Conjunctival aerobic bacterial flora in healthy Silesian foals and adult horses in Poland.

BACKGROUND: Commensal bacterial and fungal flora of the conjunctival sac has been described in horses and other animals. The identification of commensal flora of the conjunctival sac may aid in the diagnosis of ocular inflammatory diseases, such as conjunctivitis or more severe ulcerative keratitis, common in horses. Moreover, damage of ocular protective barriers may lead to an opportunistic infection. The study was carried out in Silesian horses kept at a single breeding center in South-western Poland, in order to limit any breed-dependant and climate-dependant variables affecting the results. Following an ophthalmic examination that revealed no abnormalities, sterile swabs were collected from conjunctival sac in 26 adult horses and 11 foals. The obtained swabs were subjected to bacterial culture testing. In case of Staphylococcus spp. isolation, susceptibility to methicillin was evaluated. RESULTS: Forty- three bacterial isolates, representing eleven genera of bacteria were cultured from 30 (81%) horses. Gram-positive bacteria were the dominant isolates (72%) (p < 0.001). The most commonly isolated Gram-positive bacteria were Staphylococcus spp., while Moraxella spp. were the most frequently isolated Gram-negative bacteria. There was no significant influence of sex and age on the frequency and type of microbial isolates. CONCLUSIONS: Commensal flora is present in the conjunctival sac of healthy horses in Poland. Age does not affect the abundance and type of microbial isolates.

Chlamydia prevalence in Polish pig herds.

Chlamydiae are frequently encountered intracellular Gram-negative bacteria. In pigs, these bacteria in combination with other pathogens contribute to the induction of a multi-aetiological syndrome. One of the major characteristics of Chlamydia spp. is their ability to cause prolonged, often subclinical infections. While the economic consequences of Chlamydia spp. infections in pig farms are not fully established, we know that reproductive disorders and other syndromes correlated with Chlamydia infection can lead to financial loss as a result of a reduction in pork production. Additionally, Chlamydia spp. presents a potential zoonotic hazard, therefore determining the prevalence of Chlamydia in pig populations is critical. In the present study 97 pig herds from Poland were involved. To determine the prevalence of Chlamydia PCR and CFT tests were used. In total 797 vaginal samples, 797 conjunctival samples, and 235 serum samples were collected and tested. The study took place from 2011 to 2014. We found Chlamydia spp. present in 71·2% of all tested farms. The percentage of animals testing positive on any given farm varied from 20% to 100%.

Characteristics of the bacterial flora in the conjunctival sac of cats from Poland.

OBJECTIVES: To assess the bacterial flora of the conjunctival sac in clinically healthy cats and cats with signs of conjunctivitis. METHODS: A total of 324 conjunctival swabs were examined between 2011 and 2012 taken from 60 animals, 30 of which were clinically healthy and 30 with signs of chronic conjunctivitis. The samples were taken three times at 4-week intervals from the clinically healthy cats. The samples from the cats with conjunctivitis were taken before and 4 weeks after cessation of successful therapy. Swabs from both the right and left eye of each cat were subjected to microbiological examination and polymerase chain reaction for the presence of DNA of Chlamydophila felis and Mycoplasma felis. RESULTS: There was no qualitative difference in the eye microflora between the clinically healthy animals and those with signs of conjunctivitis. Staphylococcus epidermidis (21 · 9%) was the most common microorganism isolated and it was more commonly detected in swabs from cats with conjunctivitis (P < 0 · 0001) as was Staphylococcus aureus (P = 0 · 07). The presence of C. felis was significantly correlated with (P < 0 · 0001) signs of conjunctivitis and was detected in 25% of swabs collected from both conjunctival sacs. No DNA of M. felis was detected in any swab. None of the animals had sterile conjunctival sacs in all consecutive bacteriological tests. CLINICAL SIGNIFICANCE: The conjunctival sac in cats was sterile in over 50% of the clinically healthy cats and 25% of the cats with conjunctivitis. The sterility did not persist for longer than 4 weeks. Positive bacterial cultures occur in cats with and without clinical signs of conjunctivitis.

Insight into behavior of epithelial cells of the feline conjunctiva in chronic conjunctivitis as a possible limitation in detection of Chlamydophila spp.

The aims of this work was documentation of the reactivity of feline conjunctival epithelial cells in chronic conjunctivitis and the investigation of a possible correlation of histological findings in conjunctiva with a limitation in detection of the pathogen. In this observational study, conjunctival swab samples collected from six cats suffering from chronic conjunctivitis were monitored for Chlamydophila spp. infection for one month, every ten days. Chlamydophilosis was diagnosed by conventional PCR, and confirmed by sequencing analysis. A lack of coherence with results in subsequent studies using PCR did not allow an accurate diagnosis. Additional bioptat samples of conjunctiva were collected for diagnostic purposes and stained in haematoxylin and eosin following the Giemsa method for light microscopic analysis. Additionally the samples were incubated for 15 min with IMAGEN Chlamydia conjugate (IMAGEN Chlamydia reagent kit, Dako, UK), allowing immunofluorescence detection of Chlamydophila spp. Within the epithelium an increased number of goblet cells, as well as general enlargement of the epithelium and a reduced number of normal epithelial cells, was observed. Only in areas of low epithelium could structures similar to the elementary bodies of Chlamydophila spp. be distinguished. The presented data document a possible limitation in molecular evidence for chlamydophila infection in some naturally infected cats, taking into account histological conditions in conjunctiva at the same time.

Rapid detection of Chlamydia/Chlamydophila group in samples collected from swine herds with and without reproductive disorders.

The study was carried out in seven reproductive herds of pigs. In three of them reproductive disorders were observed. Three herds consisted of 10-50 and four consisted of 120-500 adult sows and they were called small and medium, respectively. Fifty-seven adult sows were randomly selected from herds. Serum samples were tested using the complement fixation test and swabs from both eyes and from the vaginal vestibule were examined using real-time PCR. All serum samples were negative. Infected sows were present in each of the study herds. In total, there were 28 positive samples (53%, 28/48) in real-time PCR in sows with reproductive disorders and 35 (53%, 35/66) in sows selected from herds without problems in reproduction. One isolate proved to be Chlamydophila pecorum, whereas all the remaining were Chamydia suis.

Gastrointestinal nematodes in grazing dairy cattle from small and medium-sized farms in southern Poland.

This study aimed to estimate the prevalence of gastrointestinal nematodes and the intensity of infection in grazing dairy cattle from small and medium-sized farms in southern Poland. The level of antibodies against Ostertagia ostertagi in the bulk tank milk (BTM) from the animals was also assessed. Rectal fecal samples collected from 361 cows on 20 farms were examined using Willis-Schlaaf flotation and the McMaster method. BTM samples were tested for the presence of O. ostertagi antibodies using ELISA. Multiplex PCR was used to identify the third-stage larvae (L3) of gastrointestinal nematodes derived from the culture of pooled fecal samples from sampled farms. Gastrointestinal nematode eggs were found in the samples from 18 of the 20 herds with a prevalence range from 20.4 to 94.5%. The average number of eggs excreted in the feces of the herds was 200 eggs per gram (EPG). Antibodies to O. ostertagi were found in 20 of the examined herds (100%), of which 6 had optical density ratios (ODR) greater than 0.5. PCR results showed the presence of three nematode species: Ostertagia ostertagi, Cooperia oncophora and Oesophagostomum radiatum.

Reduction of prevalence of persistent BVDV infection in cattle herds by long-term vaccination program (preliminary clinical study).

Effectiveness of long-term anti-BVDV vaccination program in reducing prevalence of persistent BVDV infection in cattle herds was evaluated in seven years observational study (2005-2011). Among three seropositive dairy cattle herds (within herd seroprevalence 100%, confirmed by ELISA Herd Check BVDV Ab, IDEXX, Sweden) vaccination program based on inactivated vaccine (cytopathic strain 5960) was commenced in 2007 in two herds and continued till 2010. In the years 2007-2011 all calves aged 2-12 weeks in all three herds were tested yearly with RT-PCR in order to detect persistently infected individuals. For the entire study period true prevalence of BVDV persistent infection was significantly lower in vaccinated than in non-vaccinated herd. This may imply the role of long-term vaccination program in reducing prevalence of persistent BVDV infection in cattle herds.

Seroprevalence of BHV-1 (bovine herpesvirus type 1) among non-vaccinated dairy cattle herds with respiratory disorders.

The objective of this study was to estimate a herd-level seroprevalence of bovine herpesvirus type 1 (BHV-1) in herds with clinical symptoms of the respiratory tract. Eighty-three herds with suspected BHV-1 infection were selected and divided into two categories with respect to their size: small (n = 27) and large herds (n = 56). Samples were collected from calves, heifers and cows older than 24 months. Seroprevalence was determined using the gB ELISA test. The herd level seroprevalence was estimated as 53% (44/83) in the tested herds, 11.1% (3/27) in the small herds and 73.2% (41/56) in the large herds. Our study suggests that the current biosecurity measures still warrant improvement.

A survey of anti-Ostertagia ostertagii antibody levels in bulk tank milk samples (BTM) in dairy herds in Lower Silesia Region (Poland).

This work presents serological evidence of cattle ostertagiosis in the Lower Silesia Region (Poland), based on the measurement of antibodies in bulk tank milk (BTM) samples. It represents the first evidence of this parasite examined with the use of the ELISA test and milk samples in Poland. The prevalence of Ostertagia ostertagii antibodies was determined in BTM from 32 dairy cattle herds. Antibodies to O. ostertagii were demonstrated in all herds. The optical density ratio (ODR) varied from -0.088 to 1.024. The mean ODR value in the examined region was 0.53.

Real-time PCR detection of Mycoplasma felis in domestic cats suffering from chronic conjunctivitis (Poland).

Real-time PCR directed to intergenic spacer (IGS) noncoding region between 16S and 23S rRNA genes was used for species specific detection of Mycoplasma felis in conjunctival scrapings. Samples were collected from 57 cats suffering from chronic conjunctivitis in 2008-2010 (Wroclaw, Poland). Samples from 36 cats (63.2%) were shown to be positive for Mycoplasma felis. Our research gives a first insight in the occurrence of Mycoplasma felis among domestic cats in Poland suggesting that this pathogen may constitute an underestimated cause of chronic conjunctivitis.

Feline herpesvirus 1 and Chlamydophila felis prevalence in cats with chronic conjunctivitis.

The prevalence of Chlamydophila felis and Feline Herpesvirus type 1 was investigated in 30 cats with chronic conjunctivitis, with use of conjunctival swabs and conventional polymerase chain reaction (PCR). In cats with chronic conjunctivitis the DNA of C. felis and FHV-1 was detected in 2 of 30 cats (6.7%) and in 10 of 30 animals (33.3%), respectively. One case of FHV-1 DNA, and none of C. felis was found in control group. There was no case of co-infection with both pathogens.