Temperature shifts induce adaptive changes in the physical state of carp (Cyprinus carpio L.) erythrocyte plasma membranes in vitro

Blood, freshly collected from warm- and cold-acclimated carp, Cyprinus carpio L., was cooled to 5°C for 4h or warmed to 25°C for 4h, respectively, and the fluorescence anisotropy of washed red blood cells was recorded using the fluorescent dye 3-(p-(6-phenyl-1,3,5-hexatrienyl) phenyl propionic acid [DPH-PA] (which is restricted to the outer leaflet of the plasma membrane) before and after the temperature shift. Despite individual variation, the plasma membrane of cold-exposed erythrocytes became more fluid while that of warm-exposed cells became more rigid following the temperature shift. This response was rapid and reversible. Cold-exposed cells from warm-acclimated fish became more fluid within 40–60 minutes and reverted to their original fluidity within the same time on warming, at a rate of 1°C/min; erythrocytes, from cold-adapted carp displayed an opposite change in fluidity over a similar time period. Cells from warm-acclimated, temperature down-shifted carp hyperfluidized their plasma membranes in the cold, whereas cells from cold-acclimated fish up-shifted in temperature showed no similar effect. These cells showed a complete adjustment of membrane physical state to the temperature. Total phospholipids obtained from warm-acclimated temperature down-shifted cells became more rigid than they were, when assayed at the acclimation temperature. In contrast, phospholipids obtained from cold-acclimated cells became more rigid when exposed to increasing temperatures. No significant changes occurred to the polar head groups, or to the fatty acid composition of the total phospholipids. It was concluded that the lipids play only a secondary role in the control of the physical state of plasma membrane in carp erythrocytes, and that some non-lipid components of these structures might be involved in these regulatory processes.     

Prevalence of Setaria equina microfilaraemia in horses in Hungary

Peripheral blood samples were collected randomly from 195 horses in various parts of Hungary, and the presence of microfilariae was evaluated by the Knott technique. On the basis of morphological identification 18 of the horses (9.2 per cent) were infected with Setaria equina, and the infection was confirmed in 10 animals by pcr and sequencing. The level of microfilaraemia was between 1 and 1138 larvae in 2 ml of blood. There was no correlation between the time of sampling or the sex of the animals (stallions versus mares) and the prevalence of infection, but the prevalence decreased with age. There was a significant association between the prevalence of microfilaraemia and the presence of still waters; positive samples were collected either in the region of Lake Balaton, the largest lake in the country, or at places with nearby ponds.     

Improved climate control for potato stores by fuzzy controllers

The main tasks for climate controllers in potato bulk stores are to keep the storage climate in an appropriate condition for quality conservation. To avoid extra energy costs by using cooling systems the internal climate of potato bulk stores is controlled with outdoor air. A conventionally designed controller is difficult to adapt for an optimal climate control to reduce quality loss. However, a controller designed with fuzzy logic is found as advantageous for adapting control parameters to improve the control process. A test stand is built to ventilate and to control two samples of potato bulks simultaneously with the same outdoor weather conditions for comparisons. Several experiments were carried out to compare climate controllers with different control algorithms, i.e. conventional to a different conventional and conventional to fuzzy.     

Effects of high-pressure processing at low temperature on the molecular structure and surface properties of beta-lactoglobulin

High-pressure processing (HPP) was utilized to induce unfolding of beta-lactoglobulin (beta-LG). beta-Lactoglobulin solutions at concentrations of 0.5 mg/mL, in pH 7.5 phosphate buffer, were pressure treated at 510 MPa for 10 min at either 8 or 24 degrees C. The secondary structure, as determined by circular dichroism (CD), of beta-LG processed at 8 degrees C appeared to be unchanged, whereas beta-LG processed at 24 degrees C lost alpha-helix structure. Tertiary structures for beta-LG, as determined by near-UV CD, intrinsic protein fluorescence spectroscopy, hydrophobic fluorescent probe binding, and thiol group reactivity, were changed following processing at either temperature. The largest changes to tertiary structure were observed for the samples processed at 24 degrees C. Model solutions containing the pressure-treated beta-LG showed significant decreases in surface tension at liquid-air interfaces with values of 54.00 and 51.69 mN/m for the samples treated at 24 and 8 degrees C, respectively. In comparison, the surface tension for model solutions containing the untreated control was 60.60 mN/m. Changes in protein structure during frozen and freeze-dried storage were also monitored, and some renaturation was observed for both storage conditions. Significantly, the sample pressure-treated at 8 degrees C continued to display the lowest surface tension.     

Babesia canis canis in dogs from Hungary: detection by PCR and sequencing

Canine babesiosis in Hungary has always been a severe and frequent disease, attributed to infection with Babesia canis transmitted by Dermacentor reticulatus. Identification of the disease agent has been based merely on size and morphology of the intraerythrocytic parasites and no evidence has been found concerning the subspecies (genotype) of B. canis. Therefore, a molecular survey on natural Babesia infection of dogs in Hungary using PCR and sequence analysis was attempted to clarify the subspecies (genotype) and to obtain information on the occurrence of B. canis. A total of 44 blood samples from dogs showing clinical signs of babesiosis were collected. A piroplasm-specific PCR amplifying the partial 18S rRNA gene yielded an approximately 450 bp PCR product in 39 (88.6%) samples. Thirteen positive samples originated from Budapest and 26 from 21 other locations. Five PCR products were chosen randomly for sequencing. The partial 18S rDNA sequences were submitted to GenBank (accession numbers AY611729; AY611730; AY611731; AY611732 and AY611733). The sequences showed 100% homology to one another or differed by one nucleotide. BLAST search against GenBank revealed the highest similarity (99.8 or 100%) with Babesia canis canis. The implication of these data, for the further study and diagnosis of canine babesiosis is discussed.     

Structure–activity relationships of eugenol derivatives against Aedes aegypti (Diptera: Culicidae) larvae

BACKGROUND: Dengue fever is a severe public health problem for several countries. In order to find effective larvicides to aid control programs, the structure‐activity relationships of eugenol derivatives against Aedes aegypti (Diptera: Culicidae) larvae were evaluated. Additionally, the composition and larvicidal activity of Syzygium aromaticum essential oil was assessed. RESULTS: Four compounds representing 99.05% of S. aromaticum essential oil have been identified. The essential oil was active against Ae. aegypti larvae (LC₅₀ = 62.3 and 77.0 ppm, field‐collected and Rockefeller larvae respectively). The larvicidal activity of eugenol, the major compound of the essential oil, was further evaluated (LC₅₀ = 93.3 and 71.9 ppm, field‐collected and Rockefeller larvae respectively). The larvicidal activity and structure‐activity relationships of synthetic derivatives of eugenol were also assessed. The larvicidal activity of the derivatives varied between 62.3 and 1614.9 ppm. Oxidation of eugenol allylic bond to a primary alcohol and removal of the phenolic proton resulted in decreased potency. However, oxidation of the same double bond in 1‐benzoate‐2‐methoxy‐4‐(2‐propen‐1‐yl)‐phenol resulted in increased potency. CONCLUSION: Structural characteristics were identified that may contribute to the understanding of the larvicidal activity of phenylpropanoids. The present approach may help future work in the search for larvicidal compounds. Copyright © 2012 Society of Chemical Industry     

Serological evidence for Babesia canis infection of horses and an endemic focus of B. caballi in Hungary

In order to evaluate the seroconversion of horses to Babesia caballi and B. canis in Hungary, blood samples were collected from 371 animals on 23 different locations of the country. The presence of antibodies to B. caballi was screened with a competitive ELISA. All 29 positive samples came from one region (the Hortobágy). The prevalence of infection did not show correlation with sexes, and reached 100% in the age group of 2-5 years. Babesia canis-specific antibodies were demonstrated by IFAT in 6.74% of animals kept in 7 regions. The titres were low or medium level (1:40 to 1:160), indicating that the horses had previously been exposed to this piroplasm, but their infection must have been limited. The highest seropositivity rate was observed in the age group of 3-4 years, and males (stallions and geldings) were significantly more frequently infected than females. However, neither B. caballi nor B. canis could be identified in the peripheral blood samples of infected horses by PCR. Since most of the B. caballi-positive horses remained negative in the B. canis IFAT, whereas seroconversion solely to B. canis was detected in several regions of the country, serological cross-reaction between the two species can be discounted. This is the first serological evidence of horses being naturally infected with B. canis, supporting the view that piroplasms are less host specific than previously thought.     

Pesticide diversity in rice growing areas of Northern Vietnam

Pesticide use in developing countries increases rapidly. In many regions, we miss knowledge of how frequently pesticides are applied and which active ingredients are used. We present a new cost-efficient and rapid assessment method of recording pesticides diversity in rice-dominated landscapes and present some evidence of the misuse of active ingredients in our study regions. We investigated 17 rice fields in two regions of Northern Vietnam in 2014 and 2015. At each region, we explore the abundance of pesticides used with three methods including (1) the novel approach of collecting pesticide packages close to our target rice fields, (2) observations of farmers spraying pesticides in the surrounding and (3) interviewing local farmers. By collecting pesticide packages, we found 811 packages containing 74 different active ingredients. On average, 19 active ingredients (ranging from four to 40 active ingredients) were applied with an average content of 275.3 g of active ingredients per site. Insecticide packages (39%) were most abundant followed by those of fungicides (31%), herbicides (16%) and other active ingredients (14%). On all sites, active ingredients banned in the European Union were applied by the farmers. Collecting pesticide packages proved to be an efficient and rapidly implemented method to obtain some baseline information about pesticide application (for Northern Vietnam). We suspect that improved agricultural extension services could contribute to good agricultural practices in pest management. Generally, better information and education for local farmers for appropriate use of pesticides seem a necessity.