan, 250 (99·2%) and 76 (97·4%) strains were pathogenic and non-pathogenic, respectively, when inoculated into Liucheng sweet orange or Mexican lime. Thus, phages CP115 and CP122, when used jointly, appear to be applicable for identifying X. campestris pv. citri in Taiwan.     

Inhibition of growth hormone secretagogue receptor antagonist, [D-Lys-3]-GHRP-6, in adipogenesis of ovine and rat adipocytes

Ghrelin and growth hormone secretagogues receptor (GHS‐R or ghrelin receptor) have been reported as being one of the factors of adipogenesis in adipocytes. To investigate the involvement of ghrelin and GHS‐R in adipocytes, the effect of the GHS‐R antagonist, [D‐Lys‐3]‐GHRP‐6 (His‐D‐Trp‐D‐Lys‐Trp‐D‐Phe‐Lys‐NH₂), on the process of adipogenesis in ovine and rat adipocytes was evaluated. [D‐Lys‐3]‐GHRP‐6 (10^?7 mol/L) significantly inhibited adipogenic differentiation of ovine and rat preadipocytes prepared from adipose tissues. The level of peroxisome proliferator activated receptor (PPAR)‐γ2 mRNA, an adipogenic marker, was decreased during the differentiation of adipocytes treated with [D‐Lys‐3]‐GHRP‐6 for 10 days. Ghrelin stimulated adipogenesis, also causing an increment of glycerol‐3‐phosphate dehydrogenase and upregulation of PPAR‐γ2. Furthermore, the antilipolytic effect of ghrelin was attenuated by treatment with [D‐Lys‐3]‐GHRP‐6 in both types of isolated adipocytes. Overall, the results of the present study highlight that GHS‐R in adipogenesis can be blocked by treatment with [D‐Lys‐3]‐GHRP‐6.     

Allelopathic potential of K21, selected as a promising allelopathic rice

The agronomic characteristics of a newly bred potent allelopathic rice, K21, including morphological appearance, resembled its female parent, Dongjinbyeo, except for 8 days' earlier heading and 9 cm shorter plant height. The water extract of K21 showed a greater inhibitory effect on the shoot and root growth of barnyardgrass than its female parent, but less than the male parent, Kouketsumochi. At the highest extract concentration of 5.0%, the inhibitory effect of K21 on the shoot growth of barnyardgrass was ≈ 64%, similar to its male parent. The dry weight of the barnyardgrass shoot was reduced by 13.0% when grown with K21 and 30.2% when grown with its male parent. There was no inhibitory effect on the shoot dry weight of barnyardgrass when grown with the female parent. The activity of cinnamic acid 4-hydroxylase in K21 was ≈ 1.8-fold higher than that of the female parent, but significantly lower than that of the male parent. The ρ-coumaric acid content in K21 was ≈ 60% higher than that of the female parent, but it was about half of the male parent. The total amount of phenolic compounds in K21 was ≈ 11.5% higher than in the female parent, but about half of the male parent. The results obtained suggest that the newly bred K21 contains desirable allelopathic traits derived from the male parent, Kouketsumochi, and acceptable agronomic traits from its female parent, Dongjinbyeo.     

Comparative in vitro effects of closantel and selected β-ketoamide anthelmintics on a gastrointestinal nematode and vertebrate liver cells

PNU-87407 and PNU-88509, β-ketoamide anthelmintics that are structurally related to each other and to the salicylanilide anthelmintic closantel, exhibit different anthelmintic spectra and apparent toxicity in mammals. The basis for this differential pharmacology was examined in experiments that measured motility and adenosine triphosphate (ATP) levels in larval and adult stages of the gastrointestinal nematode, Haemonchus contortus , and in a vertebrate liver cell line and mitochondria. PNU-87407 and PNU-88509 both exhibited functional cross-resistance with closantel in larval migration assays using closantel-resistant and -sensitive isolates of H. contortus . Each compound reduced motility and ATP levels in cultured adult H. contortus in a concentration- and time-dependent manner; however, motility was reduced more rapidly by PNU-88509, and ATP levels were reduced by lower concentrations of closantel than the β-ketoamides. Tension recordings from segments of adult H. contortus showed that PNU-88509 induces spastic paralysis, while PNU-87407 and closantel induce flaccid paralysis of the somatic musculature. Marked differences in the actions of these compounds were also observed in the mammalian preparations. In Chang liver cells, ATP levels were reduced after 3 h exposures to 0.25 μ M PNU-87407, 1 μ M closantel or 10 μ M PNU-88509. Reductions in ATP caused by PNU-88509 were completely reversible, while the effects of closantel and PNU-87407 were irreversible. PNU-87407, closantel and PNU-88509 uncoupled oxidative phosphorylation in isolated rat liver mitochondria, inhibiting the respiratory control index (with glutamate or succinate as substrate) by 50% at concentrations of 0.14, 0.9 and 7.6 μ M , respectively.