Impact of the Application of Gaseous Ozone on Selected Pathogens Found in Animal Shelters and Other Facilities.

Correctly selecting disinfection procedures is crucial in facilities housing a high number of animals as it directly affects their health. The aim of this study was to verify the virucidal effect of gaseous ozone delivered by commercially available generators under controlled experimental conditions on a selection of viral pathogens (feline coronavirus, canine coronavirus, feline calicivirus, feline parvovirus) commonly found in shelters and other facilities. Two ozone generators with outputs of 3.5 g/h and 20 g/h were used to produce ozone. Virus viability after the application of ozone was evaluated by examining for typical pathogen-specific cytopathic effects on the CRFK (Crandell-Rees Feline Kidney) cell line post-incubation. No cytopathic effect was observed in feline coronavirus after the 2-h application of ozone; in canine coronavirus, the absence of a cytopathic effect was observed after the 4-h application of ozone. The absence of a cytopathic effect in feline calicivirus was observed after the 6-h application of ozone; the viability of feline parvovirus was not impaired even by the 6-h application of ozone. The results of the study confirm lower resistance to the application of gaseous ozone in enveloped viruses.

Shedding persistency and intensity patterns of feline coronavirus (FCoV) in feces of cats living in breeding catteries in the Czech Republic.

Infection with feline coronavirus (FCoV) is a major problem in multiple-cat households, where many cats are kept together in a small space such as catteries and shelters. Sixty cats from 19 breeding catteries included in the study were evaluated for their shedding persistency and intensity patterns using qPCR identification of FCoV in feces. Cats were identified based on shedding persistency as non-shedders (NS) if all four samples negative, intermittent shedders (IS) when at least one positive and one negative sampling followed by another positive sampling, persistent shedders (PS) if all four samples positive and shedders with unclear status (US) if the shedding patterns could not be determined based on only 4 samples. There were 11 NS (18%), 15 IS (25%) and 15 PS (25%) and in 19/60 cats (32%), the shedding patterns could not be determined based only on four samplings. The intensity of shedding was evaluated based on the total number of FCoV particles shed during the 12 months of the study. There were 11 non-shedders (18%), 2 very low intensity shedders (3%), 9 low intensity shedders (15%), 25 medium intensity shedders (42%) and 13 high intensity shedders (22%). Intermittent shedders were shedding significantly lower FCoV particles compared to the persistent shedders (p = 0.0082). Permanent shedders represent the most important source of FCoV infection in multi-cat households and identifying permanent shedders in is the key to minimize the viral load in the environment to control FCoV in a shelters and breeding catteries.

Direct Detection of Feline Coronavirus by Three Rapid Antigen Immunochromatographic Tests and by Real-Time PCR in Cat Shelters.

The aim of this study was the direct detection of feline coronavirus by real-time PCR and by three different rapid immunochromatographic (RIM) tests detecting antigens in faecal samples of shelter cats. Based on sensitivity and specificity calculated for each of the RIM tests, the utility of RIM tests was compared. Seventy faecal samples originating from shelter cats housed in quarantine were examined. Out of 70 samples analyzed by real-time PCR, 44 (62.9%) were positive. Significantly more cats (p < 0.05) tested positive than negative. Neither age nor sex of the cats played a significant role (p > 0.05) in the shedding status of the virus. The sensitivity of the RIM tests was found to be at low (<35%; RIM tests A and C) to satisfactory level (>50%, RIM test B). The number of virus particles determined by real-time RT-PCR analysis did not significantly correlate with the results detected by any of the RIM tests (p > 0.05). The results of this study indicate that the use of rapid antigen RIM tests in routine screening of FCoV shedding status in shelter cats is limited due to the occurrence of a high number of false negative results.

Candidate Gene Markers Associated with Fecal Shedding of the Feline Enteric Coronavirus (FECV).

The Feline coronavirus (FCoV) can cause a fatal disease, the Feline Infectious Peritonitis. Persistent shedders represent the most important source of infection. The role of the host in FCoV fecal shedding is unknown. The objective of this study was to develop gene markers and to test their associations with FCoV shedding patterns. Fecal samples were taken from 57 cats of 12 breeds on the day 0 and after 2, 4 and 12 months. Variation from persistent and/or high-intensity shedding to no shedding was observed. Thirteen immunity-related genes were selected as functional and positional/functional candidates. Positional candidates were selected in a candidate region detected by a GWAS analysis. Tens to hundreds of single nucleotide polymorphisms (SNPs) per gene were identified using next generation sequencing. Associations with different phenotypes were assessed by chi-square and Fisher's exact tests. SNPs of one functional and one positional candidate (NCR1 and SLX4IP, respectively) and haplotypes of four genes (SNX5, NCR2, SLX4IP, NCR1) were associated with FCoV shedding at pcorected < 0.01. Highly significant associations were observed for extreme phenotypes (persistent/high-intensity shedders and non-shedders) suggesting that there are two major phenotypes associated with different genotypes, highly susceptible cats permanently shedding high amounts of viral particles and resistant non-shedders.

Expression and serological reactivity of hemorrhagic enteritis virus hexon protein.

The aim of this work was to express the recombinant hexon protein of the hemorrhagic enteritis virus, to establish the diagnostic value of this protein for serological detection of antibodies in turkey serum samples and to assess seroprevalence of the infection in the Czech Republic. The N' terminal part of the hexon protein was expressed in a bacterial expression system and used as an antigen in an ELISA test for the detection of hemorrhagic enteritis virus specific antibodies in turkey sera. Validation of the test was performed by comparison with a commercially available ELISA test. Serological reactivity was assessed on a panel of 126 turkey sera by a newly developed ELISA test. Serum samples were taken from turkey farms with the history of hemorrhagic enteritis virus infection, from farms with animals free of infection, and from turkey farms following vaccination. Both ELISA kits gave identical results (100 %) with the tested sera. ELISA based on the recombinant hexon protein thus proved useful and cheaper for detection of antibodies in turkey flocks infected with the hemorrhagic enteritis virus.

[Human intestinal spirochetosis]. / Intestinální spirochetóza cloveka.

A characteristic feature of human intestinal spirochetosis (IS) is the colonization of the mucosa of the large intestine with intestinal spirochetes of the genus Brachyspira. The joining of the brachyspirae with the apical cellular membrane of enterocytes resembles in histological slides a false brush border of the intestinal mucosa. Various symptoms related to the involvement of the large gut were found with invasive IS. From the cultures of these cases were isolated Brachyspira aalborgii and B. pilosicoli. The frequency of the incidence of brachyspirae depended on the socio-economic living conditions of people. Colonization of the mucosa of the large gut was found more often in human populations in the developing countries; it was fairly rare in countries with high hygienic standards. An exception were men of homosexual orienation and patients presenting with a HIV infection. Isolation of brachyspirae from the faeces and biopsy of the mucosa of the large gut are fairly demanding jobs, especially with B. aalborgii. Most documented IS cases of this aetiology were diagnosed using immunohistochemical methods and amplification of the genus-specific region of the gene 16S rRNA. Isolation of B. pilosicoli tends to be simpler, it requires anaerobic incubation on selective blood agars for a period of 3-6 days at 37 degrees C. When manual haemoculture systems were used, patients in a critical state presented a translocation of brachyspirae into blood circulation, while automatic systems don't necessarily diagnose spirochetaemia. In the management of described cases of invasive IS particularly successful proved metronidazole and beta-lactam antibiotics. In isolated B. pilosicoli, in vitro tests confirmed sensitivity to metronidazole, ceftriaxone, meropenem, tetracycline, moxifloxacine and chloramphenicol. A varying frequency of resistance was found with clindamycin and amoxicillin, which how ever was efficacious in combination with clavulanic acid.

Decreased susceptibility to tiamulin and valnemulin among Czech isolates of Brachyspira hyodysenteriae.

The agar dilution method was used to investigate the sensitivity to pleuromutilins of 100 isolates of Brachyspira hyodysenteriae isolated from 63 pig farms between 1997 and 2001. In the period under investigation, MICs to both tiamulin and valnemulin increased, with differences between the periods 1997-98 and 1999-2001 being statistically significant (P < 0.001 for tiamulin and P < 0.0001 for valnemulin). Between 1997 and 2001, the MIC50 and MIC90 of tiamulin increased from 0.062 and 0.25 microg ml, respectively, to 1.0 and 4.0 microg ml. Valnemulin MIC50 and MIC90 were < or = 0.031 microg ml in 1997 and by 2001 were respectively, 2.0 and 8.0 microg ml. The increase in MICs of tiamulin and valnemulin demonstrated in this study reflect the intensity of pleuromutilin use in the treatment of swine dysentery in the Czech Republic.