Vector-borne parasites in dogs from Ukraine translocated to Poland following Russian invasion in 2022.

INTRODUCTION: Since 24 February 2022, the day the Russian aggression against Ukraine began, millions of refugees and thousands of pets crossed the Polish-Ukrainian border. Additionally, an unknown number of shelter and stray dogs and cats were rescued and translocated to Poland by private persons and non-profit organizations. The aim of the present study was to examine rescued dogs and cats for presence of canine vector-borne parasites to determine the role of armed conflict in spreading these parasites. METHODS: In July 2022 blood samples were collected from two animal shelters in central Poland hosting dogs and cats rescued from Ukraine. Animals were imported from various regions of Ukraine, including eastern and southeastern Ukraine (military conflict area). Fifty-three dogs (51 from two shelters and two owned ones) and one shelter cat were examined by molecular methods (PCR and sequencing) for the presence of Babesia/Theileria spp., Dirofilaria spp. and Hepatozoon spp. DNA. RESULTS: We detected Dirofilaria immitis, a parasite species non-endemic in Poland, in two dogs translocated from Ukraine (2/53 = 3.8% [95% CI 0.5-13.0%]). One dog had a history of previous heartworm infection. High prevalence of Hepatozoon canis (51% [95% CI 36.8-64.9%]) was noted among translocated dogs. Prevalence of Dirofilaria repens in Ukrainian dogs (18.9% [95% CI 9.4-32.0%) was similar to prevalence in dogs from central Poland (12%). Co-infection of D. repens and D. immitis was found in two dogs and six dogs were co-infected by D. repens and H. canis. Hepatozoon canis infection was also identified in an imported cat. CONCLUSION: We confirmed that this military conflict has facilitated the spread of canine vector-borne parasites, including zoonotic species.

Comparison of automatic methods MALDI-TOF, VITEK2 and manual methods for the identification of intestinal microbial communities on the example of samples from alpacas (Vicugna pacos).

Introduction: Universally, in microbiological diagnostics the detection of live bacteria is essential. Rapid identification of pathogens enables appropriate remedial measures to be taken. The identification of many bacteria simultaneously facilitates the determination of the characteristics of the accompanying microbiota and/or the microbiological complexity of a given environment. Material and Methods: The effectiveness of the VITEK2 Compact automated microbial identification system and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS), analytical profile index (API) and Remel RapID tests were compared in identification of bacteria isolated from the alpaca gastrointestinal tract. Results: Most isolates were Gram-positive, such as Bacillus cereus, Bacillus flexus, Bacillus licheniformis, Bacillus pumilus and Bacillus subtilis; Enterococcus faecium, Enterococcus gallinarum, Enterococcus hirae and Enterococcus casseliflavus; Staphylococcus aureus, Staphylococcus equorum, Staphylococcus lentus, Staphylococcus pseudintermedius and Staphylococcus sciuri; Paenibacillus amylolyticus; Cellulosimicrobium cellulans; Leuconostoc mesenteroides; Clostridium perfringens; Corynebacterium stationis, Corynebacterium xerosis, and Corynebacterium diphtheriae (the last only isolated manually by API Coryne and the VITEK2 system and Corynebacteria (CBC) card). Corynebacterium diphtheriae was misidentified by MALDI-TOF MS as Candida lipolytica (currently Yarrowia lipolytica). Gram-positive and Gram-variable Micrococcus luteus were also isolated. Gram-negative Enterobacter cloacae, Enterobacter gergoviae, Enterobacter hormaechei and Enterobacter ludwigii; E. coli; Klebsiella pneumoniae subsp. pneumoniae; Citrobacter braakii and Citrobacter freundii; Serratia liquefaciens, Serratia odorifera and Serratia marcescens; Morganella morganii subsp. morganii; Providencia alcalifaciens; Pseudomonas aeruginosa; Stenotrophomonas maltophilia; Moraxella osloensis; and Ochrobactrum intermedium were also found. The yeasts Candida albicans, Candida haemulonii and Candida ciferrii were also present. Conclusion: MALDI-TOF MS enabled the identification of pathogens and opportunistic pathogens from the alpaca gut which may represent a high risk to human and animal health.

Occurrence of Dirofilaria repens in wild carnivores in Poland.

Dirofilaria repens is an expanding vector-borne zoonotic parasite of canines and other carnivores. Sub-clinically infected dogs constitute the most important reservoir of the parasite and the source of infection for its mosquito vectors. However, occurrence of D. repens infection in wild animals may contribute to the transmission of the parasite to humans and may explain the endemicity of filariae in newly invaded regions. The aim of the current study was to determine the occurrence of D. repens in 511 blood and spleen samples from seven species of wild carnivores (wolves, red foxes, Eurasian badgers, raccoons, raccoon dogs, stone martens, and pine martens) from different regions of Poland by means of a PCR protocol targeting the 12S rDNA gene. Dirofilaria repens-positive hosts were identified in seven of fourteen voivodeships in four of the seven regions of Poland: Masovia, Lesser Poland, Pomerania and Warmia-Masuria. The highest prevalence was found in Masovia region (8%), coinciding with the highest previously recorded prevalence in dogs in Central Poland. The DNA of Dirofilaria was detected in 16 samples of three species (total prevalence 3.13%). A low and similar percentage of positive samples (1.9%, 4.2% and 4.8%) was recorded among badgers, red foxes, and wolves, respectively. Dirofilaria repens-positive hosts were identified in seven of fourteen voivodships. Based on detection in different voivodeships, D. repens-positive animals were recorded in four out of the seven regions of Poland: in Masovia, Lesser Poland, Pomerania, and Warmia-Masuria. The highest prevalence of filariae was found in Masovia region (8%), reflecting the highest previously recorded prevalence in dogs (12-50%) in Central Poland. In summary, we conducted the first comprehensive study on the epidemiology of D. repens in seven species of wild hosts in all seven regions of Poland and identified the first case of D. repens infection in Eurasian badgers in Poland and the second in Europe.

Haematological indices and immune response profiles in dogs naturally infected and co-infected with Dirofilaria repens and Babesia canis.

Co-infections with Dirofilaria repens and Babesia canis are rarely reported in the literature and there is very limited knowledge of their impact on canine health. Central Poland is endemic for both parasites, posing a risk of co-infections in dogs. To evaluate the impact of co-infection with B. canis and D. repens on canine health, four groups of dogs were examined: healthy dogs, dogs infected with B. canis, dogs infected with D. repens and dogs co-infected with both species. Blood parameters indicative of anaemia, kidney and liver damage were analysed statistically. Additionally, expression levels of immune response genes were quantified and compared, to define the type of immune response typically encountered in single- and co-infections. In dogs infected with D. repens, no major alterations in blood parameters were observed. Dogs infected with B. canis suffered from anaemia, kidney and liver insufficiency. In contrast, dogs co-infected with D. repens and B. canis showed milder alternation in blood biochemical parameters associated with liver (ALP activity) and kidney (serum urea and creatinine levels) dysfunction, compared to dogs infected only with B. canis. The expression of genes associated with cellular (Th1-mediated) (STAT4 and INF-γ), humoral (Th2-mediated) (STAT6, GATA3, SOCS3, IL-13) and regulatory (IL-10) responses was quantified. For this analysis, dogs infected with B. canis were divided into two groups-'Babesia 1' (mild babesiosis), 'Babesia 2' (severe babesiosis). All the tested factors, except INF-γ, were found to be expressed in dogs infected with D. repens. In 'Babesia 1' dogs, expression of GATA3 was highest, while in 'Babesia 2'-INF-γ and SOCS3 dominated. IL-13 expression was predominant in dogs infected with D. repens, and STAT6 and IL-10 predominated in dogs with co-infections.

Tick-Borne Pathogens, Babesia spp. and Borrelia burgdorferi s.l., in Sled and Companion Dogs from Central and North-Eastern Europe.

Ticks are important vectors of numerous pathogens of medical and veterinary significance. The aim of the current study was to determine the prevalence of Babesia spp. and Borrelia burgdorferi s.l. in sled and pet dogs from Central and North-Eastern Europe. Neither Babesia spp. nor Borrelia burgdorferi s.l. infections were detected in sled dogs from seven countries (Poland, Lithuania, Latvia, Estonia, Belarus, Russia and Finland). The DNA of Babesia spp. was detected in 100% of symptomatic and 5.4% of asymptomatic pet dogs from Poland. Similarly, the DNA of Babesia spp. was identified in 82% of symptomatic and 3.8% of asymptomatic pet dogs from Ukraine. The DNA of Borrelia burgdorferi s.l. was detected in 4.4% of pet dogs. Molecular typing confirmed the presence of Babesia canis and Borrelia burgdorferi sensu stricto (s.s.) in selected samples. Four dogs were co-infected by B. canis and Borrelia burgdorferi s.l. Tick-borne pathogens constitute a serious health threat to pet dogs in Central and South-Eastern Europe, but were not observed among sled dogs from the same region of Europe nor in the Baltic countries.

Disorders in Blood Circulation As a Probable Cause of Death in Dogs Infected with Babesia Canis.

INTRODUCTION: The purpose of the study was to investigate post-mortem changes in dogs infected with Babesia canis and to establish the probable cause of death of the affected animals. MATERIAL AND METHODS: Cadavers of six dogs that did not survive babesiosis were collected. Necropsies were performed and samples of various organs were collected for histological examination. RESULTS: Necropsies and histological examinations revealed congestion and oedemata in various organs. Most of the dogs had ascites, hydrothorax or hydropericardium, pulmonary oedema, pulmonary, renal, hepatic, and cerebral congestion, and necrosis of cardiomyocytes. CONCLUSION: These results suggested disorders in blood circulation as the most probable cause of death. However, the pulmonary inflammatory response and cerebral babesiosis observed in some of these dogs could also be considered possible causes of death. This study also showed a possible role for renal congestion in the development of renal hypoxia and azotaemia in canine babesiosis.

Monitoring the expansion of Dermacentor reticulatus and occurrence of canine babesiosis in Poland in 2016-2018.

BACKGROUND: The significance of tick-borne diseases has increased considerably in recent years. Because of the unique distribution of the tick species Dermacentor reticulatus in Poland, comprising two expanding populations, Eastern and Western that are separated by a Dermacentor-free zone, it is important to conduct studies on the process of tick expansion and emergence of canine babesiosis. The main aim of the current study was to monitor the expansion of D. reticulatus populations from spring 2016 to autumn 2018 to determine (1) the actual geographical range of this tick species, and (2) and the seasonal/annual shift in range limits and changes in distance between Western and Eastern populations of ticks (the size of the non-endemic area). METHODS: Ticks were collected in spring/autumn during a 3-year study. From each season and year at least three pairs of sites from the Western and Eastern populations were selected. Then the mean distance between paired sites was calculated for each season and year. We collected and analyzed data from veterinary clinics on the number of canine babesiosis cases treated in the clinic during a whole year (2018). RESULTS: Accordingly, further expansion of the two D. reticulatus populations was recorded, mainly along river basins. Marked colonization of the gap zone was observed, with a mean annual shift in the range of 2.5-10 km and a steadily decreasing distance between the two tick populations. The occurrence of babesiosis in different regions revealed low numbers of cases in Western Poland (19 cases/year) and the gap area (only 7 cases/year) and high incidence (up to 250 cases/1000 dogs) and fatality (total 3.65%) in Central and Eastern Poland. Strong associations were found geographically between tick and babesiosis occurrence and temporally in the seasonal patterns of occurrence of ticks and outbreaks of babesiosis. CONCLUSIONS: We documented the shift in range limits and continued process of colonization of the gap zone accompanied by the emergence of canine babesiosis in the Eastern expansion zone. Updated maps of the distribution of ticks and occurrence of babesiosis in different regions of Poland have allowed us to predict of the emergence of pathogens vectored by D. reticulatus. Incidence (per 1000 dogs) of canine babesiosis in veterinary clinics by current range of D. reticulatus.

Impact of Babesia microti infection on the initiation and course of pregnancy in BALB/c mice.

BACKGROUND: Protozoa in the genus Babesia are transmitted to humans through tick bites and cause babesiosis, a malaria-like illness. Vertical transmission of Babesia spp. has been reported in mammals; however, the exact timing and mechanisms involved are not currently known. The aims of this study were to evaluate the success of vertical transmission of B. microti in female mice infected before pregnancy (mated during the acute or chronic phases of Babesia infection) and that of pregnant mice infected during early and advanced pregnancy; to evaluate the possible influence of pregnancy on the course of parasite infections (parasitaemia); and to assess pathological changes induced by parasitic infection. METHODS: The first set of experiments involved two groups of female mice infected with B. microti before mating, and inseminated on the 7th day and after the 40th day post infection. A second set of experiments involved female mice infected with B. microti during pregnancy, on the 4th and 12th days of pregnancy. Blood smears and PCR targeting the 559 bp 18S rRNA gene fragment were used for the detection of B. microti. Pathology was assessed histologically. RESULTS: Successful development of pregnancy was recorded only in females mated during the chronic phase of infection. The success of vertical transmission of B. microti in this group was 63%. No evidence of pregnancy was found in females mated during the acute phase of infection or on the 4th day of pregnancy. In the group infected on the 12th day of pregnancy, numerous complications including loss of pregnancy and stillbirths were recorded. During the acute phase of infection, parasitaemia was lower in pregnant females in comparison to infected, non-pregnant control females. CONCLUSIONS: Acute B. microti infection prevents the initiation of pregnancy and embryonic development if it occurs during the first trimester, and causes severe complications in foetal BALB/c mice in the second and third trimesters of pregnancy. Chronic B. microti infection has no detrimental impact on the initiation and development of pregnancy, but results in congenital infection of the offspring. Further study is required to determine the extent to which maternal anti-babesial immune responses contribute to compromise pregnancy in the murine model of congenital Babesia infection.

Emerging risk of Dirofilaria spp. infection in Northeastern Europe: high prevalence of Dirofilaria repens in sled dog kennels from the Baltic countries.

Dirofilariasis is a fast-spreading disease of dogs and humans in Europe. We investigated whether Dirofilaria spp. have spread northwards in Europe, invading the Baltic countries. Altogether, 424 blood samples were collected from eight countries in the period 2017-2019, including 227 samples from sled dogs and 197 samples from other dogs. PCR amplification and sequencing were conducted employing three genetic markers (mitochondrial [mt] 12S rDNA, mt cytochrome c oxidase subunit I [COI] gene and mt dehydrogenase subunit I [NAD1] gene). The SNAP test (IDEXX) for detection of D. immitis infections was also implemented. The DNA of D. repens was detected in 59 of 424 dogs (prevalence 13.9%). D. repens was found in sled dogs from Lithuania, Latvia, Poland and Belarus. Only one dog from Estonia was infected, apparently an imported case. The highest prevalence was recorded in Lithuania (38%). Among pet dogs from the Ukraine, six dogs tested positive (3.8%). Our study has revealed a high prevalence of D. repens infections in Lithuania and Latvia, but no evidence for spread of the heartworm D. immitis. We conclude that sled dog kennels constitute hot spots for D. repens transmission.

Abundance of the tick Dermacentor reticulatus in an ecosystem of abandoned meadows: Experimental intervention and the critical importance of mowing.

The effect of agricultural activities on the environment has been falling in many areas of Europe in recent years and the associated abandonment of crop fields, meadows and pastures may enable an increase in tick densities. In the present study we assessed whether regular mowing would have a negative effect on Dermacentor reticulatus populations and whether the cessation of regular mowing would cause an increase in abundance of D. reticulatus ticks. Two field experiments were conducted during a five-year period (2012-2016) in the Mazowieckie (Mazovia) region of Central Poland. Experiment 1: The long-term effect of mowing on tick population was tested in the meadow ecosystem of Stoski, an old fallow land plot that was mowed three times a year. Experiment 2: Neglecting the cultivation by abandonment of arable land was evaluated in Kury village. Four areas (2 experimental 'fallow lands', 2 control meadows) were selected. The first fallow land plot was a fenced off area comprising mostly of a horse pasture and the second fallow land plot was designated in an old abandoned orchard. At each site, ticks were collected in consecutive springs and autumns by dragging at least twice during each season from experimental and control areas. Altogether 1452 D. reticulatus ticks were collected and their densities were compared by multifactorial ANOVA. In the end of the first experiment, a significant decrease (6 times) in tick abundance was observed in the mowed area in comparison to old fallow land. In the end of the second experiment, tick abundance was three times higher in the experimental fallow lands in comparison to the control meadows. In conclusion it was found that regular mowing significantly reduced the density of questing D. reticulatus ticks in open areas. Cessation of mowing may enhance the number of ticks and the associated risk of acquiring tick-borne diseases.

Origins of recently emerged foci of the tick Dermacentor reticulatus in central Europe inferred from molecular markers.

The ornate dog tick Dermacentor reticulatus is vector of several blood parasites, including Babesia canis, a causative agent of babesiosis. The geographical range of D. reticulatus in Europe is discontinuous with a gap separating eastern and western macroregions. New foci observed in several locations in western and central Europe were considered an expansion of the western population, including foci in western Poland. In the present paper we used molecular markers to identify the origins of these foci, and we compared their genetic polymorphism to D. reticulatus collected in sites situated within the eastern population. The overall polymorphism in mt 16S rDNA was low, and all sites from the western population shared the same haplotype suggesting the expansion in this area. In the marker 5.8S rDNA-ITS2 we found no differences in polymorphism between sites from eastern Poland (eastern population), and newly emerged foci in western Poland considered a putative expansion zone of the western population. However, the sites from western Poland differed considerably from nearby German site. Our results show that foci in western Poland could not have originated from D. reticulatus from the western population, as previously thought. We found that the state border following river hinders considerably gene flow between adjacent sites what suggest that natural dispersal of D. reticulatus by wildlife is unlikely, and the emergence of new foci should rather be contributed to human-associated dispersal. We propose that livestock, and pets travelling with their owners are the most probable source of new foci, and they can easily transfer ticks within a country but not between countries.

The prevalence of Dirofilaria repens in cats, healthy dogs and dogs with concurrent babesiosis in an expansion zone in central Europe.

BACKGROUND: Dirofilaria repens is a mosquito-transmitted, filarial nematode parasitizing dogs, cats and other carnivores. Recently, this parasite has spread in central Europe, including Poland. The aim of the present study was to estimate the prevalence of D. repens in cats and dogs in different regions of the country and to investigate the occurrence and consequences of co-infection with another fast-spreading vector-borne parasite, Babesia canis. RESULTS: In the period 2013-2015, 147 blood samples from cats from central Poland and 257 blood samples from dogs from central, northern, southern and western Poland were collected. Prevalence of D. repens was determined by amplification and sequencing of the 12S rDNA gene fragment. Among dogs, 94 samples originated from clinically healthy dogs from central Poland (Masovia) and 58 samples originated from dogs that were infected with B. canis. Prevalence of D. repens was compared between these two groups of dogs. For the first time D. repens was identified in a cat from central Europe (0.7 % [95 % CL: 0-4.1 %]). The DNA of the filarial endosymbiotic bacterium Wolbachia was detected in two cats (1.4 % [95 % CL: 0-5.5 %]). In dogs, the parasite was detected only in samples from central Poland (Masovia) (local prevalence = 38 % [95 % CL: 25.9-51.8 %]). Prevalence of D. repens was significantly higher in dogs with babesiosis (90 % [95 % CL: 81.6-94.5 %]). Co-infections of D. repens and B. canis were confirmed by sequencing in 30 dogs with babesiosis, but no co-infections were identified in healthy dogs from Masovia. Statistical analyses of blood parameters revealed that dogs with co-infections suffered more severe anemia and thrombocytopenia, but presented milder changes in biochemical parameters (i.e. less elevated concentration of alkaline phosphatase [ALP] and serum urea) suggesting lower risk of hepatic or renal failure in comparison to dogs infected only with B. canis. CONCLUSIONS: These findings are important due to the spread of dirofilariosis and babesiosis in central Europe, as microfilaraemic dogs seem to be more prone to babesiosis. The possible protective effect of the nematode infection against hepatic or renal failure in canine babesiosis and its mechanisms require further investigations.

The first evidence for vertical transmission of Babesia canis in a litter of Central Asian Shepherd dogs.

INTRODUCTION AND OBJECTIVE: Tick-borne infections constitute an increasing health problem in dogs and may lead to death, especially in young or elderly individuals. Canine babesiosis constitutes a serious health problem in dogs worldwide. The aim of the study was to verify the probability of vertical transmission of Babesia canis between the bitch and the pups. MATERIALS AND METHODS: In Autumn 2011, cases of babesiosis were diagnosed in a litter of 6-week-old puppies of a Central Asian Shepherd dog. Immediately following the first case of infection, blood samples were collected from all the pups in the litter (n=10) and from the female. Detection of Babesia infection was performed by molecular and microscopical techniques. RESULTS: The presence of B. canis DNA was detected using PCR in three pups, presenting at the time or 24-48 hours later with babesiosis symptoms, and in their asymptomatic mother. The isolates derived from the pups and the female - 520 bp 18S rRNA gene fragment - were compared and analyzed. All isolates from the pups and their mother were identical and showed 100% homology with B. canis group B (EU622793), supporting the same source of infection. Additionally, the USG of the peritoneal cavity was performed in the female, presenting evidence for splenomegaly. CONCLUSIONS: On the basis of (1) the same timing of three pup cases; (2) the identical B. canis sequences derived from all positive dogs; (3) evident splenomegaly in the asymptomatic female, this provides the first evidence of the vertical transmission of this piroplasm in dogs.

The risk of vector-borne infections in sled dogs associated with existing and new endemic areas in Poland. Part 2: Occurrence and control of babesiosis in a sled dog kennel during a 13-year-long period.

The achievements of sled dogs in competitions depend both on their training and on their health. Vector-borne infections may lead to anemia, affect joints or heart muscles or even cause death. Canine babesiosis is an emerging, quickly spreading tick-borne disease in Central Europe. Over a 13-year period (2000-2012) the occurrence of babesiosis cases was analyzed in one sled dog kennel situated in Kury, a village near Tluszcz (N 52°24'56.78″, E 21°30'37.55″) in Central Poland. Twenty cases/episodes of babesiosis were noted among the 10-12 dogs living in the kennel. In 2000-2004, no cases of babesiosis were noted; the first two cases were noted in April 2005. Since that time, only one dog remained uninfected; 6 dogs were infected once, 3 dogs demonstrated symptoms of babesiosis twice, one dog was infected three times and one dog had it five times. Babesiosis appeared in Spring and Autumn, despite the application of anti-tick treatment. No fatal cases were recorded, but in one case a splenectomy was performed due to splenomegaly and spleen rupture. Additionally, the abundance of the main Babesia canis vector, the Dermacentor reticulatus tick, was estimated and monitored during a 4-year period (2008-2012) close to the dog kennel. The abundance of questing ticks was high in 2008 and 2009, but dropped by 10-fold between 2010 and 2012, when the abandoned meadow was cut and used as horse pasture by the local farmer. The regular occurrence, typical seasonal pattern and identification of B. canis DNA in questing tick from this locality confirmed the establishment of a new hyper enzootic region for canine babesiosis. The effectiveness and schedule of applied preventive measures were discussed.

The risk of vector-borne infections in sled dogs associated with existing and new endemic areas in Poland: Part 1: A population study on sled dogs during the racing season.

The achievements of sled dogs in competitions depend both on their training and on their health. Vector-borne infections may lead to anaemia, affect joints or heart muscle or even cause death. Between December 2009 and October 2010, one hundred and twenty six individual blood samples were collected from 26 sled dog kennels situated in different regions of Poland. The majority of samples were taken during the racing season (winter 2009/10). The prevalences of 3 vector-borne infections- including 2 'old pathogens' Anaplasma phagocytophilum and Babesia canis, and 'new pathogen' Hepatozoon canis-were estimated in sled dogs using PCR and nested PCR. Additionally, 25 serum samples originating from a subset of 3 kennels situated in a tick-borne encephalitis (TBE) endemic area (Mazowiecki region), were tested for antibodies against the tick-borne encephalitis virus (TBEV). Because of the recently reported occurrence of Dirofilaria repens in Central Poland and that of fatal cases of unknown aetiology in two of the kennels, blood samples collected from dogs at these kennels in 2010 and in February-May 2013 and from two unaffected kennels were checked for evidence of presence of this parasite. Babesia canis DNA was detected in 11 sled dogs (4 with clinical babesiosis, 7 asymptomatic; 8.7%) inhabiting mainly endemic regions of Poland (9/11 cases). Three serum samples originating from one location tested positive for TBEV antibodies (total seroprevalence: 3/25=12%, local seroprevalence: 3/12=25%). The risk of TBEV infection was associated with previous B. canis infections. Dirofilaria repens DNA was detected in 15 dogs (44%). Prevalence was especially high in two sled dog kennels situated near Grodzisk Mazowiecki (50-57%). No blood samples tested positive for A. phagocytophilum or H. canis DNA. The present study has established that the prevalence of vector-borne pathogens in working sled dogs is significant in the endemic regions and has justified the important role of surveillance of reservoir hosts in the epidemiology of TBE. Our results emphasize the need for regular monitoring for the presence of D. repens.

Babesia canis and tick-borne encephalitis virus (TBEV) co-infection in a sled dog.

INTRODUCTION AND OBJECTIVES: Sporting dogs, including sled dogs, are particularly prone to tick-borne infection either due to training/racing in forest areas or through visits to endemic areas. The aim was to present tick-borne infections in a 6-dog racing team after a race in Estonia. MATERIALS AND METHODS: On the 4th day after return to Poland, the first dog presented with babesiosis symptoms and was diagnosed and treated accordingly. Next morning, the dog showed neurological symptoms and was diagnosed with tick-borne encephalitis (TBE). Diagnosis was confirmed by a high level of IgG antibodies (922 IU/ml), detected in serum 3 months later. The second dog presented with babesiosis symptoms on the 7th day after return. Babesia DNA was extracted from blood, amplified and sequenced to answer the question of whether the dogs became infected during the race in Estonia or in Poland. RESULTS AND CONCLUSIONS: Sequencing of a fragment of Babesia 18S rDNA revealed that these two isolates were identical to one another and closely related to the B. canis sequence originally isolated from the dog and Dermacentor reticulatus ticks in Poland. Thus, this is the first confirmed case of B.canis and TBEV co-infection and first confirmed case of TBE in a dog in Poland.

Spatiotemporal characterization of mTOR kinase activity following kainic acid induced status epilepticus and analysis of rat brain response to chronic rapamycin treatment.

Mammalian target of rapamycin (mTOR) is a protein kinase that senses nutrient availability, trophic factors support, cellular energy level, cellular stress, and neurotransmitters and adjusts cellular metabolism accordingly. Adequate mTOR activity is needed for development as well as proper physiology of mature neurons. Consequently, changes in mTOR activity are often observed in neuropathology. Recently, several groups reported that seizures increase mammalian target of rapamycin (mTOR) kinase activity, and such increased activity in genetic models can contribute to spontaneous seizures. However, the current knowledge about the spatiotemporal pattern of mTOR activation induced by proconvulsive agents is rather rudimentary. Also consequences of insufficient mTOR activity on a status epilepticus are poorly understood. Here, we systematically investigated these two issues. We showed that mTOR signaling was activated by kainic acid (KA)-induced status epilepticus through several brain areas, including the hippocampus and cortex as well as revealed two waves of mTOR activation: an early wave (2 h) that occurs in neurons and a late wave that predominantly occurs in astrocytes. Unexpectedly, we found that pretreatment with rapamycin, a potent mTOR inhibitor, gradually (i) sensitized animals to KA treatment and (ii) induced gross anatomical changes in the brain.

Risk factors and control of intestinal parasite infections in sled dogs in Poland.

Training and racing constitute serious challenges for working sled dogs. Attainment of the highest levels of stamina and speed are possible only by completely healthy dogs. Infections with nematodes as whipworm Trichuris sp. or hookworms Uncinaria/Ancylostoma can significantly reduce the fitness of working dogs leading to anemia or even to death. In the middle of the racing season, between December 2009 and April 2010, 108 individual fecal samples were collected from 25 sled dog kennels situated in different regions of Poland. Saturated salt flotation was performed for helminth egg detection. The immunofluorescent assay MeriFluor Cryptosporidium/Giardia and nested PCRs on 18S rRNA (Cryptosporidium spp.) and TPI gene (Giardia spp.) were carried out for detection of intestinal protozoa. Overall prevalence of 6 species of intestinal parasites was 68% in sled dogs (73/108). In 51 samples the eggs of a single species of helminth were detected (47%), two nematode species were detected in 13%, three species of nematodes were found in two dogs. The most prevalent helminths were the hookworms Uncinaria/Ancylostoma-identified in 36% of kennels, and in 34% of sled dogs. Toxocara eggs were detected in 36% of kennels, in 17% of dogs. Trichuris sp. eggs were found in 20% of kennels (5/25), in 13% of dogs. Cysts/oocysts of intestinal protozoa were detected in 31% of sled dogs. The most prevalent was Giardia spp. infection-in 54% of kennels [13/24], in 28% of dogs. Cryptosporidium spp. infections were identified in 37.5% of kennels [9/24], in 13% of dogs. Two sequenced Giardia isolates presented 100% homology with G. intestinalis Assemblage C isolate (AY228641.1), specific for dogs. A range of factors was shown to affect the prevalence of intestinal parasites in sled dogs. The highest prevalence of parasites was found among dogs from large kennels (housing >3 dogs), in dogs less than 2 years old, and in kennels, where prophylactic treatment was carried out 1-4 times a year. The present study has demonstrated a high prevalence of intestinal parasites in working sled dogs in Poland, including the zoonotic human pathogens Toxocara or Cryptosporidium.

Babesia canis and other tick-borne infections in dogs in Central Poland.

Vector-borne infections constitute increasing health problem in dogs worldwide, including sled dogs, dramatically decreasing the fitness of working dogs and even leading to death. In the period 2006-2008 eighty-two blood samples were collected from eight sled dog kennels in Central Poland. The prevalence of four vector-borne infections (Babesia canis, Bartonella sp., Anaplasma/Ehrlichia and Borrelia burgdorferi) was estimated in 82 sled dogs using PCR and nested PCR for diagnosis and the same methods were used to identify the vector-borne pathogens in 26 dogs presenting at veterinary clinics with symptoms of vector-borne diseases. None of four studied vector-borne pathogens was detected in samples originating from veterinary clinics. Among the remaining 82 dogs B. canis infections were confirmed in three dogs undergoing treatment for babesiosis. The DNA of tick-borne pathogens was also found among 22 (27.8%) of the 79 apparently healthy dogs, including 20 cases of B. canis infection (25.3%), one case of B. burgdorferi s.l. infection and one case of Anaplasma phagocytophilum infection. No evidence of Bartonella spp. and Ehrlichia canis infections were found in this set of samples. Sequencing of a Babesia fragment of 18S rDNA amplified from acute (n=5) and asymptomatic (n=5) cases revealed that all isolates were identical to the Babesia canis canis sequence, originally isolated from Dermacentor reticulatus ticks in Poland. A range of factors was shown to affect the distribution of babesiosis in sled dogs. The data are also discussed in respect to the health risk factors generated by asymptomatic B. canis infections and the efficiency of chemoprophylaxis measures taken by sled dog owners.