Gastrointestinal Parasites in Owned Dogs in Serbia: Prevalence and Risk Factors.

Dogs are the most popular pets worldwide. Close contact between dogs and people increases the risk of transmission of various zoonotic parasitic infections. Given the importance of veterinary medicine in preserving the One Health concept, the aim of this research was to identify intestinal parasites that may have zoonotic potential and to evaluate risk factors (individual and environmental). The research was conducted in Serbia in 2022 and 2023 on 382 owned dogs, using qualitative methods of coprological examination with a concentration on parasitic elements. The overall prevalence of intestinal parasites was 62.6%, with the following detected: protozoa: Cystoisospora spp. (9.2%), Sarcocystis spp. (4.5%), Neospora caninum/Hammondia spp. (3.7%), Giardia intestinalis (11.8%); nematoda: Toxocara canis (11.5%), Toxascaris leonina (4.2%), family Ancylostomatidae (38.0%), Trichuris vulpis (21.5%), Capillaria spp. (10.5%); trematoda: Alaria alata (1.6%) and cestodes from the Taeniidae family (1.3%). Factors like age, size and coat length, as well as the way of living, attitude and diet were linked to a significantly higher (p < 0.05) prevalence of intestinal parasites. Based on the results of coprological diagnostics, this research indicates the importance of educating dog owners, conducting routine parasitological tests on their pets and regular deworming strategies.

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).

The presence of malignant ovine babesiosis in Bosnia and Herzegovina indicates a possible emerging risk for Balkan region.

Malignant ovine babesiosis or ovine babesiosis caused by Babesia ovis is an important hemoprotozoan tick-borne disease of sheep. After 70 years, a study was conducted in Bosnia and Herzegovina, in the geographical areas of Podrinje and Eastern Herzegovina, aiming to molecularly identify and determine the presence of B. ovis infection in the sheep population. From 2019-2021, a clinical and molecular study was conducted in 53 flocks, and 192 sheep were examined, divided into two groups: clinically suspected and asymptomatic sheep. The presence of B. ovis was confirmed by PCR in blood samples of 75 % and 11.21 % of clinically affected and asymptomatic sheep, respectively. The majority of clinical cases of malignant ovine babesiosis were confirmed in the Rudo epidemiological unit (78.78 %) within the Podrinje region indicating typical seasonal pattern of disease occurrence and endemic focus. Rhipicephalus bursa was only tick species identified in Podrinje and Eastern Herzegovina. Acute form of disease was observed with dominant clinical signs of depression, fever, loss of appetite and respiratory distress. Fatal outcome was recorded in 12.28 % of cases. This study describes the molecular detection of B. ovis in sheep in Bosnia and Herzegovina for the first time. To the authors knowledge, this survey represents a report of the highest number of clinical cases of malignant ovine babesiosis in Europe. Since the disease has been recorded in the south-eastern border of Bosnia and Herzegovina, there is a high risk of disease transmission to a wider area of the Balkan region.

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.

Plant-based supplement containing B-complex vitamins can improve bee health and increase colony performance.

It is common knowledge that nutritive stress resulting from decreased diversity and quality of food, pollution of food sources and beekeeping errors may lead to increased susceptibility of bees to pathogens and pesticides. The dearth of adequate food is frequently compensated with supplements. Thus, this research was aimed to study the effects of the plant-based supplement B + on colony strength (assessed according to open and sealed brood area, honey and pollen/bee bread reserves, and the number of adult bees). In addition, Nosema ceranae spores and viruses were quantified and the level of infestation with Varroa destructor assessed. The experiment was conducted in late summer and early spring. In colonies which were given B + in feed a significant increase (p < 0.05) in the parameters of colony strength were noticed in comparison to the control (colonies fed on sugar syrup). Moreover, it was proven that the bees from these colonies had significantly lower (p < 0.05) N. ceranae spore counts, and acute bee paralysis, deformed wing and sacbrood virus loads. Our results suggest that the addition of B + supplement to the colonies provide them with nutrients, contribute to their strengthening, might prevent nutritive stress and increase the success of bees in combating pathogens.

Quantitative PCR assessment of Lotmaria passim in Apis mellifera colonies co-infected naturally with Nosema ceranae.

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.