Development and evaluation of a loop-mediated isothermal amplification assay for detection of Erwinia amylovora based on chromosomal DNA

A reliable and rapid pathogen detection protocol that utilizes loop-mediated isothermal amplification (LAMP) was developed for detection of Erwinia amylovora, the casual agent of fire blight. The six LAMP primers applied were derived from the highly conserved fragment of the chromosomally amsH gene. Despite the proposed LAMP as well as nested PCR presenting equal values of sensitivity (2?×?10¹?CFU/ml or more) for pure cultures, as compared with conventional PCR (2?×?10³?CFU/ml), both methods were together superior. The specificity assay also showed that the LAMP protocol is species-specific for detection of E. amylovora even in inter-species analysis. Meanwhile, when all 208 naturally infected samples were examined, the specificity value of LAMP was 84%, while conventional and nested PCR could detect only 59% and 73% of the whole collection. Significantly, an independent behaviour versus host plant as well as each strain origin was also observed regarding the current LAMP method as well as other two PCR-based methods. All the results, overall, indicated that the LAMP offers an interesting novel and convenient assay format for the quick and specific chromosomal detection and diagnostic tool of recognition of E. amylovora and therefore presents an alternative to PCR-based assays.     

Combined or individual effects of dietary magnesium and selenium nanoparticles on growth performance,immunity, blood biochemistry and antioxidant status of Asian seabass (Lates calcarifer) reared in freshwater

This study was conducted to investigate effects of and interactions between nanoselenium (NanoSe) and nanomagnesium (NanoMg) on growth, humoral immunity, serum biochemistry and antioxidant capacity of juvenile Asian seabass Lates calcarifer reared in freshwater. Four groups of fish with an average weight of 32.78 ± 2.23 g were fed one of the experimental diets for 6 weeks: (a) control (basal diet); (b) NanoSe (basal diet + 4 mg NanoSe/kg diet); (c) NanoMg (basal diet + 500 mg NanoMg/kg diet); and (d) combination (basal diet + 4 mg NanoSe/kg diet + 500 mg NanoMg/kg diet). Fish fed with NanoSe‐supplemented diets (NanoSe and combination) showed higher weight gain, specific growth rate and feed intake. The combination of NanoSe and NanoMg enhanced the immune response. Also, fish fed on combination diet showed higher serum alanine aminotransferase and aspartate transaminase levels but lower glucose concentration. The activities of liver superoxide dismutase, glutathione peroxidase and catalase were not different among all groups, but liver malondialdehyde level was lower in fish fed diets supplemented with NanoSe and/or NanoMg. It could be concluded that NanoSe improved growth performance; the combination of both NanoSe and NanoMg enhanced humoral immunity; and NanoSe and/or NanoMg improved antioxidant capacity in Asian seabass.     

Effects of dietary crude protein and electrolyte balance on production parameters and blood biochemicals of laying hens under tropical summer condition

The objective of this study was to determine the effects of different levels of crude protein (CP) and dietary electrolyte balance (DEB) on productive performance, egg quality, and blood biochemical parameters of laying hens reared during hot summer months. The average minimum and maximum room temperatures recorded from 32 to 42 weeks of age were 27.2 and 36.5 °C, respectively, with relative humidity ranging from 48 to 56 %. A total of 144 Lohmann LSL-Lite laying hens were assigned to each of four dietary treatments with six replicate cages of six birds. The treatments were as follows: CP 15.4 % + DEB 165 mEq/kg (group 1), CP 15.4 % + DEB 250 mEq/kg (group 2), CP 13.9 % + DEB 165 mEq/kg (group 3), and CP 13.9 % + DEB 250 mEq/kg (group 4). In groups 3 and 4, feed conversion ratio and blood uric acid were decreased (P?<?0.05); but the plasma triglyceride level was increased (P?<?0.05) compared to groups 1 and 2. Low-CP diet with DEB 250 (group 4) also induced significant increases of average egg production and eggshell thickness compared to group 1. Blood pH, Na⁺, and HCO₃ ^? values were also higher (P?<?0.05) in groups 2 and 4 than in the other groups. These results indicate that, under tropical conditions, using a DEB of 250 mEq/kg achieves a correction of the lay-induced metabolic acidosis and induces a positive effect on eggshell quality when the hens were fed low-protein diet.     

Responses of ‘Petopride’ processing tomato to partial rootzone drying at different phenological stages

Partial rootzone drying (PRD) is a water-saving irrigation practice which involves watering only part of the rhizosphere at each irrigation with the complement left to dry to a pre-determined level. The effect of PRD, applied at different phenological stages, on yield, fruit growth, and quality of the processing tomato cv. ‘Petopride’ was studied in this experiment. The treatments were: daily full irrigation (FI) on both sides of the root system considered as the control, and PRD treatments applied at three phenological stages. These were: during the vegetative stage until the first truss was observed (PRD_VS–FT), from the first truss to fruit set (PRD_FT–FS), and from fruit set to harvest (PRD_FS–H). In some occasions, leaf xylem water potential was lower in each PRD period than in FI. Number of fruits, total fresh and dry weight of fruit per plant, harvest index, and fruit growth were lower in PRD_FT–FS and PRD_FS–H plants than in FI and PRD_VS–FT plants. However, irrigation water use efficiency, on a dry weight basis, was the same among the treatments. For PRD_FT–FS and PRD_FS–H treatments, mean fresh weight of fruit and fruit water content were reduced and dry matter concentration of cortex and total soluble solids concentration of fruit increased compared with FI and PRD_VS–FT treatments. Incidence of blossom-end rot was the same among PRD_VS–FT, PRD_FS–FH, and FI fruit, but it was higher in PRD_FT–FS fruit. Fruit skin colour was the same among treatments. Total dry weight of fruit per plant decreased by 23% for PRD_FT–FS and by 20% for PRD_FS–H relative to FI. Fruit quality improvement in PRD_FS–H could compensate for the reduction in total dry weight of fruit where water is expensive for tomato production. But an economical analysis would be needed to substantiate this. PRD from the first truss to fruit set is not recommended because of the high incidence of blossom-end rot. An erratum to this article can be found at     

The effect of safranal on histamine (H1) receptors of guinea pig tracheal chains

The effect of three concentrations of safranal on histamine (H₁) receptors was tested on two groups of tracheal chains incubated with: 1) indomethacin, and 2) indomethacin, propranolol and atropine (n = 6). The EC₅₀ (effective concentration of histamine causing 50% of maximum response) obtained in the presence of chlorpheniramine and all concentrations of safranal in both groups were significantly greater than those of saline (p < 0.05 to p < 0.001). The EC₅₀ obtained in the presence of all concentrations of safranal and maximum response of its two higher concentrations (1.25 and 2.5 μg/mL) in group 2 were greater than in group 1 (p < 0.05 to p < 0.001).     

Assessment of the biological control capability of <Emphasis Type="Italic">Hippodamia variegata</Emphasis> (Col.: Coccinellidae) using functional response experiments

Lady beetles are among the most successful predators of aphids in different environments. The functional responses of different life stages of Hippodamia variegata (Goeze) towards cotton aphids were examined in two different set-ups, a two-dimensional Petri dish set-up with detached leaves and a three-dimensional set-up with whole plants. In addition, the functional responses in two-dimensional set-ups towards cotton aphids and the pea aphids were compared. H. variegata exhibited a functional type II response to both cotton aphids and pea aphids irrespective of life stage and spatial scale of the set-up. Females and fourth instars generally had higher search rates than third instars whereas handling times were consistently lower for the fourth instar stage compared with the preceding juvenile stage and with females. The spatial dimensions did not have any significant influence on the functional responses towards cotton aphids, except for third instars which in the three-dimensional set-up reduced their search rate and increased their handling time. Fourth instars reacted in the same way to both Aphis gossypii and Acyrthosiphon pisum whereas the functional response parameters for third instars and females were significantly different on the two prey species. Our study, a part of this first-step evaluation of H. variegata as a biocontrol agent against A. gossypii and A. pisum under field condition, suggest that the voracity of all tested stages of H. variegata towards both aphid species hold good promises for a use of especially fourth instars and females in inundative biocontrol.     

Gene expression of endothelin-1 and its receptors in the heart of broiler chickens with T(3)-induced pulmonary hypertension

To investigate the effect of T₃-induced pulmonary hypertension on endothelin (ET) production and genes expression of ET-1, ET_A and ET_B receptors (ET_AR and ET_BR) during rearing, semiquantitative RT-PCR and enzyme immunometric assay were performed in the heart ventricles and serum, respectively. The ET-1 and its receptor genes were expressed in the right and left ventricles of control and T₃-treated broilers at 12, 28 and 49 days of age. There were significant (P < 0.05) reductions of the relative amounts of ET-1 (in both ventricles) and ET_AR (in the right ventricle) mRNAs at 28 and 49 days of age, in T₃-treated broilers compared to controls. The relative amounts of ET_BR mRNA in the right and left ventricles did not significantly differ between control and T₃-treated broilers at any age. The serum level of ET was significantly (P < 0.05) increased in T₃-treated chickens at 28 and 49 days of age when compared with that of the control. It is concluded that ET-1, ET_AR and ET_BR genes are normally expressed in the heart ventricles of broilers. It is likely that increased serum level of ET and decreased ET-1/ET_AR genes expression in the ventricles are involved in the heart dysfunction of broiler chickens with developmental pulmonary hypertension.