Rajmund Sokół

Detection of Nosema spp. in Worker Bees of Different Ages During the Flow Season

Abstract The aim of this study was to identify which Nosema species infect those Apis mellifera worker bees performing different functions in the colony. Samples were taken from different places inside and outside the hive, in the honey flow season. In February 2010, winter hive debris from 30 colonies was analyzed, and based on the microsporidian species identified by multiplex PCR. The following bee colonies (none of which displayed clinical symptoms of the disease) were selected for further analyses to determine the occurrence of microsporidian parasites: 1) colony A/C infected with Nosema apis and N. ceranae (mixed infection), 2) colony A infected with N. apis, 3) colony C - infected with N. ceranae, and 4) colony K - the control, which was free of infection. Between April and August, 20 nurse bees from frames of open brood, and 20 forager bees returning to the hive from pollen-collecting trips were randomly selected from each colony at 30-day intervals. The results of the study indicate that the microsporidian species is determined not only by the type of worker bee (sampling site), but also by the period (month) of the sample collection. Our findings also suggest that regardless of the type of initial infection, bees infected by different microsporidian species and bees free from infection can coexist in colonies. Streszczenie Celem badań było ustalenie, który gatunek Nosema występuje u robotnic Apis mellifera pobieranych z różnych miejsc ula w sezonie pasiecznym. W lutym 2010 roku zbadano osypy zimowe pszczół z 30 rodzin pszczelich. W zależności od stwierdzonego gatunku sporowca (metodą multiplex PCR) dalszą ocenę występowania gatunku sporowców prowadzono w następujących grupach (rodzinach pszczelich bez objawów klinicznych choroby): AC - zainfekowana Nosema apis i Nosema ceranae (infekcja mieszana), A - zainfekowana N. apis, C - zainfekowana N. ceranae oraz grupa kontrolna K, wolna od infekcji. Z każdej rodziny od kwietnia do sierpnia w odstępach 30 dniowych pobierano do badań losowo po 20 robotnic z dwóch miejsc: robotnice ulowe, bezpośrednio z plastrów z czerwiem otwartym oraz robotnice lotne - robotnice powracające do uli z pyłkiem. Wykazaliśmy, że nie tylko rodzaj robotnic (miejsce pobrania), ale i okres (miesiąc) pobrania prób do badań ma wpływ na rodzaj wykrywanego gatunku sporowca, a także, że niezależnie od rodzaju infekcji początkowej (w osypie zimowym) w rodzinach pszczelich mogą współistnieć osobniki zainfekowane różnymi gatunkami sporowca oraz wolne od infekcji.

Estimation of infection of internal parasites in horses from different type of farms.

Studies were carried out in year 2014 during the pasture period (from April to October) in Warmia and Mazury Region. Fecal samples were taken from cold- and warmblood horses from individual and agrotouristic farms with the different housing, feeding and pasture- care practices. Total of 512 horses were examined (320 mares, 170 geldings and 22 stallions). In the group of 185 horses from individual farms, 119 animals (64.3%) were infected with gastro-intestinal parasites. Among the 372 horses from agrotouristic farms 169 (51.7%) were infected with parasites. Most of the animals expelled the eggs of Cyathostominae. In some individuals occurred eggs of Strongylus spp., Parascaris equorum, Strongyloides westeri and tapeworm of Anoplocephala. The number of infected horses from agrotouristic farms was lower than from individual farms, probably due to more regular deworming (usually 2 times a year) and bigger care paid to cleaning pastures.

Biochemical status of feral honey bees (Apis mellifera) infested with various pathogens

Feral bees colonizing large forest habitats without human intervention are also affected by diseases. Infected bees that survive several seasons constitute an interesting object of scientific inquiry. Little is known about the health and status of feral honey bee populations. The pathogens infesting/infecting bee colonies and the biochemical status of unhealthy bees need to be studied to prevent the global decline in bee populations. Honey bees inhabiting hollow trees in forest reserves of north-eastern Poland, infected with Nosema ceranae or infested with Varroa destructor and/or infected with the deformed wing virus (DWV), were analyzed. Differences in the analyzed biochemical parameters were noted in groups infected/infested with various pathogens relative to the control group. The most significant variations were observed in antioxidant parameters: lower total antioxidant status (TAS) (excluding group N. ceranae), lower activity of superoxide dismutase (SOD) and peroxidase (POX) (excluding group N. ceranae + DWV), higher activity of glutathione transferase (GST), and higher glutathione (GSH) levels in groups infected with N. ceranae. Glycogen concentration was higher in DWV-infected bees. In comparison with bees infested with V. destructor only, co-infested groups were characterized by fluctuations in carbohydrate concentration, subject to the co-infesting pathogen. The presence of a single pathogen has a less detrimental effect on most metabolic parameters in feral honey bees than concomitant infections with several pathogens. The results of this study suggest that co-infestations with V. destructor and DWV in feral honey bees have more serious consequences than co-infections with Nosema sp. and DWV.

Invasive forms of canine endoparasites as a potential threat to public health – A review and own studies

[b]Abstract [/b] Dogs serve as the vectors of serious zoonotic parasitic diseases. In the month of May 2012 - 2014, 339 dog faeces samples from seven public sites in Chełmno, a town in northern Poland, were collected and examined to determine the gastrointestinal parasite fauna of dogs. Each faecal sample was dissected with a needle, checked for tapeworm segments and examined for parasite eggs and oocysts using the flotation and decantation method and a modified Baermann technique. Differences were observed in the degree of parasite species occurrence. The most dominant were [i]Toxocara canis[/i] and Ancylostomatidae. The detected species included: [i]T. canis [/i]and [i]Toxascaris leonina[/i] eggs (23.4% and 10.2%, respectively), as well as eggs from the[i] Ancylostomatidae[/i] family (16.2%),[i] Trichuris vulpis [/i]eggs (6.6%), [i]Taenia[/i] type eggs (4.6%),[i] Dipylidium caninum[/i] (5.2%) and [i]Cystoisospora [/i](Isospora) spp. oocysts (10.9%).

Evaluation of the Effectiveness of Selected Treatments of Nosema Spp. Infection by the Hemocytometric Method and Duplex Pcr

Abstract Recent years have witnessed an increase in the mortality of honey bees in many regions of the world. The observed decrease in the bee population results from a combination of factors, and microsporidian parasites Nosema apis and N. ceranae are among the main contributors. Those parasites cause a microsporidian infection that shortens the lifespan of bees and reduces the productivity of bee colonies. The aim of this study was to evaluate the effectiveness of Nozevit, Api Herb and ApiX (acetylsalicylic acid + Artemisia absinthium L. extract) in the control of infections caused by Nosema spp. in a field experiment. Two groups of worker bees were evaluated - hive bees and forager bees returning to the hive. The effect of the analyzed therapies on the number of spores and the microsporidia species were analyzed by the hemocytometric method and duplex PCR. A statistical analysis revealed that the applied treatments had reduced the number of spores by 31.15% on average. In hive bees, Nosema spp. infection was most effectively reduced by Nozevit (67.85%) and ApiX (63.36%). Coinfections (N. ceranae and N. apis) were affirmed in all bee samples before treatments. However, after the treatments, single infection of N. apis and N. ceranae were detected. The tested treatments were more effective in the control of N. apis than N. ceranae.

The resistance of Eimeria spp. to toltrazuril in black grouse (Lyrurus tetrix) kept in an aviary

ABSTRACT The black grouse is a critically endangered species in Europe. Aviary breeding and re‐introduction programs have contributed to the reinstatement of black grouse populations. Parasitic diseases can influence the species’ behavior, choice of habitats, and survival rates. Protozoa of the genus Eimeria are the most prevalent parasites in black grouse. The aim of this study was to evaluate the effectiveness of toltrazuril (TOL) against Eimeria spp. in black grouse kept in an aviary. Eimeria spp. oocysts (mean OPG for all birds = 7,047.84; SD = 5,625.05) were detected in feces samples in a routine parasitological examination. Parasitic infections were treated with toltrazuril Baycox 2.5% (Bayer, Leverkusen, Germany) at 1 ml/1 l H2O administered per os with drinking water twice a day for 2 d, 12 h/24 h. High oocysts counts persisted upon the second parasitic examination. Therefore, the birds were divided into 3 groups in 3 existing aviaries. Toltrazuril was administered according to the same protocol, and its uptake with drinking water was monitored. A parasitological examination conducted 5 d later revealed the presence of parasites, and the TOL dose was increased to 3 ml/1 l H2O. When oocysts were detected in a successive exam, the TOL dose was further increased to 5 ml/1 l H2O. The increase in TOL dose did not eliminate parasites, therefore, the treatment was discontinued. The fecal egg count reduction test (FECRT) revealed that 1 ml of TOL reduced fecal oocysts counts (OPG) by 0.02%, the second application of the same TOL dose reduced OPG by 0%, 3 ml of TOL reduced OPG by 0%, and 5 ml of TOL reduced OPG by 65.7%. The applied doses of TOL did not induce significant differences in the fecal oocysts counts of any of the identified species of coccidia. The results indicate that the coccidia species infecting black grouse have natural resistance to TOL.

Estimation of the influence of selected products on co-infection with N. apis/N. ceranae in Apis mellifera using real-time PCR

Abstract To protect the world’s honey bee population many scientific centres are searching for products and methods that control nosemosis. Real-time PCR was used to assess infection level in worker bees infected with Nosema spp. in bee colonies co-infected with Nosema apis and Nosema ceranae after the administration of three products (Nozevit, ApiHerb and ApiX) and sugar syrup. The study was conducted in the field condition therefore there was no possibility to affect the number of spores in the selected material. The study demonstrated considerable differences in the number of spores of individual Nosema spp. in the analysed samples of bees. HSD Tukey’s test showed that the statistically significant effect on limiting the N. apis invasion had ApiX (p – 0.049). Nozevit, Apiherb and syrup showed no statistically significant effect on reducing the amount of N. apis spores. The same test showed that the statistically significant effect on limiting the N. ceranae invasion had: Nozevit (p – 0.014), Apiherb (p – 0.032), ApiX (p – 0.034) and syrup (p – 0.033). There was no statistically significant decrease in the N. ceranae spores in the control group.