Exposure of honey bee larvae to thiamethoxam and its interaction with Nosema ceranae infection in adult 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

Efficacy of plant-derived formulation argus ras in varroa destructor control

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

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

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

Potential of Fumagillin and Agaricus blazei Mushroom Extract to Reduce Nosema ceranae in Honey Bees

Depending on the infection level and colony strength, Nosema ceranae, a microsporidian endoparasite of the honey bee may have significant consequences on the health, reproduction and productivity of bee colonies. Despite exerting some side effects, fumagillin is most often used for Nosema control. In this study, in a cage experiment, N. ceranae infected bees were treated with fumagillin or the extract of Agaricus blazei mushroom, a possible alternative for Nosema control. Bee survival, Nosema spore loads, the expression levels of immune-related genes and parameters of oxidative stress were observed. Fumagillin treatment showed a negative effect on monitored parameters when applied preventively to non-infected bees, while a noticeable anti-Nosema effect and protection from Nosema-induced immunosuppression and oxidative stress were proven in Nosema-infected bees. However, a protective effect of the natural A. blazei extract was detected, without any side effects but with immunostimulatory activity in the preventive application. The results of this research suggest the potential of A. blazei extract for Nosema control, which needs to be further investigated

The Effects of Antioxidants Provided with Feed on Certain Quality Parameters of Bull Semen under Heat Stress Conditions

The aim of the current research was to assess the effects of the feed additive made of lyophilised melon juice (source of superoxide dismutase, SOD) and inactivated live Saccharomyces cerevisiae (strain R397) cells added to the feed via the product containing high levels of organically bound selenium (source of selenium-dependant glutathione peroxidase, Se-GPx) on the semen quality of bulls in heat stress conditions. The 15 bulls chosen for the experiment were assigned to three equal groups (control-group C; treated group M, given the source of SOD; and group A, treated with the source of Se-GPx). The research was conducted in summer. The activities of SOD and Se-GPx in seminal plasma were determined spectrophotometrically. Computer-assisted semen analysis was done to determine the sperm counts, motility and velocity. The temperature and humidity were recorded with a digital data logger. The average SOD activity in the control bulls was significantly lower than in M (p0.05). In all of the three groups with the increase in THI there was an increase in GPx activity (p>0.05). It can be concluded that in all of the three groups of bulls there was an increase in the activity of both enzymes in the seminal plasma, but the increase was significantly lower in the control. Thus, the antioxidative capacity of the seminal plasma of untreated bulls was proven to be lower in comparison with those of the treated animals

Genome-wide analysis of milk production traits and selection signatures in Serbian Holstein-Friesian Cattle

To improve the genomic evaluation of milk-related traits in Holstein-Friesian (HF) cattle it is essential to identify the associated candidate genes. Novel SNP-based analyses, such as the genetic mapping of inherited diseases, GWAS, and genomic selection, have led to a new era of research. The aim of this study was to analyze the association of each individual SNP in Serbian HF cattle with milk production traits and inbreeding levels. The SNP 60 K chip Axiom Bovine BovMDv3 was deployed for the genotyping of 334 HF cows. The obtained genomic results, together with the collected phenotypic data, were used for a GWAS. Moreover, the identification of ROH segments was performed and served for inbreeding coefficient evaluation and ROH island detection. Using a GWAS, a polymorphism, rs110619097 (located in the intron of the CTNNA3 gene), was detected to be significantly (p < 0.01) associated with the milk protein concentration in the first lactation (adjusted to 305 days). The average genomic inbreeding value (FROH) was 0.079. ROH islands were discovered in proximity to genes associated with milk production traits and genomic regions under selection pressure for other economically important traits of dairy cattle. The findings of this pilot study provide useful information for a better understanding of the genetic architecture of milk production traits in Serbian HF dairy cows and can be used to improve lactation performances in Serbian HF cattle breeding programs

Genotoxic Potential of Thymol on Honey Bee DNA in the Comet Assay

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 H2O2) 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 H2O2. The antigenotoxic effect against was absent at all concentrations (10, 100, 1000 μg/mL) tested. Moreover, thymol enhanced the H2O2-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. © 2023 by the authors

Diet supplementation helps honey bee colonies in combat infections by enhancing their hygienic behaviour

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.The supplement “BEEWELL AminoPlus” induced a significant and consistent increase of the hygienic behavior in spite of the negative effects of N. ceranae and viral infections. N. ceranae and viruses significantly and consistently decreased hygienic behavior, but also threatened the survival of bee colonies. The tested supplement showed anti-Nosema effect, since the N. ceranae infection level significantly and consistently declined onl

Da li je simptomatologija PBFD više zavisna od vrste domaćina nego od soja virusa? – Slučaj 8 rozenkolisa

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.Osam rozenkolisa oba pola različite starosti su primljeni na pregled zbog uočenih promena u ponašanju i apatije. Tokom inicijalnog pregleda kod svih ptica je uočena delaminacija kljuna, a kod tri jedinke bila su prisutna i diskretna polja alopecije. Na osnovu kliničkog iskustva postavljena je sumnja na oboljenje bolest kljuna i perja te je urađena molekularna dijagnostika na pristutvo virusa bolesti kljuna i perja, sekvencioniranje dobijenih amplifikata i patohistološko ispitivanje. Rezultati ispitivanja su potvrdili prisustvo virusa u uzorcima od svih ispitivanih jedinki, poklapanje nukleotidnih sekvecni od 100% sa uzorcima od drugih ptica sa drugačijom kliničkom slikom, a patohistološke promene su identične onima opisanim kod bolesti kljuna i perja. Studije brojnih autora povezivale su kliničku manifestaciju bolesti sa nukleotidnim varijacijama ili geografskom lokalizacijom. Međutim, opisani rezultati bi mogli da ukazuju da je kliničko ispoljavanje bolesti više zavisno od vrste domaćina nego od genotipa virusa koji bolest izaziva i od teritorije na kojoj se bolest ispoljava