Prevalence of selected tick-borne pathogens in wild ungulates and ticks in southern Spain.

A survey study was carried out to assess the occurrence of selected tick-borne pathogens (TBP) in wild ungulates in Mediterranean ecosystems in southern Spain. Spleen samples were collected from 1,132 wild ungulates, including 578 red deer, 269 wild boar, 135 mouflon, 121 fallow deer and 29 roe deer, between 2009 and 2015. Eighty-nine ticks collected from TBP-positive animals were also analysed. Samples were tested by PCR and sequenced whenever possible. TBP DNA was detected in 127 of 863 wild ruminants (14.7%; 95% CI: 12.4-17.3) including the following: Anaplasma phagocytophilum (9.2%), Babesia divergens (2.9%), Theileria sp. OT3 (1.7%), Borrelia afzelii (0.7%) and Theileria capreoli (0.2%), but no positive samples were detected in wild boar (0/269). All the strains from mouflon were identified as Theileria sp. OT3, while B. divergens and T. capreoli were mainly found in red deer. Co-infection with A. phagocytophilum and B. divergens, and A. phagocytophilum and Theileria spp. was detected in red deer and mouflon, respectively. The risk factor analysis showed that the prevalences of A. phagocytophilum and piroplasms were species-related. Eighty-nine tick specimens collected from ungulates found to be infected with the selected TBP were identified as Hyalomma lusitanicum (95.5%) and Ixodes ricinus (4.5%). Thirty ticks were positive for Anaplasma/Ehrlichia spp. (33.7%), 25 for Babesia/Theileria (28.1%) and two for B. burgdorferi s.l. (2.3%). Eleven specimens showed co-infections with Anaplasma/Ehrlichia and Babesia/Theileria (10.1%) or Anaplasma/Ehrlichia and B. burgdorferi s.l. (2.3%). The estimated prevalences obtained in the present study suggest the possible contribution of wild ruminants to the maintenance of some selected TBP in Mediterranean ecosystems in southern Spain, while the role of wild boar in the epidemiology of these pathogens seems to be limited in this region.

Effect of Bathing in a 0.1% Aqueous Solution of Ethacridine Lactate on Selected Physiological Parameters of Cornu Aspersum Müller Edible Snails.

INTRODUCTION: The prohibition of antibiotic use in edible snails obligates breeders to treat bacterial infections by different means, of which a common one is a bath in Gram-positive- and partially Gram-negative-bactericidal ethacridine lactate solution. The aim of the study was to determine the effect of bathing Cornu aspersum Müller snails in a 0.1% aqueous solution of ethacridine lactate on selected physiological parameters of haemolymph. MATERIAL AND METHODS: The study included 80 snails, divided into two equal groups (study and control). The study group was subjected to bathing in ethacridine lactate and the control group to bathing in tap water. Both groups were treated daily for seven days. The number of haemocytes in the haemolymph, the activity of alanine (ALT) and aspartate (AST) aminotransferases, and the concentration of urea were determined. RESULTS: In the study group, after exposure to ethacridine lactate solution an increase in ALT activity, changes in the De Ritis ratio, an increase in the amount of haemocytes, and a decrease in body weight were found. No such changes were detected in the control group snails or in animals after the first bath. CONCLUSION: Multiple applications of a 0.1% ethacridine lactate bath may adversely affect Cornu aspersum Müller snails.

Functionalization of 3D Chitinous Skeletal Scaffolds of Sponge Origin Using Silver Nanoparticles and Their Antibacterial Properties.

Chitin, as one of nature's most abundant structural polysaccharides, possesses worldwide, high industrial potential and a functionality that is topically pertinent. Nowadays, the metallization of naturally predesigned, 3D chitinous scaffolds originating from marine sponges is drawing focused attention. These invertebrates represent a unique, renewable source of specialized chitin due to their ability to grow under marine farming conditions. In this study, the development of composite material in the form of 3D chitin-based skeletal scaffolds covered with silver nanoparticles (AgNPs) and Ag-bromide is described for the first time. Additionally, the antibacterial properties of the obtained materials and their possible applications as a water filtration system are also investigated.

Wild animals as reservoirs of Anaplasma phagocytophilum for humans

Anaplasma phagocytophilum is a tick-transmitted obligate-intracellular gram-negative bacteria that causes emerging human zoonosis. A. phagocytophilum is transmitted by Ixodid ticks. Recent studies suggest that wild animals may be reservoirs of A. phagocytophilum for humans. The organism infects and survives within neutrophils. The infection diagnosis is based on the detection of morulae within granulocytes of peripheral blood, results of serological tests and detection of the DNA of A. phagocytophilum using specific polymerase chain reaction assays (PCR). A. phagocytophilum in most cases is transmitted to people by tick bites, but sometimes direct contact with infected blood may cause human granulocytic anaplasmosis (HGA). The possibility of infection should be taken into consideration at each occurrence of heavy disease symptoms after people come into contact with ticks.

Specificity of mass spectrometry (MALDI-TOF) in the diagnosis of Babesia canis regarding to other canine vector-borne diseases.

UNLABELLED: The canine vector-borne diseases (CVBD) is a term, which describes a range of infectious or/and parasitic diseases whose etiological agents are transmitted by vectors. CVBD are becoming more widely in the world in relation to global warming and the increasing number of infected vectors. The aim of this study was to assess rapid mass spectrometry (MS) - based proteomics analyses for diagnosis of Babesia canis, Anaplasma phagocytophilum and Borrelia burgdorferi infections in dogs. The study was conducted on four groups of dogs - healthy dogs (group 1, n=10) and dogs infected with B. canis (group 2, n=20), A. phagocytophilum (group 3, n=20) and B. burgdorferi (group 4, n=20) which demonstrated symptoms of the diseases. The MALDI-TOF (Matrix Assisted Laser Desorption Ionization with Time of Flight detector) MS technique revealed the presence of specific protein fraction of 51-52 kDa only in the blood serum of all the animals infected with the B. canis protozoa. The proteins are suspected to be disease markers, whereas the MALDI-TOF technique itself has high specificity and sensitivity and can be applied in the diagnosis of canine babesiosis. KEY WORDS: Anaplasma phagocytophilum, Babesia canis, Borrelia burgdorferi, MALDI-TOF, proteomics.

Detection of canine vector-borne diseases in eastern Poland by ELISA and PCR.

The aim of the study was to establish the prevalence of Ehrlichia canis, Anaplasma phagocytophilum and Borrelia burgdorferi in dogs in eastern Poland and to determine the factors associated with exposure (seroposity) or infection (PCR). Anti-A. phagocytophilum, anti-B. burgdorferi and anti-E. canis antibodies were determined in 400 dogs, using the SNAP 4Dx ® test (IDEXX Laboratories). In addition, PCRs were performed for the detection of E. canis, A. phagocytophilum and B. burgdorferi DNA. In reference to the risk factor analysis, a regression logistic model was determined for each aetiological agent. The overall seroprevalence was highest for B. burgdorferi (11.0 %), followed by A. phagocytophilum (8.0 %) and E. canis (1.5 %). Eleven healthy dogs were found to be infected with A. phagocytophilum, as determined by PCR, while the remainder were seronegative. For B. burgdorferi, the DNA of the spirochetes was detected in the blood of 20 dogs, while the presence of anti-B. burgdorferi IgG was detected in the sera of ten of these. For E. canis, none of the dogs tested positive by PCR. Tick control was included as a protective factor for A. phagocytophilum and B. burgdorferi, while the origin (rural) was included as a risk factor for B. burgdorferi and A. phagocytophilum infection. In addition, breed (pure) was a risk factor for B. burgdorferi infection, and sex (female) was a risk factor for E. canis.

The occurrence of Anaplasma phagocyto- philum in wild bison from the Bialowieza Primeval Forest in Eastern Poland.

The aim of the present study was to investigate the occurrence of Anaplasma spp. in a group of 120 wild bison (Bison bonasus) from the Bialowieza Primeval Forest in eastern Poland and to determine which species of Anaplasma could infect these animals based on a PCR of a part of the 16S rRNA gene followed by sequencing. The PCR technique showed the presence of 16S rRNA Anaplasma spp. genetic material in the blood of 22 from a total of 120 animals. DNA amplification by means of the primers EHR 521 and EHR 747 gave a product size of 252-bp. The sequences of the PCR products obtained showed 100% homology with each other and 100% homology with the Anaplasma phagocytophilum GU 183908 sequence from our earlier study, isolated from a horse with a clinical case of anaplasmosis. The similarity of the sequences with the sequences of the 16S rRNA gene isolated from various Anaplasma species deposited in the GeneBank, ranged between 95.8% and 98.8%. Based on the results of molecular analysis, bacterial DNA detected in the blood of 22 wild bison was identified as Anaplasma phagocytophilum.

Equine granulocytic anaplasmosis.

Anaplasma phagocytophilum is an emerging pathogen of horses that is transmitted by Ixodid ticks. Recent studies suggest that multiple strains of A. phagocytophilum may be circulating in wild and domestic animal populations, and these strains may have differential host tropisms and pathogenicity. The organism infects and survives within neutrophils. Co-infections with other tick-borne pathogens may occur, especially Borrelia burgdorferi. A. phagocytophilum causes an acute febrile illness in horses with lethargy, inappetence, lameness and hemorrhages. Diagnosis is based on finding morulae within granulocytes in the peripheral blood, and detection of the DNA of A. phagocytophilum using specific polymerase chain reaction assays. Most reports suggesting that anaplasmosis is a self-limiting disease that responds well to a tetracycline therapy.

Comparative analysis of 16S RNA nucleotide sequences of Anaplasma phagocytophilum detected in the blood of horses from various parts of Europe.

The aim of this study was to conduct a comparative analysis of the 16S rRNA gene fragment nucleotide sequences for Anaplasma phagocytophilum strains detected in the blood of horses from various parts of Europe. The study comprised 234 horses that had had contact with ticks. Using PCR, the genetic material of A. phagocytophilum was identified in the blood of 42 animals. The sequences of the 16S RNA gene amplicons that were obtained from our A. phagocytophilum isolates had 100 % similarity with each other and 96.4-100 % similarity with Anaplasma spp. sequences selected from those available in GenBank. Nucleotide substitutions at positions 248 and 249 were demonstrated in all the 16S RNA gene sequences of Anaplasma obtained in our study. This may indicate the emergence of a new rickettsial genotype that is the cause of equine granulocytic anaplasmosis in southern and eastern Europe. These results add new information on the epidemiology and genetic diversity of A. phagocytophilum detected in the blood of horses from southern and eastern Europe.