Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice

Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis.     

Purification of chinook salmon (Oncorhynchus tshawytscha) GH for receptor study

A method for the purification of chinook Salmon (Oncorhynchus tshawytscha) GH, which retains its biological activity, is described. The biological activity was investigated with an established radioreceptor assay using liver membranes from pregnant rabbits and bovine GH as standard and labelled hormone. The enrichment of the preparation was checked with electrophoresis (SDS-PAGE). Extraction and further steps were carried out using low molarity alkaline buffer (pH 8–10, M = 100 mM). Three chromatography steps were performed (Concanavalin-A sepharose, Bio-gel P60, DEAE). Ion exchange chromatography was performed under isocratic conditions (using a 50 cm column). Two isoforms (sGH1 and sGH2) were isolated. The purification yield is 0.7% compared to lyophilized pituitaries. The molecule is homogeneous in SDS-PAGE. Contamination by prolactin, gonadotrophin and corticotrophin is negligible (< 0.5%). It could be demonstrated that the biological activity of the preparation is maintained since this preparation stimulates the growth of juvenile trout (Salmo gairdneri) and binds specifically (35%) to trout liver membranes.     

Prophylactic and therapeutic application of Propionibacterium avidum KP-40 in swine and calves with acute enzootic bronchopneumonia.

The usefulness of the prophylactic or therapeutic application of Propionibacterium avidum KP-40, a potent stimulator of the monocyte-macrophage-system, was demonstrated in piglets and calves. After a 3-month-period of observation PA-treated piglets showed a significantly improved development (decreased number of infections, gain of body weight). In piglets and calves the therapeutic use of PA together with oxytetracycline proved to be superior in the treatment of acute endemic enzootic bronchopneumonia (AEB) as compared to groups of animals receiving PA or oxytetracycline alone.     

Protection of Groundwater from Migration of Infiltrates from a Chromic Waste Storage Site and Methods of Treating these Infiltrates

This work presents the results ofinvestigations to develop and implement methods toeffectively collect and purify infiltrates from heaps,situated in the region of Alwernia near Cracow, wheremore than 3 million tonnes of waste material resultingfrom the production of chromium compounds have beenstored. It describes a system for the protection ofgroundwater from these infiltrates which contain 50–400 g m^-3 Cr⁶⁺, as well as the effectivenessof cheap and simple chemical methods to purify thesechromic wastewaters. The infiltrate collection systemand the most effective method to decrease theconcentration of Cr⁶⁺ to a level below 0.1 ppm,as required by Polish and European Union regulations,were implemented in the Alwernia Chemical Works S.A.in the years 1998–1999.     

Influence of long-term vaccination of a breeding herd of pigs against PCV2 on reproductive parameters

The objective of the study was to evaluate an efficacy of sows vaccination protocols in the herd with serious problems affecting efficacy of reproduction. The study was performed in a large pig herd with about 1200 sows. Before vaccination against PCV2, farrowing rate in this farm was about 65%. Sows, boar and replacement gilts were immunized using Circovac vaccine (Merial, France) according to producer's recommendations. Parameters of production were analyzed since 2007 until 2010 in selected batches of sows inseminated at the same weeks of the year (17th, 18th, 19th and 20th) to eliminate seasonal variability. In total, 940 sows were subjected to the study. No significant changes in management during these years were introduced. The applied protocol of sow herd long-term vaccination proved to be very efficient. All measured production parameters: reproduction rate, number of piglets born alive, birth weight of piglets and number of piglets weaned per a litter improved after implementation of immunization program. Moreover, further improvement was observed with vaccination in the following reproduction cycles. The most spectacular effect of vaccination regarded average farrowing rate that increased from 64.76% in control group to 86.93% after basic vaccination. Two years after implementation of vaccination program this parameter reached 93.6%. Number of piglets weaned per sow per a litter improved from 10.31 to 11.74 after one year of vaccination and remained relatively stable through the following year. Simultaneously, the percentage of newborn piglets with birth weight < 1 kg decreased significantly (p < 0.05). To summarize, vaccination against PCV2 influenced positively the insemination rate, number of piglets born alive and weaned per litter as well as birth body weight and percentage of piglets weighing < 1 kg.     

Antimicrobial susceptibility of Brachyspira hyodysenteriae isolated from 21 Polish farms

Swine dysentery (SD) is a common disease among pigs worldwide, which contributes to major production losses. Antimicrobial susceptibility testing of B. hyodysenteriae, the etiological agent of SD, is mainly performed by the agar dilution method. This method has certain limitations due to difficulties in interpretation of results. The aim of this study was the analysis of antimicrobial susceptibility of Brachyspira hyodysenteriae (B. hyodysenteriae) Polish field isolates by broth microdilution procedure. The study was performed on 21 isolates of B. hyodysenteriae, collected between January 2006 to December 2010 from cases of swine dysentery. VetMIC Brachyspira panels with antimicrobial agents (tiamulin, valnemulin, doxycycline, lincomycin, tylosin and ampicillin) were used for susceptibility testing of B. hyodysenteriae. The minimal inhibitory concentration (MIC) was determined by the broth dilution procedure. The lowest antimicrobial activity was demonstrated for tylosin and lincomycin, with inhibition of bacterial growth using concentrations > 128 microg/ml and 32 microg/ml, respectively. In the case of doxycycline, the MIC values were < or = 2.0 microg/ml. No decreased susceptibility to tiamulin was found among the Polish isolates and MIC values for this antibiotic did not exceed 1.0 microg/ml. The results of the present study confirmed that Polish B. hyodysenteriae isolates were susceptible to the main antibiotics (tiamulin and valnemulin) used in treatment of swine dysentery. Further studies are necessary to evaluate a possible slow decrease in susceptibility to tiamulin and valnemulin of B. hyodysenteriae strains in Poland.     

Tulathromycin enhances humoral but not cellular immune response in pigs vaccinated against swine influenza

The effect of a standard, single dose therapy with tulathromycin was investigated on the postvaccinal humoral and cellular immune response in pigs vaccinated against swine influenza. Forty‐five pigs, divided into 3 groups, were used (control not vaccinated (C, n = 15), control vaccinated (CV, n = 15), and experimentally received tulathromycin (TUL, n = 15)). For vaccination of pigs, an inactivated, commercial vaccine was used. Pigs from TUL group received single dose of tulathromycin intramuscularly, at the recommended dose (2.5 mg/kg body weight). Pigs from TUL and CV groups were vaccinated at 8 and 10 weeks of age. The specific humoral and cellular immune response against swine influenza virus (SIV) was evaluated. The results of present study showed that humoral postvaccinal response after vaccination against SIV can be modulated by treatment with tulathromycin. In pigs from TUL group, the significantly higher titers of anti‐SIV‐specific antibodies were observed 4 and 6 weeks after booster dose of vaccine. Simultaneously, T‐cell‐mediated immune response against SIV was not affected by tulathromycin. Our recent study confirmed the importance of defining the modulatory activity of tulathromycin because of its influence on the immune response to vaccines. Since the antibodies against hemagglutinin are crucial for the protection against SIV, the present observations should prompt further studies on the practical significance of recent results in terms of clinical implications (postvaccinal protection) in the field conditions.     

Wear of spring tine cultivator points in sandy loam and light clay soils in southern Poland

Correct evaluation and forecasting of durability of soil cutting parts are the decisive factors for proper operation of agricultural machinery. During operation of cultivators, over two-fold difference in wear of cultivator points is observed, which makes establishing of their exchange periods extremely difficult. So, there is a need to conduct some research aimed for determination of relationship between arrangement of the points in individual cultivator rows and their wear. The presented experiments were performed under authentic farm conditions during spring soil loosening and levelling operations before sowing and planting, in sandy loam and light clay soils. Changes in length and thickness of the spring tine points, wear patterns and weight loss were measured. Loss in length of spring tine points was described by a complex function of the distance travelled or by an exponential function of the distance from the measuring point to the symmetry axis of a spring tine point; loss in thickness was described by a linear function. It was found that soil type, as well as arrangement of the points in individual cultivator rows and their position in relation to the tractor wheel track, significantly influenced their wear. It was also observed that wear of spring tine points was 40–100% higher in the sandy loam soil than in the light clay soil, and that wear within a tractor wheel track was 17–40% higher than outside the track. Moreover, wear of points positioned in the first row on the cultivator frame was 26–100% higher than wear of those in the third row. The research results emphasise the need for field experiments at compare wear of spring tine points made of different materials or of different designs to take into account differences in wear due to positioning of the points on cultivator frames.     

Effect of maternal antibodies and pig age on the antibody response after vaccination against Glässers disease

The influence of age and maternal antibodies on the development and duration of postvaccinal antibody response against Glässer’s disease were investigated. Pigs born to immune (MDA-positive) and non-immune (MDA-negative) sows were vaccinated with inactivated vaccine. Vaccination was done according to three different protocols: at 1 and 4, at 2 and 5 or at 4 and 7 weeks of age. There were also two control groups for MDA-negative and MDA-positive pigs. The level of Haemophilus parasuis (Hps) specific antibodies were determined using commercial ELISA test. No serological responses were seen in any of the groups after the first vaccination. Maternally derived antibodies (MDA) against Hps were above the positive level until approximately 3 weeks of life in MDA-positive pigs. In those pigs the strongest postvaccinal humoral response was observed in piglets vaccinated at 4 and 7 weeks of age. In the remaining MDA-positive piglets only slight seroconversion was noted but levels of antibodies never exceeded values considered as positive. All MDA-negative pigs produced Hps-specific antibodies after the second vaccination. The results of the present study indicated that MDA may alter the development and duration of active postvaccinal antibody response. Age of pigs at the moment of vaccination was not associated with the significant differences in the magnitude of antibody response, however influenced the kinetics of decline of Hps-specific antibodies.     

Application of image analysis for the varietal classification of barley:: Morphological features

This paper presents an exploratory investigation of the application of morphological features in image analysis for varietal classification of Polish spring barley. The objective of this study was to determine the utility of morphological features for classifying individual kernels of five varieties of barley. Furthermore, this study was performed to find the best method to classify kernels of barley with the lowest error of classification. Image processing consisted of several steps: image acquisition, segmentation, external and internal image feature extraction, classification and interpretation. Each barley kernel was described using 74 morphological features. The selection was carried out using three methods based on: Fisher's coefficient, probability of error and average correlation coefficient and mutual information. Principle component analysis (PCA), linear discriminant analysis (LDA), and non-linear discriminant analysis (NDA) were used throughout this paper as the classification methods. The results confirmed that the method using morphological features may be successfully employed in image analysis for preliminary varietal identification of barley kernels. Furthermore, LDA was found to be the method which best separated different varieties of objects.