A simple and efficient DNA isolation method for Ornithogalum L. species (Hyacinthaceae, Asparagales)

We report an efficient, simple and cost-effective protocol for the isolation of genomic DNA from Ornithogalum species. Our modification of the standard CTAB protocol includes two polyphenol adsorbents (insoluble PVPP and activated charcoal), high NaCl concentrations (4 M) for removing polysaccharides, and addition of phenol to remove proteins and other contaminants. DNA yield obtained with our protocol was 223 and 312 μg DNA g-1 of dry leaf tissue. The absorbance ratio 260/280 nm was 1.879 (O. refractum) and 1.753 (O. sibthorpii), and the absorbance ratio 260/230 nm was 1.779 (O. refractum) and 1.545 (O. sibthorpii), revealing lack of contamination. PCR amplifications of one nuclear marker (26S rDNA) indicated that this DNA isolation protocol may be used for Ornithogalum plants containing many interfering compounds for further analyses in population genetics and phylogeographic studies

Social medicine approach in the resolution of problem of contamination of public areas with dog feces and its public health relevance

Since dogs and humans share their living environments and effectuatevery close contactswith each others, with dog owners not caring adequately for their pets, there is a possibility of transmission of particular parasitic diseases from dogs to humans. In view of the fact that dogs are carriers and definitive hosts of a large number ofzoonotic endoparasites, their feces may represent a source of infection for humans and a real threat to their health. That is the reason for raising the level of awareness among citizens about the necessity of executing zoohygienic measures in dog breeding, as well as about the significance of causally planned dehelminthization and anti-ectoparasitic treatments. It is essential as well that dog owners perform regular coprological examinations, immediately after getting the dog and four times a year afterwards, abiding by the principle of shifting the preparations used based on their different chemical formulations. A timely diagnosis of intestinal parasites in dogs and proper treatment of infected animals enable the prevention of these infections in humans. Contamination of public areas with intestinal parasites from dog feces represents a public health problem that should be kept under control using the appropriate social medicine approach. Such an approach would typically involve health education measures related to the control of parasite transmission in the environment, the presentation to dog owners of the guidelines for the prevention of the occurrence, persistence, and spread of zoonotic diseases, and health education of medical and veterinary professionals, pet owners, and the wider community regarding the preservation of the health of both humans and pet animals. An increased level of knowledge and corrected attitudes and behaviors among individuals would certainly contribute to more effective prevention of zoonotic diseases

Looking for the causes of and solutions to the issue of honey bee colony losses

Colony losses, including those induced by the colony collapse disorder, are an urgent problem of contemporary apiculture which has been capturing the attention of both apiculturists and the research community. CCD is characterized by the absence of adult dead bees in the hive in which few workers and a queen remain, the ratio between the brood quantity and the number of workers is heavily disturbed in favor of the former, and more than enough food is present. Robbing behavior and pests usually attacking the weakened colony do not occur. In the present paper, the causes of the emergence of this problem are discussed, as well as the measures of its prevention. The following factors, which lead to colony losses, are analyzed: shortage of high-quality food (pollen and honey); infestation with parasites, primarily with Varroa destructor, and mixed virus infections; bacterial infections (American and European foulbrood), fungal infections (nosemosis and ascosphaerosis) and trypanosomal infections (lotmariosis); and, finally, general management of the apiary. Certain preventive measures are proposed: (1) providing ample high-quality forage and clean water, (2) avoiding sugarisation, i.e. superfluous use of sugar syrup, (3) meeting the nutritional needs of the colony, (4) when feeding bees, taking care of the timing and the composition of diet, avoiding pure sugar syrup which in excessive quantities may induce energetic and oxidative stress, (5) when there is a shortage of natural feed - honey in the brood chamber - use sugar syrup with natural/artificial supplements to avoid protein starvation, (6) organized control of V. destructor in the colonies is obligatory due to its vector role, and (7) compliance with hygienic and sanitary measures and principles of good apiculture practice and management in apiaries. To conclude, all preventive measures are feasible in compliance with rules and regulations concerning regular spring and autumn bee health monitoring by licensed veterinarians, who can propose adequate treatments if necessary

Contact varroacidal efficacy of lithium citrate and its influence on viral loads, immune parameters and oxidative stress of honey bees in a field experiment

With an almost global distribution, Varroa destuctor is the leading cause of weakening and loss of honey bee colonies. New substances are constantly being tested in order to find those that will exhibit high anti-Varroa efficacy at low doses/concentrations, without unwanted effects on bees. Lithium (Li) salts stood out as candidates based on previous research. The aims of this study were to evaluate Li citrate hydrate (Li-cit) for its contact efficacy against Varroa, but also the effect of Li-cit on honey bees by estimating loads of honey bee viruses, expression levels of immune-related genes and genes for antioxidative enzymes and oxidative stress parameters on two sampling occasions, before the treatment and after the treatment. Our experiment was performed on four groups, each consisting of seven colonies. Two groups were treated with the test compound, one receiving 5 mM and the other 10 mM of Li-cit; the third received oxalic acid treatment (OA group) and served as positive control, and the fourth was negative control (C group), treated with 50% w/v pure sucrose-water syrup. Single trickling treatment was applied in all groups. Both tested concentrations of Li-cit, 5 and 10 mM, expressed high varroacidal efficacy, 96.85% and 96.80%, respectively. Load of Chronic Bee Paralysis Virus significantly decreased (p < 0.01) after the treatment in group treated with 5 mM of Li-cit. In OA group, loads of Acute Bee Paralysis Virus and Deformed Wing Virus significantly (p < 0.05) increased, and in C group, loads of all viruses significantly (p < 0.01 or p < 0.001) increased. Transcript levels of genes for abaecin, apidaecin, defensin and vitellogenin were significantly higher (p < 0.05—p < 0.001), while all oxidative stress parameters were significantly lower (p < 0.05—p < 0.001) after the treatment in both groups treated with Li-cit. All presented results along with easy application indicate benefits of topical Li-cit treatment and complete the mosaic of evidence on the advantages of this salt in the control of Varroa

Experimental investigation of spillover effect in system of active vibration control

Piezoelektrični aktuatori se uveliko primenjuju za aktivno upravljanje vibracijama na raznim strukturama sa ciljem povećanja performansi sistema. Razvijeni sistem aktivnog upravljanja vibracijama se sastoji od aktivnije strukture, kontrolera i visokonaponskih pojačivača. U ovom radu, kompozitna greda predstavlja osnovnu strukturu za senzorsku (merne trake) i aktuatorsku (dvoslojni PZT piezoelektrični aktuator) platformu. U cilju unapređenja dinamičkih karakteristika aktivnog sistema vršena je promena koeficijenata PID regulatora. Efikasnost sistema aktivnog upravljanja vibracijama za posmatrani mod oscilovanja se može povećati promenom koeficijenata PID regulatora, ali u isto vreme moguće je narušiti stabilnost sistema. Nestabilnost sistema je najčešće uslovljena pojavom 'spillover' efekta. U ovom radu predstavljen je značaj 'spillover' efekta u zatvorenim petljama piezoelektričnih aktivnih struktura, kao i značaj promene koeficijenata PID regulatora na stabilnost sistema. Eksperimentalni rezultati sistema aktivnog prigušenja vibracija su predstavljeni u radu i daju jednu od mogućnosti za povećanje stabilnosti upravljanih struktura.Piezoelectric actuators are widely used in structural systems for active vibration control with the aim to enhance the performance of systems. The developed system of active vibration control consists of active structure, controller and high voltage amplifier. In this paper, the composite beam is host structure for sensor platform (strain gages) and actuator platform (dual layer PZT piezoelectric actuator). In order to improve the dynamic characteristics of active system, the coefficients of PID controller are changed. The effectiveness of active vibration control system at mode of interest can be improved with change of PID coefficients, but the stability of system can be reduced. The instability of the active structure is often perturbed by spillover effect. In this paper the importance of considering spillover effects in closed loop of piezoelectric active structures is demonstrated and shown the importance of change the PID coefficients in stability of active vibration control system. Experimental results which correspond to the developed active vibration control system are presented and affirmed stability on proposed active structure

Experimental investigation of spillover effect in system of active vibration control

Piezoelektrični aktuatori se uveliko primenjuju za aktivno upravljanje vibracijama na raznim strukturama sa ciljem povećanja performansi sistema. Razvijeni sistem aktivnog upravljanja vibracijama se sastoji od aktivnije strukture, kontrolera i visokonaponskih pojačivača. U ovom radu, kompozitna greda predstavlja osnovnu strukturu za senzorsku (merne trake) i aktuatorsku (dvoslojni PZT piezoelektrični aktuator) platformu. U cilju unapređenja dinamičkih karakteristika aktivnog sistema vršena je promena koeficijenata PID regulatora. Efikasnost sistema aktivnog upravljanja vibracijama za posmatrani mod oscilovanja se može povećati promenom koeficijenata PID regulatora, ali u isto vreme moguće je narušiti stabilnost sistema. Nestabilnost sistema je najčešće uslovljena pojavom 'spillover' efekta. U ovom radu predstavljen je značaj 'spillover' efekta u zatvorenim petljama piezoelektričnih aktivnih struktura, kao i značaj promene koeficijenata PID regulatora na stabilnost sistema. Eksperimentalni rezultati sistema aktivnog prigušenja vibracija su predstavljeni u radu i daju jednu od mogućnosti za povećanje stabilnosti upravljanih struktura.Piezoelectric actuators are widely used in structural systems for active vibration control with the aim to enhance the performance of systems. The developed system of active vibration control consists of active structure, controller and high voltage amplifier. In this paper, the composite beam is host structure for sensor platform (strain gages) and actuator platform (dual layer PZT piezoelectric actuator). In order to improve the dynamic characteristics of active system, the coefficients of PID controller are changed. The effectiveness of active vibration control system at mode of interest can be improved with change of PID coefficients, but the stability of system can be reduced. The instability of the active structure is often perturbed by spillover effect. In this paper the importance of considering spillover effects in closed loop of piezoelectric active structures is demonstrated and shown the importance of change the PID coefficients in stability of active vibration control system. Experimental results which correspond to the developed active vibration control system are presented and affirmed stability on proposed active structure

EPIDEMIOLOGICAL AND CLINICAL IMPORTANCE OF CANINE DIOCTOPHIMOSIS

Dioctophimosis is an endoparasitism in dogs caused by Dioctophyma renale, a nematode with an indirect life cycle and high zoonotic potential. Infection in dogs occurs through ingestion of transitional/paratenic hosts containing infective larvae. The preferred site of parasitism is the right kidney, although the parasite may also be ectopically localized in subcutaneous tissue, internal organs, peritoneum, and mediastinum. Since dogs are usually infected with a small number of parasites, the disease is asymptomatic, making the diagnosis of this disease in clinical veterinary practice rather difficult. In addition, as dogs are usually infected with a small number of parasites, the disease is asymptomatic, which makes the diagnosing of this disease in clinical veterinary practice difficult. The long prepatent period, lasting three to five months, makes it impossible to diagnose the disease in dogs younger than six months. Clinical suspicion of dioctophimosis in dogs is established on the basis of anamnestic/epizootiologic data and the clinical picture, while the final diagnosis is made ante mortem or post mortem on the basis of reliable diagnostic parameters and methods. The most practical application in routine diagnosis of canine dioctophimosis is the detection of D. renale eggs in urine sediment. When dioctophimosis is suspected in carnivores, the differential diagnosis should exclude urinary capillariosis, feline polycystic nephropathy, leptospirosis, renal fibrosis, and nephritis of various etiology. The outcome of treatment is uncertain and includes surgical removal of the affected kidney or drug therapy with avermectin derivatives. Prophylaxis consists of preventing contact of dogs with potential transitional hosts and controlling the feeding of their heat-untreated meat. Because of the potential risks that D. renale may have on the health of dog owners, education on the etiopathogenesis and means of occurrence, maintenance, and spread of this nematode is necessary

Identification of seed coat phenolic compounds from differently colored pea varieties and characterization of their antioxidant activity

The phenolic composition of seed coats in four differently colored pea varieties (Pisum sativum L.) was investigated using UHPLC-LTQ OrbiTrap MS. The obtained findings revealed that the seed coats of the examined pea genotypes possess a unique phenolic composition compared to previously studied European cultivars. In total, 41 phenolic compounds have been identified. The seed coats of the studied cultivars contained certain amounts of rosmarinic acid, rutin, galangin, morin, naringin, hesperetin and pinocembrin as well as ten flavonol glycosides that had not been reported previously. Additionally, the total phenolic content, antioxidant activity and metal chelating capacity of extracts was determined using Folin-Ciocalteu's method, 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, ferric ion-reducing capacity and ferrous ion-chelating capacity assay, respectively. Dark colored genotypes MBK 168 and MBK 173 possessed the highest total phenolic contents as well the strongest antioxidant activities. On the other hand, bright colored genotypes MBK 88 and MBK 90 exhibited the strongest metal-chelating capacities. The examined pea seed coats may be considered as important potential contributors to human health due to the presence of bioactive phenolic constituents. In addition, our results could be used as a guideline for breeding new pea cultivars with high antioxidant activities applicable in the formulation of functional food products

Potential of wormwood and oak bark-based supplement in health improvement of Nosema ceranae-infected honey bees

Nosema ceranae, a microsporidian parasite, as one of the stressors that contribute to honey bee decline, has a significant negative impact on the longevity, productivity, and reproductive capacity of honey bee colonies. There are several different strategies for Nosema infection control, including natural-based and antibiotic-based products. In this study, we tested wormwood and oak bark-based supplement “Medenko forte” on survival, Nosema infection, oxidative stress, and expression of immune-related genes in artificially N. ceranae-infected bees. The results revealed a positive influence on the survival of Nosema-infected bees, irrespectively of the moment of supplement application (day 1, day 3, or day 6 after bee emergence), as well as reduction of Nosema loads and, consequently, Nosema-induced oxidative stress. Supplementation had no negative effects on bee immunity, but better anti-Nosema than immune-stimulating effects were affirmed based on expression levels of abaecin, defensin, hymenoptaecin, apidaecin, and vitellogenin genes. In conclusion, the tested supplement „Medenko forte” has great potential in the health protection of Nosema-infected bees. However, further investigations need to be performed to elucidate its mechanisms of action

Effects of plant-based supplement on oxidative stress of honey bees (Apis mellifera) infected with Nosema ceranae

One of the most important approaches in the prevention and treatment of nosemosis is the use of herbal preparations as food supplements for bees. Therefore, the aim of this study was to investigate the effects of a plant-based supplement branded as “B+” on honeybees in a laboratory experiment. Four experimental groups were established: treated group (T), N. ceranae-infected and treated group (IT), N. ceranae-infected group (I) and non-infected group (NI). Survival, N. ceranae spore load and oxidative stress parameters together with expression levels of antioxidant enzyme genes and vitellogenin gene were monitored. The mortality in the T, IT and NI groups was significantly (p < 0.001) lower than in than in the I group. Within Nosema-infected groups, the IT group had a significantly lower (p < 0.001) number of N. ceranae spores than the I group. In addition, expression levels of genes for antioxidant enzymes were lower (p < 0.001) in the IT group compared to the I group. The concentration of malondialdehyde and the activities of antioxidant enzymes (superoxide dismutase, catalase and glutathione S-transferase) were significantly lower (p < 0.001) in the IT group compared to the I group. No negative effects of the tested supplement were observed. All these findings indicate that the tested supplement exerted beneficial effects manifested in better bee survival, reduced N. ceranae spore number and reduced oxidative stress of bees (lower expression of genes for antioxidant enzymes and oxidative stress parameters)