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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.

Mushroom extracts have been shown to contain bioactive compounds that confer health benefits to humans and animals, including honey bees. Honey bees are often challenged simultaneously with several abiotic (e.g., pesticides and poor nutrition) and biotic (e.g., parasites and pathogens) stressors. Mushroom extracts, for example, those made from Agaricus bisporus, may hold the potential to mitigate the negative effects of these stressors through actions on the honey bee's immune system, metabolism, and other physiological processes. Exploring the health benefits of mushroom extracts for honey bees requires a basic understanding of their bioactive properties at the cellular level. The aim of the present study was to evaluate, for the first time, the genotoxic and antigenotoxic potential of A. bisporus on the honey bee cell line AmE-711 using the comet assay. Three increasing concentrations of A. bisporus water extract (100, 200, and 400 µg/mL) were tested. The cells in the negative control group were not treated either with A. bisporus extract or with H2O2. In the positive control group, DNA damage was induced with 100 µM H2O2. For antigenotoxic effect, tested extracts were mixed and incubated with H2O2. None of the tested concentrations exerted genotoxic potential, but all showed antigenotoxic effects against H2O2-induced DNA damage. The concentration of 200 μg/mL A. bisporus extract was the most effective in its action against DNA damage. To conclude, A. bisporus extract did not demonstrate genotoxic effects but showed promising antigenotoxic properties. AmE-711 cell line may serve as a cell culture system for genotoxicity investigations. [ABSTRACT FROM AUTHOR]

Simple Summary: Microsporidia Nosema ceranae is an obligate intracellular parasite of the honey bee and causes nosemosis, a disease with negative effects on the health, reproductive and productive capabilities of bee colonies. The aim of this study was to investigate the effects of a plant-based supplement branded as "B+" on honeybees in a laboratory experiment. Food supplement branded as "B+" showed the potential to positively influence bee survival. Supplemental feeding reduced N. ceranae infection level. Treatment significantly decreased oxidative stress in Nosema-infected bees. Vitellogenin gene expression was the highest in the supplement-fed group. 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). [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the length of the contraceptive period after the application of deslorelin implant (4.7 mg) in juvenile, prepubertal bitches and assess the effects on the hormonal status (P4, E2 and melatonin), as well as on the occurrence of normal physiological estrus. Twelve crossbred prepubertal female dogs (4 months of age) were divided into the two groups, the treated group (DESLO) and control (placebo) group (C). Hormone assays, vaginal microbiological and cytological smears and the measurements of electrical resistance of cervical mucus were performed once in 3 weeks until the first signs of proestrus, and every day during estrus. GnRH implant significantly postponed time of onset of estrus (633 ± 30,38 days vs. 143.80 ± 52,30 days, P

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 5mM 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 sucrosewater 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 5mM 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. [ABSTRACT FROM AUTHOR]

The objective was to expand and update the knowledge on the presence and genotype diversity of porcine circoviruses 2 and 3 (PCV2 and PCV3) in the wild boar populations from the hunting grounds in northeastern Serbia. The presence of PCV3 was not determined, and PCV2 was confirmed in 40.32% of the organ samples from 124 wild boars hunted from 2018 to 2019, indicating their significance in virus circulation since traditional pig farms with irregular PCV2 vaccination strategies are widespread in this region. The most prevalent genotype was PCV2d, followed by PCV2b and PCV2a in 55.6%, 38.9%, and 5.5% of the examined samples, respectively. Nucleotide sequences of the detected strains were homogenous within the genotype and clustered within the subgroups PCV2d-2, PCV2b-1A/B, and PCV2a-2D with high identity to European, Chinese, and Serbian domestic pig sequences suggesting their origin. Wild boars presented with no clinical or pathological signs of infection, implying that these animals might be less susceptible to disease, particularly since the cofactors present in pig farming systems that support the disease development are absent in the wild. The high PCV2 detection frequency demonstrates the importance of wildlife monitoring to track virus population dynamics, especially in regions with free-range pig farming in order to plan adequate disease control strategies. [ABSTRACT FROM AUTHOR]

The aim of this study was to examine the seroprevalence of Leptospira spp. in dogs and red foxes in the entity of the Republic of Srpska, Bosnia and Herzegovina, after heavy rainfall and floods in 2014 and for the two years thereafter. The seroepidemiological study involved testing serum samples from dogs (n = 98) and foxes (n = 112) using MAT (microscopic agglutination test).

The present work aimed to study the goat farms concerning different hygiene-sanitary and health management practice. Also, bulk tank milk of Alpine breed goats was used to investigate changes in the composition of milk during lactation on six farms in Vojvodina Region. The results showed that although 96.67% of farmers were familiar with mastitis, approximately 56.67% of farmers reported a lack of knowledge about subclinical mastitis and therefore not intensely monitoring udder health during the lactation period. Somatic cell count (SCC) and total bacteria count (TBC) were the most variable traits in our research and lactation average SCC between investigated farms showed a significant difference (P < 0.01) (a range from 1088×10?/ml to 3129×10?/ml). At the same time, TBC was found in a wider range, from 66×10?/ml to 3508×10?/ml. Furthermore, SCC and TBC often did not meet the recommended quality level. Therefore, during the entire lactation period, 63% of the milk samples contained above 1.5 million SCC/ml and 37% of the samples were contained TBC above 1.5 million CFU/ml. Results also confirmed considerable seasonal variations of goat milk composition and the milk fat was the component that most varied (cv: 33.16%). It was observed that organic in comparison to conventional milk chemical composition, was significantly different, especially in terms of protein content (P < 0.0001). In conclusion, goat dairy production could be of considerable importance in Vojvodina but dairy goat sector requires a more systemic approach, better hygiene milking conditions and implemented prevention and control programmes during the milking routine. [ABSTRACT FROM AUTHOR]

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.

A recently described trypanosomatid species Lotmaria passim and the microsporidium Nosema ceranae infect the honey bee ( Apis mellifera ), but the interspecific dynamic of these two common gut parasites is unknown. In this study, a real-time qPCR assay was developed to enable the specific detection and quantification of L. passim . The annual dynamics of N. ceranae and L. passim infections were evaluated in ten A. mellifera colonies naturally infected with both parasites at one apiary in Serbia from March 2016 to March 2017. Ten samples (60 bees abdomens) were taken from each colony on 8 sampling occasions. L. passim infection level was evaluated with qPCR, while N. ceranae infection was measured by spore counts. N. ceranae infection level was significantly higher in comparison with that of L. passim (spore or cell equivalents/bee, respectively). Significant positive correlation between infection levels of the parasite species indicates their similar annual dynamics, whilst the differences in the levels of infection between particular months point to a seasonal pattern in the incidence of both parasites. The assay which has been developed and validated creates opportunity for detailed study of L. passim infection kinetics and the improvement in the management practices in beekeeping related to these two parasites. [ABSTRACT FROM AUTHOR]

Introduction. Glutathione peroxidases are selenoenzymes which have a crucial role in the protection of animals against oxidative stress. Materials and Methods. From September 2017 to April 2018, a group of eight red-eared sliders were admitted at the Clinic for Small Animals, Faculty of Veterinary Medicine, University of Belgrade for elective diagnostic celioscopy. The turtles were of unknown age, weight from 1.20 kg to 1.86 kg. The anesthesia protocol involved using ketamine and medetomidine, both at a low dosage (10 mg kg-1 and 0.1 mg kg-1, respectively) as induction, after which anesthesia was maintained using isoflurane at 3% (vapor setting) in 100% oxygen (0.5 L min-1). Medetomidine was reversed with atipamezole (0.2 mg kg-1), given intramuscularly. The elective celioscopy was performed according to standard protocols. One day prior to anesthesia, heparinized blood samples were taken using the subcarapacial venous plexus for venipuncture. The second sampling took place three hours after the anesthetics were administered. Results and Conclusions. GPx3 activity in the blood plasmas (n=8) was measured by the coupled test as described by Günzler et al. (1974). Data were tested for normality by the Shapiro-Wilk normality test and the groups were compared using a paired t-test. Blood plasma GPx3 activity was significantly higher (p=0.009) after a three-hour recovery period from inhalation anesthesia performed for elective diagnostic celioscopy, than before anesthesia. The measured post-anesthesia GPx3 activities were, on average, 80% higher than the measurements prior to anesthesia. It can be concluded that the statistically significant increase in the activity of plasma GPx3 from 91.02±36.05 mKat L-1 prior to anesthesia to 160.21±58.94 mKat L-1 three hours after anesthesia is due to the change in oxygen saturation. This is increased to 100% during the procedure, thus exposing the turtles to conditions of high oxygen saturation.

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.

Introduction. Trichinellosis is a disease in humans caused by parasites of the genus Trichinella, and these roundworms can occur in a variety of animals (over one hundred mammal species). Members of the genus Trichinella are present in almost all continents and in all climate zones. Intensive studies on the eradication of this disease have been going on for a long period, but despite the finances invested in research projects, trichinellosis is still present in the 21st century and poses a major health issue all over the world. According to current scientific estimations, there are over 27 million Trichinellainfected people in the world. The aim of our study was to determine the distribution and trends for Trichinella infection in pigs and trichinellosis in humans in Serbia between 1994 and 2018. Materials and Methods. Data for the 25-year surveillance period of Trichinella cases registered in pigs and humans in Serbia was gathered from the Veterinary Directorate and from the Institute of Public Health of the Republic of Serbia. The data obtained was analysed with the relative numbers of structure and dynamics, indices and descriptive statistical indicators. Results and Conclusions. During the research period, 14,837 pigs were diagnosed as infected with Trichinella. Out of this number, 87.31% of pigs were identified in the five epizootiological regions, and only 12.69% were diagnosed in the non-epizootiological regions in Serbia. During the period 1994-2018 in Serbia, a total of 6,850 people were treated for Trichinella infection. Out of this number, 4,153 (60.63%) people were from the five epizootiological regions. The trend-line describing the presence of Trichinella in pigs was defined by a fourth degree polynomial function. Meanwhile, the trend-line describing the presence of trichinellosis in humans was defined by a sixth degree polynomial function. Trichinellosis in Serbia is most common during the winter season, from December to March. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR 31034: Selected biological hazards to the safety/quality of food of animal origin and the control measures from farm to consumer]

Introduction. Trichinellosis is a cosmopolitan anthropozoonosis, and is a group of systemic diseases caused by larval forms of Trichinella. This is one of the oldest and most controversial parasitic zoonoses that has been in the forefront of veterinary, medical and biological research for many years. Parasites from the genus Trichinella were diagnosed in more than one hundred species of domestic and wild mammals and birds. The aim of this study was economic evaluation of a program for monitoring and controlling Trichinella in pigs. Materials and Methods. A systematic pest rodent control program deemed suitable for monitoring and controlling Trichinella in pigs was implemented in a representative test zone. The results of the pest rodent control program were analyzed using cost-benefit analysis to determine if a similar program would be suitable for a larger land area. Results and Conclusions. Prior to pest control, the incidence of Trichinella in pigs living in the test zone increased annually, while after systematic pest rodent control, the incidence of Trichinella-positive pigs decreased. These results, as well as relevant economic indicators, enabled us to design two mathematical models describing the control and monitoring of Trichinella in a larger land area. In this area, Srem Region, Serbia, a suitable Trichinella control program in pigs is clearly justified because the profit to cost ratio was positive for both models. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR 31034: Selected biological hazards to the safety/quality of food of animal origin and the control measures from farm to consumer]

The study was to conduct to evaluate the chemical composition and nutritive values of feedstuffs (forages and concentrate mixtures) used for dairy goats nutrition in Vojvodina. Samples were collected from six farms, including one organic farm. The results showed that the relative feed values of analyzed forages were in the range of good, medium to lower quality. Average protein content from lowest to highest for investigated forages was: corn silage (Zea Mays) (65.37-82.57g kgˉ¹DM), alfalfa haylage (Medicago sativa L.) (159.99-184.17g kgˉ¹DM), pasture (185.30g kgˉ¹ DM), and alfalfa hay (Medicago sativa L.) (167.48-203.60g kgˉ¹DM). The non-fibre carbohydrates and protein content most varied in organic hay samples (cv: 29.25% and 19.09%, respectively). Generally, feedstuffs used in organic nutrition, including organic concentrate, were of lower nutritional quality and moreover contained higher amounts of crude fibre and lignin. Especially, a high source of variation was observed in investigated concentrate mixtures for the crude protein content (p

Simple Summary: Nosema ceranae affects honey bee (Apis mellifera L.) causing nosemosis disease that often induces serious problems in apiculture. Antibiotic fumagillin is the only licenced treatment against nosemosis, but its effectiveness is questioned and its usage associated with risk of bee mortality and appearance of residues in bee products. In search for alternative treatment for the control of nosemosis, water crude extract of Agaricus bisporus was tested on bees in laboratory (cage) experiments. Bee survival and food consumption were monitored together with Nosema infection level and expression of five genes (abaecin, hymenoptaecin, defensin, apidaecin, and vitellogenin) were evaluated in bees sampled on days 7 and 15. Apart from the gene for defensin, the expression of all tested genes was up-regulated in bees supplemented with A. bisporus extract. Both anti-Nosema and immune protective effects of A. bisporus extract were observed when supplementation started at the moment of N. ceranae infection or preventively (before or simultaneously with the Nosema infection). Agaricus bisporus water crude extract was tested on honey bees for the first time. The first part of the cage experiment was set for selecting one concentration of the A. bisporus extract. Concentration of 200 µg/g was further tested in the second part of the experiment where bee survival and food consumption were monitored together with Nosema infection level and expression of five genes (abaecin, hymenoptaecin, defensin, apidaecin, and vitellogenin) that were evaluated in bees sampled on days 7 and 15. Survival rate of Nosema-infected bees was significantly greater in groups fed with A. bisporus-enriched syrup compared to those fed with a pure sucrose syrup. Besides, the anti-Nosema effect of A. bisporus extract was greatest when applied from the third day which coincides with the time of infection with N. ceranae. Daily food consumption did not differ between the groups indicating good acceptability and palatability of the extract. A. bisporus extract showed a stimulative effect on four out of five monitored genes. Both anti-Nosema and nutrigenomic effects of A. bisporus extract were observed when supplementation started at the moment of N. ceranae infection or preventively (before or simultaneously with the infection). [ABSTRACT FROM AUTHOR]

Commercially and traditionally managed bees were compared for oxidative stress (superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST) and malondialdehyde (MDA)), the prevalence of parasites (Lotmaria passim, Crithidia mellificae and Nosema ceranae/apis) and social immunity (glucose oxidase gene expression). The research was conducted on Pester plateau (Serbia—the Balkan Peninsula), on seemingly healthy colonies. Significant differences in CAT, GST and SOD activities (p < 0.01), and MDA concentrations (p < 0.002) were detected between commercial and traditional colonies. In the former, the prevalence of both L. passim and N. ceranae was significantly (p < 0.05 and p < 0.01, respectively) higher. For the first time, L. passim was detected in honey bee brood. In commercial colonies, the prevalence of L. passim was significantly (p < 0.01) lower in brood than in adult bees, whilst in traditionally kept colonies the prevalence in adult bees and brood did not differ significantly. In commercially kept colonies, the GOX gene expression level was significantly (p < 0.01) higher, which probably results from their increased need to strengthen their social immunity. Commercially kept colonies were under higher oxidative stress, had higher parasite burdens and higher GOX gene transcript levels. It may be assumed that anthropogenic influence contributed to these differences, but further investigations are necessary to confirm that. [ABSTRACT FROM AUTHOR]

In this study, honey bees collected in Serbia over 9 consecutive years (2007–2015) were retrospectively surveyed to determine the prevalence of eukaryotic gut parasites by molecular screening of archival DNA samples. We developed species-specific primers for PCR to detect the two known honey bee trypanosomatid species, Crithidia mellificae and the recently described Lotmaria passim . These primers were validated for target specificity under single and mixed-species conditions as well as against the bumblebee trypanosomatid Crithidia bombi . Infections by Nosema apis and Nosema ceranae (Microsporidia) were also determined using PCR. Samples from 162 colonies (18 from each year) originating from 57 different localities were surveyed. L . passim was detected in every year with an overall frequency of 62.3% and annual frequencies ranging from 38.9% to 83.3%. This provides the earliest confirmed record to date for L . passim and the first report of this species in Serbia. N . ceranae was ubiquitous, occurring in every year and at 95.7% overall frequency, ranging annually from 83.3% to 100%. The majority of colonies (60.5%) were co-infected with L . passim and N . ceranae , but colony infections by each species were statistically independent of one another over the nine years. Although C . mellificae and N . apis have both been reported recently at low frequency in Europe, neither of these species was detected in Serbia. These results support the hypothesis that L . passim has predominated over C . mellificae in A . mellifera during the past decade. [ABSTRACT FROM AUTHOR]

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).