Cover Image,Volume 75, Issue 3

The front cover image is based on the Research Article Does winter supplementary feeding affect deer damage in a forest ecosystem? A field test in areas with different levels of deer pressure by Zbigniew Borowski et al., DOI: 10.1002/ps.5131 . Image Credit: Karol Zub.

     

In vitro effects of beta-hydroxy-beta-methylbutyrate (HMB) on cell-mediated immunity in fish

beta-Hydroxy-beta-methyl butyrate(HMB) has been shown to counteract many of the negative effects of intensive animal production methods and results in increased growth and protection against diseases. In the present study, the effect of HMB on the immunocompetence cell activity in rainbow trout (Oncorhynchus mykiss) and carp (Cyprinus carpio) was examined. Pronephric phagocytes and lymphocytes were isolated from the fish and grown in culture medium (RPMI-1640) containing either 0, 0.1, 1, 5, 10, 25, 50 or 100 microg HMB/ml of medium. The effects of HMB on the respiratory burst activity (RBA) stimulated by phorbol myristate acetate (PMA), the potential killing activity (PKA) and lymphocyte proliferation stimulated by either concanavalin A (Con-A) or lipopolysaccharide (LPS) were examined. The addition of HMB to the culture medium increased the RBA by up to 84% (p<0.01) over that of cells grown without HMB. Similarly, the PKA of the phagocytes was also increased with HMB addition to the medium by up to 140% (p<0.01) over that of cells grown without HMB. Lymphocyte proliferation stimulated by both ConA and LPS was also increased approximately two-fold (p<0.01) when HMB was added to the culture medium at concentrations between 10 and 100 microg HMB/ml in both rainbow trout and carp. The greatest effects of HMB on RBA and PKA activities were observed at a concentration >50 microg HMB/ml while lymphocyte proliferation was maximally stimulated at 25 microg HMB/ml. In conclusion, the current study shows that HMB could potentially improve immunocompetence cell activity in fish through increased cell proliferation and functionality.     

An antisense coat protein gene confers immunity to potato leafroll virus in a genetically engineered potato

The Bzura commercial potato cultivar was transformed by sense or antisense constructs which included the coat protein gene of potato leafroll virus RNA. In the sense construct, the coat protein gene was preceded by a leader sequence shorter than that in the subgenomic RNA formed in infected cells. The antisense construct consisted of a sequence complementary to the first 2020 nucleotides of the subgenomic RNA. Selected transformants expressing viral RNA were resistant to virus challenge by viruliferous aphids. In one line, expression of the antisense RNA prevented virus infection even after grafting with scions from infected plants and therefore this transformant might be regarded as virus immune.     

Genetic diversity of apricot revealed by a set of SSR markers from linkage group G1

Eight polymorphic simple sequence repeat (SSR) markers located in the G1 linkage group of apricot (Prunus armeniaca L.) were previously developed and evaluated in a small set of cultivars. Those primers were used for studying variability in 77 apricot cultivars belonging to five different geographical groups, such as Chinese, Asian (Irano-Caucasian and Central Asian), North American, Mediterranean and Western European as well as Middle European cultivars. Six of the markers were polymorphic and revealed a total of 71 alleles ranging from 5 (aprigms11) to 20 (aprigms1) alleles per locus with a mean value of 11.83 alleles per locus. In conclusion, the SSR loci located in the G1 linkage group show a level of polymorphism which is similar to loci dispersed throughout the entire genome. The total number of alleles and the number of unique alleles were the highest in Chinese apricots and the lowest in Middle European cultivars. Heterozygosity also showed a decrease from Asia and China to Middle Europe. No association could have been observed between any SSR markers tested and plum pox virus (PPV) resistant phenotype of cultivars. PPV resistant cultivars did not form a separate clade on the dendrogram obtained by UPGMA cluster analysis. Middle European and Chinese cultivars formed separate clusters while other genotypes formed smaller multiple sub-groups or scattered among different clusters. Our results support previous hypotheses on the origin of PPV resistance in North American apricots. The allele data was also presented in a form that allowed the easy observation of allele frequencies in each geographical group at each locus. Using this data field, differences and similarities between cultivar groups can be easily assessed. The analysis demonstrated the links between the North American and Mediterranean apricot germplasm and confirmed that the Chinese and Eastern European cultivars are distantly related.     

Reproduction of chub, Leuciscus cephalus L., under controlled conditions

Research into artificial reproduction of chub was studied during three subsequent reproductive seasons. Chub reproduction was conducted in two different variations. In the first experiment different hormonal agents were given to fish, such as CPE, hCG, or an analogue of GnRH with dopamine antagonist (ovopel). In the second one, the optimum moment of applying of artificial reproduction was sought and the fish were only obtained an analogue of GnRH in different periods. The results obtained in two first seasons indicated that ovopel is the most effective agent for the artificial reproduction of chub. Consequently, it was decided that the fish would obtain only this agent in the final year of research. In last year (experiment II), chub spawners were taken to the hatchery under controlled conditions every few weeks from February to the beginning of June. Periodically, the maturity of oocytes was checked. As soon as the oocytes had achieved stage 2–3 of maturity, fish were obtained ovopel. It was noted that there was no problem with collecting semen. Obtaining oocytes was much more difficult. Most frequently, they were not obtained at all or obtained in very small amounts. It was also observed that the administration of hormones caused a very quick maturation of gonads and their over-maturation.     

Seminal plasma proteins of Atlantic halibut (Hippoglossus hippoglossus L.)

Protein content and properties in the seminal plasma of Atlantic halibut (Hippoglossus hippoglossus) were assayed using spectrophotometric and electrophoretic methods. The protein concentration ranged from 6.4 ± 3.1 to 19.4 ± 3.4 mg ml⁻¹ and anti-proteolytic activity from 585.2 ± 104.6 to 2912.4 ± 367.4 U l^−l. A high correlation between anti-proteolytic activity and protein concentration (r = 0.95), and between sperm concentration and osmolality was found (r = 0.92). There was a significant decrease in anti-proteolytic activity from the first to the second sampling, but not in protein concentration. Anti-proteolytic activity and protein concentration were significantly affected by variations in individual males. Electrophoresis revealed four anti-proteolytic bands and individual differences in bands of proteolytic activity, which were subsequently characterized as metalloproteases and serine proteases.     

Loss in macronutrient pools in bilberry and lingonberry in mesic Scots pine forests after Northern red oak introduction

Forest transformation from coniferous monocultures to mixed stands is being promoted worldwide, including the introduction of fast-growing broadleaved tree species within native stands. Here, we studied how enrichment of temperate European Scots pine (Pinus sylvestris) forest by North-American northern red oak Quercus rubra impacted macronutrient concentrations in two long-lived and dominant components of the forest understory: bilberry Vaccinium myrtillus and lingonberry V. vitis-idaea. Study sites were located in forest complexes (central Poland) which occupy continuously reforested lands (hereafter ancient forests) as well as post-agricultural lands (recent forests), all suitable for mesic pine forests. Samples of bilberry and lingonberry leaves, stems, and fruits were collected in pine stands and in adjacent Scots pine-red oak stands, in both ancient and recent forests. Concentrations of macronutrients (C, N, P, K, Ca, S, and Mg) in aboveground biomass components were analysed using standardized chemical procedures. The study revealed intra- and interspecific (bilberry vs. lingonberry) differences in concentrations of all nutrients in leaves, stems, and fruits, except for invariable C concentrations. Macronutrient accumulations in plants were decreased by land-use discontinuity and favoured by enrichment of tree stands by Q. rubra. The estimated macronutrient pools were much higher for V. myrtillus than V. vitis-idaea in all forest types studied. They were lower in forests enriched with Q. rubra, both ancient (up to 25.5% for bilberry and 99.9% for lingonberry) and recent (46.9% and 99.9%, respectively), as well as in recent pine forest (46.6% and 81.1%, respectively) than in ancient pine forest. Higher K and S pools (39.3% and 6.5%, respectively) noted for bilberry in an ancient forest with Q. rubra were exceptions. Despite more effective accumulations of elements at the species level, macronutrient pools of Vaccinium myrtillus and V. vitis-idaea decreased significantly in the presence of introduced Q. rubra due to negative impacts of this broadleaved tree on bilberry and lingonberry cover and biomass. Therefore, the limitation of alien Q. rubra planting in sites of mesic pine forest with the abundant occurrence of V. myrtillus and/or V. vitis-idaea is recommended.

Graphic abstract

     

Rates of oxygen consumption and ammonia excretion of juvenile Eurasian perch Perca fluviatilis L.

The impact of feeding, fish size (body weight from 18.5 to 56.5 g) and water temperature (20 and 23 °C) on oxygen consumption (OC, mg O₂ kg^–1 h^–1) and ammonia excretion (AE, mg TAN kg^–1 h^–1) was studied in Eurasian perch held in recirculation systems. OC for both fed and feed-deprived (3 days) fish was higher at 23 °C (278.5 and 150.1 mg O₂ kg^–1 h^–1) than at 20 °C (249.3 and 135.0 mg O₂ kg^–1 h^–1; P < 0.01). AEs for both fed and feed-deprived fish were also significantly higher at 23 °C than at 20 °C (P < 0.001). Water temperature and fish size had a significant impact on the oxygen:feed ratio (OFR, kg O₂ kg^–1 feed fed day^–1) and ammonia:feed ratio (AFR, kg TAN kg^–1 feed fed day^–1; P < 0.001). Their average values at temperatures of 20 and 23 °C were 0.17 and 0.19 kg O₂ kg^–1 feed fed day^–1 and 0.009 and 0.011 kg TAN kg^–1 feed fed day^–1, respectively.     

Normal electrocardiographic and echocardiographic (M-mode and two-dimensional) values in Polish Landrace pigs

Background

Swine are recognized animal models of human cardiovascular diseases. Normal values of cardiac morphology and function have been published for swine but for smaller number of pigs and not for swine whose weights ranged up 100 kg. In order to improve the value of results of an investigation on cardiac morphology and function in swine when such data are extrapolated to humans, the aim of this study was to document electrocardiographic and echocardiographic measures of cardiac morphology and function in swine. The study comprised 170 single and repeated measurements that were made in 132 healthy domestic swine (Sus domesticus) whose weights ranged between 20-160 kg and were used as controls in three different experiments. All electrocardiographic and echocardiographic measurements in all swine were done under general anaesthesia.

Results

Statistically significant correlations were found between body weight and heart rate (HR), the duration of the P-wave, the duration of the QRS interval, the duration of the QT interval, and the corrected QT ratio (QTc). Since body weight was positively correlated with age, statistically significant correlations were also found between age and HR, the duration of the P-wave, the duration of the QRS interval, the duration of the QT interval, and the QTc. We found that the thickness of the left ventricular wall and the internal diameter of the left ventricle increased with age and body weight. We also found positive trends between body weight and ejection fraction and body weight and fractional shortening. We also found a positive relationship between age, body weight, and the ratio of the left ventricular internal diameter to its wall thickness, as well as the relative left atrial size.

Conclusion

Many electro- and echocardiographic measures of cardiac morphology and function of healthy swine are related to their body weight. When the electro- and echocardiographic measures of domestic swine and humans are compared, the most comparable electrocardiographic values are those that were determined in swine whose body weights are not greater than 70 kg. In contrast, the most comparable echocardiographic measures are those that were determined in swine with a body weight of 40–110 kg.     

Influence of β-hydroxy-β-methylbutyrate on nonspecific humoral defense mechanisms and protection against furunculosis in pikeperch (Sander lucioperca)

Studies have shown that in both in vitro and in vivo tests, β‐hydroxy‐β‐methylbutyrate (HMB) increases the nonspecific cellular and humoral immune response and protection against diseases in animals. The present study examines the influence of HMB on nonspecific humoral defense mechanisms and protection against furunculosis in pikeperch (Sander lucioperca). β‐hydroxy‐β‐methylbutyrate was fed in a pelleted ration of 50 mg kg^?1 feed day^?1 for 4 weeks. Blood was drawn from 12 HMB‐fed and control‐fed pikeperch. The lysozyme and ceruloplasmin activities in the plasma, total immunoglobulin (Ig) levels, and total serum protein were analysed prior to and then after 2 and 4 weeks of HMB ingestion. After 4 weeks of HMB ingestion, a challenge test was performed by injecting the fish with live pathogenic Aeromonas salmonicida bacteria. β‐hydroxy‐β‐methylbutyrate at a dose of 50 mg kg^?1 feed resulted in a statistically significant (P<0.05) increase in the lysozyme activity of the plasma, total Ig, and serum protein levels. Additionally, reduced mortality (40%) after the in vivo challenge with pathogenic A. salmonicida suggested that HMB‐activated nonspecific protection against furunculosis in pikeperch.