Wojciech Zygner

Prevalence of Babesia canis, Borrelia afzelii, and Anaplasma phagocytophilum infection in hard ticks removed from dogs in Warsaw (central Poland).

The purposes of this study were to specify the occurrence and prevalence of Babesia canis, Borrelia burgdorferi sensu lato, and Anaplasma phagocytophilum in ticks removed from dogs in Warsaw, and to determine the Borrelia species occurring in Ixodes ricinus ticks. Among 590 collected ticks, 209 were identified as I. ricinus, and 381 as Dermacentor reticulatus. DNA of B. canis was detected in 11% of D. reticulatus ticks. We found that 6.2% of I. ricinus ticks harbored B. burgdorferi s.l. specific DNA and 2.9% harbored A. phagocytophilum DNA. In these samples sequencing of the detected Borrelia amplicon confirmed infection with Borrelia afzelii genospecies. New sequences were submitted to the GenBank database (accession no. EU152128, EU152127, EU152126). This work is the first detection of B. afzelii and A. phagocytophilum in ticks from Warsaw, and the first survey for the prevalence of B. canis, B. afzelii, and A. phagocytophilum in ticks in central Poland.

Detection of the DNA of Borrelia afzelii, Anaplasma phagocytophilum and Babesia canis in blood samples from dogs in Warsaw

Each month, from March 2003 to February 2004, 34 blood samples from dogs were randomly selected from the blood samples delivered to two veterinary laboratories in Warsaw and tested for the DNA of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Babesia canis and Hepatozoon canis. Borrelia DNA was detected in seven of the 408 dogs, A phagocytophilum DNA was found in two, and B canis DNA was found in 48 (11.8 per cent). The DNA of H canis was not found in any of the blood samples. Sequencing of the seven Borrelia amplicons showed that only the genospecies Borrelia afzelii was present, the first time it has been detected in dogs in Poland.

Increased concentration of serum TNF alpha and its correlations with arterial blood pressure and indices of renal damage in dogs infected with Babesia canis

Canine babesiosis is a tick-borne disease caused by parasites of the genus Babesia. Tumour necrosis factor alpha (TNF-α) is a cytokine that plays a role in the pathogenesis of canine babesiosis. In this study, the authors determined the concentration of serum TNF-α in 11 dogs infected with Babesia canis and calculated Spearman’s rank correlations between the concentration of TNF-α and blood pressure, and between TNF-α and indices of renal damage such as: fractional excretion of sodium (FE(Na+)), urinary creatinine to serum creatinine ratio (UCr/SCr), renal failure index (RFI), urine specific gravity (USG) and urinary protein to urinary creatinine ratio (UPC). The results demonstrated statistically significant strong negative correlations between TNF-α and systolic arterial pressure (r = −0.7246), diastolic arterial pressure (r = −0.6642) and mean arterial pressure (r = −0.7151). Serum TNF-α concentration was also statistically significantly correlated with FE(Na+) (r = 0.7056), UCr/SCr (r = −0.8199), USG (r = −0.8075) and duration of the disease (r = 0.6767). The results of this study show there is an increase of serum TNF-α concentration during canine babesiosis, and the increased TNF-α concentration has an influence on the development of hypotension and renal failure in canine babesiosis. This probably results from the fact that TNF-α is involved in the production of nitric oxide and induction of vasodilation and hypotension, which may cause renal ischaemia and hypoxia, and finally acute tubular necrosis and renal failure.

Blocking Fasciola hepatica's energy metabolism - a pilot study of vaccine potential of a novel gene - phosphoglycerate kinase.

Fasciola hepatica infections cause huge economic losses to livestock production and are a serious problem in human and veterinary medicine. The main difficulty in the control of infections is progressing drug resistance. Moreover, pharmacological therapy is expensive and harmful for the environment. The best way of prophylaxis against infections seems to be vaccination. A new generation of vaccines could be the best possible way of controlling fasciolosis. This paper is focused on first vaccination trials based on a new vaccine candidate antigen, F. hepatica phosphoglycerate kinase (FhPGK) performed on a rat model. We obtained protection levels ranging from 0% to 69% depending on the way of delivery and form of vaccine.

Molecular cloning and characterisation of in vitro immune response against astacin-like metalloprotease Ace-MTP-2 from Ancylostoma ceylanicum.

Ancylostoma ceylanicum belongs to the group of parasites commonly known as hookworms, blood-sucking nematodes which infect around 576 million people and hundreds of millions of animals. The interactions between these parasites and host immune systems are complicated and yet to be determined. Hookworm infections are usually long lasting and recurrent, due in part to their ability to synthesize macromolecules capable of modulating the host immune response. The interaction of parasite proteins with host immune systems has been proven, but so far there is no data describing the influence of astacin-like metalloproteases (expressed among different parasitic nematodes) on the human immune system. The cDNA encoding A. ceylanicum metalloprotease 2 (Ace-mtp-2) was cloned using RACE-PCR. Computational analysis was used to examine the immunogenicity and recombinant Ace-MTP-2 was used to investigate its influence on human THP-1 monocytes and macrophages. The Ace-mtp-2 gene encodes an astascin-like metalloprotease, with a theoretical molecular mass of 26.7 kDa. The protease has a putative signal peptide, 11 potential phosphorylation sites, and two disulfide bridges revealed by computational analysis. Maximal expression of Ace-mtp-2 by A. ceylanicum occurs in the adult stage of the parasite, and Western blot indicates the secretory nature of the protease. This suggests the protease is working at the host-parasite interface and would likely be exposed to the hosts immune response. Recombinant protein were expressed in Escherichia coli and Pichia pastoris. Recombinant Ace-MTP-2 amplified the in vitro release of TNFα and induced release of IFNγ by lipopolysaccharide activated THP-1 macrophages. The presence of Ace-MTP-2 in secretory products of the adult parasite and the induction of IFNγ release may suggest an important role for Ace-MTP-2 in host-parasite interactions since IFNγ is suggested to be responsible for the protective immune response against adult hookworms.

Evaluation of the immune response of male and female rats vaccinated with cDNA encoding a cysteine proteinase of Fasciola hepatica (FhPcW1)

Not only do males and females of many species vary in their responses to certain parasitic infections, but also to treatments such as vaccines. However, there are very few studies investigating differences among sexes following vaccination and infection. Here we demonstrate that female Sprague-Dawley rats vaccinated with cDNA encoding a recently discovered cysteine proteinase of Fasciola hepatica (FhPcW1) develop considerably lower liver fluke burdens after F. hepatica infection than their male counterparts. This is accompanied by differences in the course of their immune responses which involve different eosinophil and monocyte responses throughout the study as well as humoral responses. It is evident that host gender influences the outcome of parasitic infections after vaccination and research on both sexes should be considered when developing new treatments against parasites.

Strong monovalent electrolyte imbalances in serum of dogs infected with Babesia canis

Canine babesiosis is a systemic tick-borne protozoan disease caused by infection with parasites of the genus Babesia. Acid-base disorders and ion imbalances have been described in dogs infected with Babesia rossi in South Africa. In this paper, the authors describe changes to monovalent ion concentrations and calculated parameters of monovalent ions in 70 dogs naturally infected with B. canis, a species occurring in Europe. Hyponatraemia, hypokalaemia, hyperchloraemia, decrease of chloride gap, strong ion gap, difference between sodium and chloride concentrations, and an increase of chloride-to-sodium and sodium-to-potassium ratios were the most prevalent changes. Hyponatraemia, hypokalaemia and hyperchloraemia were detected less frequently than in dogs infected with B. rossi, but the severity of these changes were similar. Comparison of monovalent ion concentrations in azotaemic and non-azotaemic, and anaemic and non-anaemic dogs infected with B. canis showed that azotaemic dogs had significantly lower sodium concentrations. The results of this study indicate a possible development of hyperchloraemic acidosis and the probable contribution of aldosterone in the development of hypokalaemia. However, further study on blood gas, aldosterone, and antidiuretic hormone in dogs infected with B. canis is needed.

Low T3 syndrome in canine babesiosis associated with increased serum IL-6 concentration and azotaemia

Low triiodothyronine (T3) syndrome, also named euthyroid sick syndrome or non-thyroidal illness syndrome, has been recognized in canine babesiosis caused by Babesia rossi, where it manifested by lowering of the serum thyrotropin (TSH), total thyroxin (TT4) and free thyroxin (FT4) concentrations. This syndrome has also been observed in critical diseases in humans and animals, and the severity of the disease is considered an important factor in lowering of thyroid hormone concentrations. Interleukin-6 (IL-6) plays a role in the development of low T3 syndrome by causing a decrease in deiodinases 1 and 2 activity and increased activity of deiodinase 3, enzymes involved in the conversion of thyroxin (T4) to T3. The purpose of this study was to compare the concentrations of serum thyroid hormones and TSH between healthy dogs and dogs with babesiosis, and to determine correlations between serum IL-6 concentration and serum total T3 (TT3), TT4, FT4, and TSH concentrations, and the level of azotaemia in dogs with babesiosis. The concentrations of IL-6, TT3, TT4, FT4, TSH, urea and creatinine were determined in 13 dogs with canine babesiosis caused by Babesia canis and in 10 healthy dogs. The results of this study showed decreases in TT3, TT4, FT4, and TSH and increases in IL-6, urea and creatinine concentrations in affected dogs in comparison to healthy dogs. The concentration of IL-6 was negatively correlated with TT3 and TSH concentrations and the TT3 concentration was negatively correlated with serum urea and creatinine concentrations. This study showed low T3 syndrome in canine babesiosis, which was confirmed by the determination of the T3 concentration, and demonstrates that in canine babesiosis the T3 concentration is associated with IL-6 concentration.

Immune responses in rats and sheep induced by a DNA vaccine containing the phosphoglycerate kinase gene of Fasciola hepatica and liver fluke infection.

Immune responses of rats and sheep following vaccination with cDNA encoding phosphoglycerate kinase of Fasciola hepatica (cDNA-FhPGK/pCMV) and F. hepatica infection were investigated in the present study. cDNA-FhPGK/pCMV vaccinated female Sprague-Dawley rats were better protected by vaccination than their male counterparts–48% reduction in fluke burden for females and no protection for males when compared with appropriate infection control groups. Moreover, male rats developed marked leukocytosis during the study with higher neutrophil, eosinophil and monocyte responses than females. Additionally, dynamics of eosinophil and monocyte responses varied between sexes. Increased titres of anti-FhPGK IgG1 and IgG2a correlated with the protective effect of vaccination that was observed among female rats. In the case of male sheep, no differences in worm burdens and in the course of the immune response were observed following vaccination. Titres of specific antibodies detected were low, and cellular responses were not significant. Apparently, sheep immune responses induced by cDNA-FhPGK/pCMV vaccination are not effective at controlling F. hepatica infection. Poor immunogenicity of DNA vaccines in large animals is still a major obstacle of this technology that has to be overcome.

Prevalence of feline hyperthyroidism in mature cats in urban population in Warsaw

Abstract The purpose of this study was to assess the prevalence of feline hyperthyroidism in a cat population in Warsaw, considering risk factors. The study was conducted between June 2007 and July 2011. Seven-year-old and older cats were examined. Diagnosis of feline hyperthyroidism was based on the results of clinical examination, data from clinical history, and serum concentrations of thyroid hormones. Hyperthyroidism was diagnosed in 20.14% of 417 cats (95% confidence interval (CI): 16.28%-24.01%). Statistically significant risk factors were age (odds ratio (OR) = 1.17; 95% CI: 1.08-1.27), feeding with a commercial wet feed (OR = 6.74, 95% CI: 2.03-22.37), and an indoor lifestyle (OR = 2.25, 95% CI: 1.04-4.84). There were no effects of breed, gender, castration, or the frequency of deworming on the occurrence of hyperthyroidism. Feline hyperthyroidism in Warsaw is a prevalent disease. This probably results from chronic exposure to dietary and environmental factors.