A Preliminary Study on "Personalised Treatment" against Varroa destructor Infestations in Honey Bee (Apis mellifera) Colonies.

The requirement for the protection of bee colonies against Varroa destructor invasions has been noted by many breeders and is included as an aspect of the development of beekeeping. This research aimed to check the effect of the development of a colony exposed to laying eggs (brood surface) by queen bees with similar chemical potential (sisters) on the effect of a preparation combating V. destructor depending on the number of mites found in a given colony. We chose this as a standard model of conduct that treats each bee colony as one organism subjected to individual parasite control. For this purpose, we created a bee colony with a mother-of-one breeding line and fertilised drones from one colony. Infection with V. destructor occurred naturally and uncontrollably. Without interfering with the colony's development, the frame insulator helped each colony's brood (mothers' reproductive potential) and the initial and final individuals from the mites themselves. The study was carried out in four species (two control species and two species with up to 20 and over 21 mites, respectively). Treatments with amitraz to combat damage were divided into four treatment subgroups: two treatments every four days or four treatments every two days. We observed the number of individuals that were protected in all subgroups in the average brood area. The reproductive potential of the sisters' mothers did not change after the treatments with amitraz, which indicated that amitraz did not affect the delegation of egg laying. The invasion rate was also tracked relative to the control group, which allowed us to conclude that a two-time treatment with amitraz reduced the frequency of mites and a four-time treatment checked the effectiveness. Tailoring the control of V. destructor in bee colonies may be an effective measure in the fight against this parasite.

Phylogenetic analysis of Starmerella apis in honey bees (Apis mellifera).

Honey bees are among the most effective pollinators that promote plant reproduction. Bees are highly active in the pollen collection season, which can lead to the transmission of selected pathogens between colonies. The clade Starmerella comprises yeasts that are isolated mainly from bees and their environment. When visiting plants, bees can come into contact with Starmerella spp. The aim of this study was to determine the prevalence and phylogenetic position of S. apis in bee colonies. Bee colonies were collected from nine apiaries in three regions. Ten colonies were sampled randomly from each apiary, and pooled samples were collected from the central part of the hive in each colony. A total of 90 (100%) bee colonies from nine apiaries were examined. Starmerella apis was detected in 31 (34.44%) samples, but related species were not identified. The 18S rRNA amplicon sequences of S. apis were compatible with the GenBank sequences of Starmerella spp. from India, Japan, Syria, Thailand, and the USA. The amplicon sequences of S. apis were also 99.06% homologous with the sequences deposited in GenBank under accession numbers JX515988 and NG067631.This is the first study to perform a phylogenetic analysis of S. apis in Polish honey bees.

Detection of Lotmaria passim and Crithidia mellificae in Selected Bumblebee Species.

Bumblebees (Bombus spp.) are an essential element of the ecosystem and the global economy. They are valued pollinators in many countries around the word. Unfortunately, there has been a decline in the bumblebee population, which is attributed to, among others, pathogens and reduced access to food due to the loss of natural nesting sites. Lotmaria passim and Crithidia mellificae, protozoan pathogens of the family Trypanosomatidae, commonly infect bumblebees, including in Poland. In this study, a Polish population of bumblebees was screened for L. passim and C. mellificae. The experiment was performed on 13 adult bumblebees belonging to 4 species: B. lapidarius, B. lucorum, B. pascuorum, and B. terrestris. Protozoa of the family Trypanosomatidae were identified by PCR. Only L. passim was identified in one B. pascuorum individual. Further research is needed to confirm the effect of concurrent pathogens on the decline of bumblebee populations.

Internal parasites infecting dogs in rural areas

Dogs which spend their lives in rural areas are primarily "sentinels" of animal herds. The dogs' close contact with humans and other animals obligates their caregivers to systematic deworming. The goal is to eliminate potential contamination with parasites which is mostly caused by direct contact or food and water contaminated with dogs' excretions. The aim of this study was to assess internal parasitic invasions on dogs from rural areas which spend most of their lives on farms. In total, 69 samples of fresh stools were collected for the analysis, including 26 from females and 43 from males. Coprological analysis of stool samples was accompanied with a survey where pet owners gave information on the number of visits to the veterinarian in suspicion of their dogs being infected with the parasites, the number of deworming treatments in the previous year and the dogs' behavior towards strangers. Also, the age, sex and body weight of dogs were recorded. Statistical analysis of the obtained results was performed using Statistica 12.5 with a medical set by Statsoft. In total, 29 (46,03%) out of 69 fecal samples contained developmental forms of parasites. Research has shown that animals most vulnerable to invasion were in the 6­10 years old age group, mostly males. In addition, the invasion of Toxocara canis roundworms occurred most frequently, as it was found in 12 (41.37%) animals (6 females and 6 males). The analysis of the questionnaires showed that the dog owners' knowledge of the risk of infection and parasites is unsatisfactory. This corresponded with a small number of deworming treatments. It is significant that nearly half of the owners never dewormed their dogs.

The influence of Dermanyssus gallinae and different lighting regimens on selected blood proteins, corticosterone levels and egg production in layer hens.

Egg production in battery cage systems in commercial poultry farms promotes uncontrolled growth of poultry ectoparasite Dermanyssus gallinae. Intermittent lighting regimens provided a promising alternative for controlling D. gallinae invasions. The study analysed the influence of D. gallinae invasions on selected blood protein fractions (albumin, α-, ß-, γ-globulin), corticosterone levels and egg production in Hy-Line Brown layer hens exposed to two lighting regimens: A (16 L:8D) and B (intermittent (4 L:2D); L-light, D-dark). Blood samples were collected from a total of 48 hens (divided into uninfested - UF, and infested - IF groups for each lighting regimen). The concentrations of protein fractions were analysed by electrophoresis on Cormay Gel Protein 100, and corticosterone levels were determined in a radioimmunoassay. The results of the study revealed concentrations of ß-globulin and corticosterone levels were significantly higher in IF than UF groups in both lighting regimens. However, both parameters were higher in hens exposed to lighting regimen B than lightening regimen A. Gamma-globulin concentrations were significantly lower in IF than UF groups in both lighting regimens. Egg production was significantly lower in all groups than commercial standard. D. gallinae and intermittent lightening regimen had interaction effect on the corticosterone level in hens. Strong decreasing (negative) linear relationship between corticosterone levels and egg production (r = -0.911) was reported.

Molecular analysis of cox-1 and 18S rRNA gene fragments of Eimeria species isolated from endangered grouse: capercaillie (Tetrao urogallus) and black grouse (Tetrao tetrix).

This paper is the first record describing the molecular analysis of Eimeria species occurring in capercaillie (Tetrao urogallus) and black grouse (Tetrao tetrix) which inhabit northern Eurasia and are species critically endangered of extinction. Actions undertaken to protect endangered species, such as breeding individuals in closed aviaries, could allow saving those birds, but they also pose risk of accidental healing of invasive diseases, like coccidiosis. Therefore, an investigation was conducted on fecal samples collected from the capercaillies and black grouse originating from the Kirov region (Russia) and breeding centers located in Poland. Results indicate that the average prevalence of Eimeria revealed 72% (average OPG = 3548) and 80% (average OPG = 5220) in capercaillies and black grouse respectively. Most of the Eimeria spp. oocysts were non-sporulated; however, two different morphological types were observed. The phylogenetic analysis of cox-1 and 18S rRNA genes revealed the analyzed Eimeria sequences to belong to two species. In addition, it showed some similarities between both analyzed genes. Most of the sequences obtained from both grouse species coccidia belonged to one species partially homologous to the Eimeria spp. isolated from ring-necked pheasant (approx. 94 and 96% for cox-1 and 18S rRNA genes, respectively). Two strains isolated from capercaillies imported from Russia were related to turkey coccidia: E. innocua and E. dispersa (97-99% homology) in the cox-1 gene analysis and only one of them was related to those Eimeria species in the 18S rRNA gene analysis (98-99% homology).

Characterisation of ail-positive Yersinia enterocolitica of different biotypes using HRMA.

Yersiniosis is one of the four most frequent foodborne zoonotic diseases in Europe, and Yersinia enterocolitica is the primary agent in human infections. The ail gene is an important chromosomal virulence marker of Y. enterocolitica which encodes Ail, a 17-kDa outer membrane protein that promotes attachment and invasion. In the present study, ail-positive Y. enterocolitica strains of different biotypes were examined using high resolution melting analysis (HRMA) and DNA sequencing. Genotype data relating to Y. enterocolitica strains isolated from different sources and belonging to different biotypes were compared. Applied method allowed efficient distinguishing of three genotypes and phylogenetic groups: 1A - included non-pathogenic Y. enterocolitica strains; 1B - consisted of highly pathogenic Y. enterocolitica strains and 2/4 - involved weakly pathogenic Y. enterocolitica strains. Amplicon genotyping based on HRMA supports rapid identification of ail SNPs correlated with biotype of examined Y. enterocolitica strains.

Preliminary studies on the occurrence of honeybee pathogens in the national bumblebee population

Dangerous pathogens that affect honeybees, including microsporidia of the genus Nosema, the acute bee paralysis virus (ABPV) and the deformed wing virus (DWV), also contribute to the global decline in bumblebee populations. In this study, a few insects with the bumblebee population present in country was checked for the presence of Nosema apis, Nosema bombi and Nosema ceranae microsporidia, ABPV, the chronic bee paralysis virus (CBPV) and DWV. Thirteen adult bumblebees (Bombus) representing 4 species were analyzed: Bombus lapidarius (2 workers, 2 males), Bombus lucorum (1 worker), Bombus pascuorum (5 workers, 1 male) and Bombus terrestris (2 workers). These specimens were found dead in two urban botanical gardens and four landscape parks in Lower Silesia. Microsporidia of the genus Nosema and CBPV were not detected in any of the examined samples. ABPV was identified in 2 insects of B. pascuorum and B. terrestris (2 workers). The presence of DWV was detected in 12 bumblebees (10 workers, 2 males).

A study of single nucleotide polymorphism in the ystB gene of Yersinia enterocolitica strains isolated from various wild animal species.

INTRODUCTION AND OBJECTIVE: Y. enterocolitica is the causative agent of yersiniosis. The objective of the article was a study of single nucleotide polymorphism in the ystB gene of Y. enterocolitica strains isolated from various wild animal species. MATERIALS AND METHOD: High-resolution melting (HRM) analysis was applied to identify single nucleotide polymorphism (SNP) of ystB gene fragments of 88 Y. enterocolitica biotype 1A strains isolated from wild boar, roe deer, red deer and wild ducks. RESULTS: HRM analysis revealed 14 different melting profiles - 4 of them were defined as regular genotypes (G1, G2, G3, G4), whereas 10 as variations. 24 of the examined Y. enterocolitica strains were classified as G1, 18 strains as a G2, 21 strains as a G3, and 15 strains as a G4. Nucleotide sequences classified as G1 revealed 100% similarity with the Y. enterocolitica D88145.1 sequence (NCBI). Analysis of G2 revealed one point mutation - transition T111A. One mutation was also found in G3, but SNP was placed in a different gene region - transition G193A. Two SNPs - transitions G92C and T111A - were identified in G4. Direct sequencing of 10 variations revealed 5 new variants of the ystB nucleotide sequence: V1 - transition G129A (3 strains); V2 - transitions T111A and G193A (2 strains); V3 - transitions C118T and G193A (1 strain); V4 - transitions C141A and G193A (2 strains); and V5 characterized by 19 SNPs: G83A, T93A, A109G, G114T, C116T, A123G, T134C, T142G, T144C, A150C, G162A, T165G, T170G, T174A, T177G, G178A, A179G, A184G and G193A (2 strains). The predominant genotype in isolates from wild ducks was G1; in red deer G2; in wild boar G3; in roe deer G1 and G4. CONCLUSIONS: The proposed HRM method could be used to analyze Y. enterocolitica biotype 1A strains isolated from different sources, including humans.

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.

Coccidia infections in homing pigeons of various age during the racing season.

Coccidiosis caused by Eimeria spp. is a common parasitic disease in homing pigeons. The study objective was to evaluate the incidence of coccidia infections in pigeon lofts during racing season. The intensity of coccidiosis was determined by floatation analyses of faeces samples collected from three pigeon groups performed in three replications (before the racing season, in mid-season and after the end of racing season). The presence of coccidia oocysts was determined in all faeces samples in each replication. At the end of the racing season, the average oocyst counts in faeces samples collected from pigeons that were flown for the first time increased by around 10% in relation to oocysts counts determined before the race. In flown pigeons (aged 2-4 years) a 2.5-9.9% drop was noted in oocysts counts subject to flock, whereas an increase of 15.7-17.3% was reported in parent flocks (unflown pigeons). The results of the experiment indicate that coccidia infections are a common problem in homing pigeons during racing season, which affects racing results and contributes to bird loss.

[The incidence of internal parasites in dogs and cats as dependent on the level of awareness among owners]. / Ocena zaleznosci zarazenia pasozytami wewnetrznymi psów i kotów od przygotowania hodowlano-weterynaryjnego wlascicieli.

The objective of the present study was to determine the incidence of internal parasites in dogs and cats owned by the employees of the Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, and by students and their families, as well as in pets presented to veterinary clinics and in animals from shelters. The study, conducted from January to May 2008, involved 135 dogs and 35 cats. Feces samples were analyzed for parasites by flotation technique with the use of Darling's solution, as described by Fülleborn. The highest percentage of parasite eggs was reported in feces samples collected from shelter dogs, including Toxocara and Toxascaris (9.1%), Uncinaria-Ancylostoma (12.1%) and Trichuris (12.1%), while in shelter cats the extensiveness of parasitic invasion was as follows: Toxocara cati--61.5%, Toxascaris--15.4%, Uncinaria-Ancylostoma--7.7%, Isospora spp.--7.7%. The lowest parasite incidence was observed among dogs and cats owned by veterinary students and members of their families. Of 20 examined animals, only one dog carried the eggs of Toxocara canis and one dog carried the eggs of Trichuris in their feces. Similar results were obtained in the group of pets owned by the employees of the Faculty of Veterinary Medicine: among 6 dogs and 13 cats examined in the study, Toxocara eggs were detected in one cat only (16.6%). In the group of pets presented to veterinary clinics, comprising 77 dogs and 8 cats, feces samples taken from 11 dogs contained the eggs of Toxocara canis, Toxascaris leonina, Uncinaria-Ancylostoma and Trichuris, while Isospora spp. oocysts were found in the stool of two cats.