Prevalence and Diversity of Haemotropic Mycoplasma Species in Cats and Their Ectoparasites (Fleas and Ticks).

Mycoplasma spp. pathogens frequently cause chronic and acute diseases in cats. The aim of the present study was to investigate the presence and genetic diversity of Mycoplasma spp. in cats and their ectoparasites using PCR and sequence analysis of the 16S rRNA gene. Blood samples were collected from 541 domestic and stray cats in Lithuania. Ectoparasites (153 fleas and 321 ticks) were collected from owned domestic cats that live both outdoors and indoors. Mycoplasma spp. were detected in 7.2% of cat blood samples and 4.4% of Ctenocephalides felis fleas. The sequence analysis revealed the presence of Mycoplasma haemofelis in 1.1% of cats and 'Candidatus Mycoplasma haematominutum' in 4.8% of cats. Ct. felis fleas harboured M. haemofelis. To the best of the authors' knowledge, this is the first report on the prevalence and molecular characterisation of Mycoplasma bacteria in cats in Lithuania and cat fleas in the Baltic States.

An ELISA for the early diagnosis of acute canine babesiosis detecting circulating antigen of large Babesia spp.

Babesia canis is the predominant Babesia species in dogs in Europe and is responsible for a severe and fatal disease. An increase in global pet tourism and a widening of the geographic distribution of the tick vector has led to the emergence of infections in areas where previously only imported cases have been reported. Due to the potential for rapid and serious disease progression, direct parasite detection by stained blood smears and light microscopy or DNA-based methods have traditionally been used for the diagnosis of acute infections. This study describes the production of a murine monoclonal antibody ('mAb BcFIII 7/1/2') that reacts to a 65kDa corpuscular epitope present in B. canis-infected erythrocytes and can be used in an ELISA to detect circulating Babesia antigen during acute infections. The sensitivity of the ELISA was 100% (95%CI: 84.5-100) as determined using blood lysate samples from 27 dogs with acute B. canis infections. Sensitivity was reduced to 53.8% in 13 patent Babesia vogeli infections (95%CI: 26.1-79.6) based on the current test design using convalescent serum from a B. canis-infected dog. The specificity was determined to be 86.4% (95%CI: 64-96.4) using 22 samples from healthy canine blood donors. In the course of acute B. canis infections, the ELISA showed a positive result at the same time as a positive PCR result was recorded. This was 24-48h before parasites could be detected by light microscopy. Convalescent samples collected from 6 B. canis-infected dogs at least 14days post treatment resulted in negative ELISA reactions. The hyper-acute to acute phase of a B. canis infection represents an emergency situation with high mortality. To increase the chances of survival, a fast and accurate diagnosis and immediate treatment is required. The current study demonstrates the opportunity of an early and specific detection of acute infections by an AgELISA that is potentially translatable to a rapid diagnostic test design.

DETERMINATION OF ANTIMICROBIAL ACTIVITY OF CHLORHEXIDINE GEL.

Many researchers have advocated in recent times that antiseptic use in healing wounds should be discouraged. Antiseptics have been found to retard healing of wounds. Poloxamer 407 shows thermoreversible properties, which are of the utmost interest in optimizing drug formulation (fluid state at room temperature facilitating administration and gel state above sol-gel transition temperature, at body temperature, promoting prolonged release of pharmacological agents). Chlorhexidine, a commonly used antiseptic, is known to be less toxic on granulation cells. Acting as an antiseptic, it is an effective bactericidal agent against the most categories of microbes, including bacteria, yeast, and viruses. Objective of this study was to evaluate antimicrobial activ- ity of chlorhexidine containing poloxamer gel to Gram-positive and Gram-negative bacteria in vitro. Chlorhexidine gels and chlorhexidine aqueous solutions have different antibacterial activity to S. amis, E.faecalis, E. coli and P. aemginosa strains in vitro. It depends on concentration and dosage form of antiseptic. Study results confirmed that antimicrobial activity of gel depends on active ingredient concentration in antiseptic. The best inhibition effect for both of reference and wild-type bacteria was obtained for 1% chlorhexidine gel. Summarizing the results and assessing the characteristics of the gel ingredients, it can be suggested using chlorhexidine gels in veterinary medicine.

Detection and molecular characterization of canine babesiosis causative agent Babesia canis in the naturally infected dog in Lithuania.

Canine babesiosis caused by Babesia canis is an emerging infectious disease in Europe. Although previously uncommon, canine babesiosis has become quite frequent in Lithuania during the past decade. In the last few years an increasing number of cases with a wide variety of clinical signs have been recorded throughout the country. In Lithuania the identification of the disease agent in veterinarian clinics is based on a microscopic analysis of size and morphology. To date, no data on the genetic characterization of Babesia species in dogs have been documented for Lithuania. A total of 123 blood samples from dogs showing clinical signs of babesiosis on the basis of veterinary examination were tested for the presence of babesial parasites. Babesia isolated from dogs were detected and characterized by nested-PCR and sequence analysis of a fragment of the 18S rRNA gene. Babesia parasites were detected in blood smears of 94 dogs (76.4%). The molecular analysis revealed the presence of B. canis in 108 dogs (87.8%). Two genotypes of B. canis were distinguished on the basis on two nucleotide (GA → AG) substitutions observed in 18S rRNA gene sequences. The results of the present study provide knowledge of the distribution of B. canis genotypes in dogs in Lithuania, and show the necessity to use a molecular analysis for an accurate diagnosis of canine babesiosis.

Evaluation of peculiarities of the acetylcholinesterase-positive nerve plexus and its length in the cornea.

UNLABELLED: The objective of this study was to evaluate age-related peculiarities of acetylcholinesterase-positive nerve plexus and its length in the dog cornea. MATERIAL AND METHODS: Nine mixed breed dogs (male and female) of 1.8-20 kg weight were used. They were divided into three age groups: young (until 1 year), adult (1-7 years) and old (8 and more years). Acetylcholinesterase-positive nerve plexus was identified using acetylcholinesterase method of M. J. Karnowsky and L. A. Roots (1964) modified by D. H. Pauza et al. (1996). The length of nerve bundles was measured by mm in 1 mm(2) of cornea. RESULTS AND CONCLUSIONS: Branching thick, medium and thin nerve bundles form acetylcholinesterase-positive nerve plexus in dog cornea. The average of the length of nerve bundles between the left and the right corneas was similar in different age groups (p>0.05). The length of acetylcholinesterase-positive nerve bundles in 1 mm(2) of the cornea in the group of adult dogs (10.32+/-0.11 mm) was higher than in the groups of young (9.42+/-0.02 mm) and old dogs (7.75+/-0.14 mm) (p<0.001).