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Protozoa of the genus Giardia cause the disease giardiosis in a large number of animal species, but also in humans. This disease can be either subclinical or followed by diarrhea, malabsorption, weight loss and stunted growth. The aim of this study was to examine the presence of Giardia sp. in different species of animals in Belgrade Zoo (asymptomatic infection) and to compare the sensitivity of parasitological and molecular-genetic methods. Examination of the presence of Giardia sp. protozoa was performed in 52 fecal samples, using fecal flotation and molecular-genetic PCR method. No positive samples were detected when deploying the parasitological method, while PRC analysis revealed the presence of Giardia sp. in one animal - ring-tailed lemur (Lemur catta), which indicates the advantage of the molecular-genetic method over the parasitological one. The absence of Giardia sp. in other samples can be explained by adequate housing conditions, successful implementation of hygienic and sanitary measures, as well as good veterinary practice. Use of PCR method for testing the presence of Giardia sp. showed a higher sensitivity when compared to the fecal flotation method.

Nosema ceranae, a microsporidium species, is among the most common causes of bee diseases. The positive effect of Agaricus bisporus mushroom extract on the survival and immunity of Nosema-infected bees has been reported recently. The effect could be achieved by stimulating the expression of immune-related genes, but also by suppressing nosemosis. The aim of this work was to determine the effect of A. bisporus extract on the oxidative status of bees infected with N. ceranae. In a cage experiment on newly hatched bees, the effect of aqueous extract of champignon (A. bisporus, strain A15) was investigated. Six groups were formed: three groups were infected and received A. bisporus extract through food at different times (days 1, 3, and 6 after hatching), one group received the extract but was not infected (treatment control), one was only infected with Nosema (positive control) and one was neither infected nor received the extract (negative control). The effects were examined on samples taken on days 7 and 15 of the study. The activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione S-transferase (GST) and the concentrations of malondialdehyde (MDA) were determined. In comparison to the positive control, the enzyme activities and MDA concentrations were significantly lower in the groups fed with the mushroom extract supplement. In the negative control, the level of oxidative stress was lower than in the positive control. In comparison with the other groups, the values mostly did not differ significantly. The oxidative status of bees infected with N. ceranae was significantly better if they were fed with the A. bisporus extract.

The hygienic behavior in honey bees is a complex polygenic trait that serves as a natural defense mechanism against bacterial and fungal brood diseases and Varroa destructor mites infesting brood cells. The aim of this study was to evaluate the effect of a dietary amino acids and vitamins supplement “BEEWELL AminoPlus” on hygienic behavior of Apis mellifera colonies combating microsporidial and viral infections. The experiment was performed during a one-year period on 40 colonies alloted to five groups: one supplemented and infected with Nosema ceranae and four viruses (Deformed wing virus - DWV, Acute bee paralysis virus - ABPV, Chronic bee paralysis virus - CBPV and Sacbrood virus – SBV), three not supplemented, but infected with N. ceranae and/ or viruses, and one negative control group. Beside the l isted pathogens, honey bee trypanosomatids were also monitored in all groups.

Following the “one health” principle, we have conducted optimization of a protocol for β-casein genotyping in cattle in order to select cows with exclusively the A2A2 genotype. Gastrointestinal proteolysis of A1 β-casein in humans releases beta-casomorphin 7, which is believed to cause a number of diseases/conditions (diabetes mellitus type 1, ischemic heart disease, atherosclerosis, sudden infant death syndrome, autism, schizophrenia, gastrointestinal discomfort, and prolonged gastrointestinal passage time). On the contrary, A2 β-casein does not cause similar effects on human health, due to its different metabolism. DNA extraction was conducted from blood samples belonging to the laboratory archive of the Department of Biology, Faculty of Veterinary Medicine, University of Belgrade. Determination of genotypes was performed using the Allele Specific Polymerase Chain Reaction (AS-PCR) method. The amplification was preceded by determination of proper primer annealing temperature (65.50 °C), in order to ensure optimal genotyping results. The results obtained indicated a higher frequency of the A2 allele (0.56) compared to the A1 allele (0.44). Furthermore, in 7 out of 35 tested samples, the A1A1 genotype (20.00%) was found, in 17 samples, the A1A2 genotype (48.60%) was found, and in 11 samples, the A2A2 genotype (31.40%) was found. The molecular methods used ensured reliable β-casein genotyping that would enable selection of cows with the A2A2 β-casein genotype, implying production of.

Diabetes is one of the main health concerns, especially in developed countries. During the last few decades, the percentage of diabetic persons is constantly increasing. Although the genetic factors have a strong influence in the development of diabetes, environmental influence (physical inactivity, inadequate nutrition leading to obesity) also have an important impact. The main objective of this investigation was to evaluate the possible influence of organic sprouted whole wheat grain flourless bread Tonus® (product of Trivit, Becej, Serbia) on DNA damage at various stages of progression through type 2 diabetes mellitus (T2DM). In addition to control (non-diabetic) subjects we analyzed obese, pre-diabetic and diabetic patients, for a total of four experimental groups. All subjects used to eat Mediterranean diet for at least two years before being included in our study. In each of four groups we had five persons practicing the Mediterranean diet but instead of bread they consumed flourless bread Tonus®. The DNA damage was evaluated on peripheral blood mononuclear cells by alkaline single cell gel electrophoresis (Comet) assay at the very beginning (before starting the Tonus® bread diet), and after exactly the three months of consumption of Tonus® bread. Statistical analysis revealed that only in patients with type 2 diabetes, Tonus® bread intake led to decreased DNA damage compared to the level of DNA damage of these patients before they started Tonus® bread diet. We assume that decrease of body weight and hyperinsulinemia caused by Tonus® bread in the diet might be one of the main causes of decreased DNA damage.

Apart from the efficiency of coumaphos against Varroa mites, its impact on the oxidative status and survival of the honey bee (Apis mellifera) was assessed. The research was conducted on hives from the same apiary, equalised regarding the number of bees, brood area and food storage. Based on Varroa infestation the hives were allotted to two groups: non-infested (N) and infested (I). Both groups were either treated (T) – NT and IT, or untreated (U) – NU and IU. The treatment of infested bees was controlled with a follow-up treatment with amitraz. The efficiency of coumaphos was 96-97%. This organophosphate had a negligible effect on bee survival, but it significantly affected their oxidative status: superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) activities, and the concentrations of malonyl dialdehyde (MDA). Coumaphos significantly (p˂0.0001) decreased SOD activity in non-infested bees, but increased it in those infested. By contrast, both CAT and GST activities, as well as MDA concentrations significantly increased (from p˂0.05 to p˂0.0001) after treatment in all groups, with the exception of IT, where it declined. Coumaphos in non-infested hives caused oxidative stress per se, not unlike varroa in infested colonies. However, in infested colonies it decreased oxidative stress, owing to its efficacy against Varroa mites and contributed to the recovery of bee colonies. In spite of its certain downsides, coumaphos remains an effective anti-varroa substance, but should be used with precaution, not to add to the effects of environmental factors which may cause red-ox misbalance.

Eight lovebirds of both sexes and different age were admitted showing alterations in behaviour and apathy. During the initial examination delamination of the beak was noted in all birds and discrete areas of alopecia in three of eight birds. Based on clinical experience, molecular diagnostic on PBFD, sequencing of obtained amplicons, and histopathological examination were performed. All birds tested positive on the presence of PBFD virus, despite the fact they did not exhibit expected clinical signs for PBFD. Sequencing results showed 100% match with sequences previously isolated from parrots with PBFD exhibiting classical PBFD manifestation. Histopathological examination showed similar findings as in previously described cases of PBFD in parrots. Other studies on psittacine birds correlate clinical manifestations and nucleotide variations with geographic localization. Our results indicate that the clinical manifestation of the disease is more dependent on bird species than on the genetic variation of the virus or the geographical distribution.

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.

Varroa destructor is the most important honey bee parasite. There are various methods used in the control of this mite, but none of them meets all requested criteria, to be safe, effective and easy to apply. The objective of this study was to evaluate the varroacidal efficacy of newly created plant-derived formulation Argus Ras (mixture of extracts of Sophora flavescens, Ginkgo biloba, Gleditsia chinensis and Teucrium chamaedrys) in a field trial. The investigation was conducted on 240 Apis mellifera colonies equalized in respect of brood amount, adult bee population and food reserves. Efficiency was evaluated by applying Argus Ras consecutively with two other acaricides, amitraz and oxalic acid. Average acaricidal efficacy of Argus Ras was 80.89%, being higher of other previously tested essential oils. Besides, it showed a potential in knocking down the mites resistant to other acaricides. It should not be neglected that Argus Ras requires a smaller number of treatments and financial investments than other formulations used for the control of Varroa mites.

Honey bees play a pivotal role in shaping ecosystems and sustaining human health as both pollinators and producers of health-promoting products. However, honey bee colony mortality is on the rise globally, driven by various factors, including parasites, pesticides, habitat loss, poor nutrition, and climate change. This has far-reaching consequences for the environment, economy, and human welfare. While efforts to address these issues are underway, the current progress in electron paramagnetic resonance (EPR) instrumentation affords using the immense potential of this magnetic resonance technique to study small samples such as honey bees. This paper presents the pioneering 2D in vivo EPR imaging experiment on a honey bee, revealing the ongoing redox-status of bees' intestines. This way, by monitoring the spatio-temporal changes of the redox-active spin-probes' EPR signal, it is possible to gain access to valuable information on the course of ongoing bees' pathologies and the prospect of following-up on the efficiency of applied therapies. Employing a selection of diverse spin-probes could further reveal pH levels and oxygen concentrations in bee tissues, allowing a noninvasive assessment of bee physiology. This approach offers promising strategies for safeguarding pollinators and understanding their biology, fostering their well-being and ecological harmony., (© 2024 Wiley Periodicals LLC.)

This review aims to present current knowledge on the effects of honey bee products on animals based on in vivo studies, focusing on their application in clinical veterinary practice. Honey's best-proven effectiveness is in treating wounds, including those infected with antibiotic-resistant microorganisms, as evidenced in horses, cats, dogs, mice, and rats. Propolis manifested a healing effect in numerous inflammatory and painful conditions in mice, rats, dogs, and pigs and also helped in oncological cases in mice and rats. Bee venom is best known for its effectiveness in treating neuropathy and arthritis, as shown in dogs, mice, and rats. Besides, bee venom improved reproductive performance, immune response, and general health in rabbits, chickens, and pigs. Pollen was effective in stimulating growth and improving intestinal microflora in chickens. Royal jelly might be used in the management of animal reproduction due to its efficiency in improving fertility, as shown in rats, rabbits, and mice. Drone larvae are primarily valued for their androgenic effects and stimulation of reproductive function, as evidenced in sheep, chickens, pigs, and rats. Further research is warranted to determine the dose and method of application of honey bee products in animals.

Thymol is a natural essential oil derived from the plant Thymus vulgaris L. It is known to be beneficial for human and animal health and has been used in beekeeping practice against Varroa mite for years. In this study, the genotoxic and antigenotoxic potential of thymol were evaluated on the honey bee ( Apis mellifera L.) continuous cell line AmE-711 for the first time. Using the Comet assay, three increasing concentrations (10, 100, and 1000 µg/mL) of thymol were tested. Negative control (non-treated cells) and positive control (cells treated with 100 µM H 2 O 2 ) were also included. The absence of thymol cytotoxicity was confirmed with the Trypan blue exclusion test. Thymol in the concentration of 10 µg/mL did not increase DNA damage in AmE-711 honey bee cells, while 100 and 1000 µg/mL concentrations showed genotoxic effects. For testing the antigenotoxic effect, all concentrations of thymol were mixed and incubated with H 2 O 2 . The antigenotoxic effect against was absent at all concentrations (10, 100, 1000 μg/mL) tested. Moreover, thymol enhanced the H 2 O 2 -induced DNA migration in the Comet assay. The obtained results indicate genotoxic effects of thymol on cultured honey bee cells suggesting its careful application in beekeeping practice to avoid possible negative effects on honey bees.

During their lifetime honey bees (Apis mellifera) rarely experience optimal conditions. Sometimes, a simultaneous action of multiple stressors, natural and chemical, results in even greater effect than of any stressor alone. Therefore, integrative investigations of different factors affecting honey bees have to be carried out. In this study, adult honey bees exposed to thiamethoxam in larval and/or adult stage and infected with Nosema ceranae were examined. Newly emerged bees from colonies, non-treated or treated with thiamethoxam, were organized in six groups and kept in cages. Thiamethoxam treated bees were further exposed to either thiamethoxam or Nosema (groups TT and TN), or simultaneously to both (group TTN). Newly emerged bees from non-treated colonies were exposed to Nosema (group CN). From both, treated and non-treated colonies two groups were organized and further fed only with sugar solution (groups C and TC). Here, we present the expression profile of 19 genes in adult worker honey bees comprising those involved in immune, detoxification, development and apoptosis response. Results showed that gene expression patterns changed with time and depended on the treatment. In group TC at the time of emergence the majority of tested genes were downregulated, among which nine were significantly altered. The same gene pattern was observed on day six, where the only significantly upregulated gene was defensin-1. On day nine most of analyzed genes in all experimental groups showed upregulation compared to control group, where upregulation of antimicrobial peptide genes abaecin, defensin-1 and defensin-2 was significant in groups TT and TTN. On day 15 we observed a similar pattern of expression in groups TC and TT exposed to thiamethoxam only, where most of the detoxification genes were downregulated. Additionally RNA loads of Nosema and honey bee viruses were recorded. We detected a synergistic interaction of thiamethoxam and Nosema, reflected in lowest honey bee survival., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This study is aimed at analysing biochemical and genetic endpoints of toxic effects after administration of adrenaline. For this purpose, the study was carried out on Wistar rats and three doses of adrenaline were used: 0.75 mg/kg, 1.5 mg/kg, and 3 mg/kg body weight. To achieve these aims, we investigated the effects of adrenaline on catalase (CAT), Cu, Zn-superoxide dismutase (SOD), malondialdehyde (MDA), nitrite (NO 2 -), carbonyl groups (PCC), and nitrotyrosine (3-NT). Total activity of lactate dehydrogenase (LDH), its relative distribution (LDH 1 -LDH 5 ) activity, level of acute phase proteins (APPs), and genotoxic effect were also evaluated. The obtained results revealed that all doses of adrenaline induced a significant rise in CAT activity, MDA level, PCC, NO 2 - , and 3-NT and a significant decrease in SOD activity compared to control. Adrenaline exerted an increase in total activity of LDH, LDH 1 , and LDH 2 isoenzymes. Further study showed that adrenaline significantly decreased serum albumin level and albumin-globulin ratio, while the level of APPs ( α 1 -acid glycoprotein and haptoglobulin) is increased. The micronucleus test revealed a genotoxic effect of adrenaline at higher concentrations (1.5 mg/kg and 3 mg/kg body weight) compared to untreated rats. It can be concluded that adrenaline exerts oxidative and nitrative stress in rats, increased damage to lipids and proteins, and damage of cardiomyocytes and cytogenetic damage. Obtained results may contribute to better understanding of the toxicity of adrenaline with aims to preventing its harmful effects.