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1.
OBJECTIVE: To detect and characterize the full range of chlamydial infections in cats with ocular disease by use of polymerase chain reaction (PCR) assays, cytologic examination, immunohistochemical analysis, and evaluation of clinical information including status for feline herpesvirus-1 (FeHV-1). SAMPLE POPULATION: DNA extracted from 226 conjunctival samples obtained from cats with clinically diagnosed keratitis or conjunctivitis and 30 conjunctival samples from healthy cats. PROCEDURE: PCR assays for the 16S rRNA gene specific for the order Chlamydiales and a new Chlamydophila felis (formerly Chlamydia psittaci) species-specific 23S rRNA gene were performed. Seventy-four conjunctival samples were prepared with Romanowsky-type stain, grouped on the basis of inflammatory pattern, and screened for chlamydial inclusions by use of immunohistochemical analysis. Clinical information and FeHV-1 status were recorded. RESULTS: 26 (12%) specimens had positive results for the only known feline chlamydial pathogen, C felis. Surprisingly, an additional 88 (39%) were positive for non-C felis chlamydial DNA. Identification of non-C felis chlamydial DNA by direct sequencing revealed 16S rRNA gene sequences that were 99% homologous to the sequence for Neochlamydia hartmannellae, an amebic endosymbiont. Chlamydial prevalence was significantly higher in cats with ocular disease. CONCLUSIONS AND CLINICAL RELEVANCE: Application of a broad-range detection method resulted in identification of a new agent associated with ocular disease in cats. Finding chlamydia-like agents such as N hartmannellae in coinfections with their obligate amebic host, Hartmannella vermiformis, raises questions about the potential role of these microorganisms in causation or exacerbation of ocular disease in cats.  相似文献   

2.
Corneal samples of cats with and without corneal diseases were screened with a pan‐Chlamydiales PCR and specific PCRs for Parachlamydia, Protochlamydia, Chlamydophila felis, Acanthamoeba and feline herpesviruses (FHV‐1). Several corneal samples tested positive for Parachlamydia and related Chlamydiales, indicating cat exposure to these intracellular bacteria.  相似文献   

3.

Background

Cytologic examination of smears prepared from ocular swabs of conjunctiva from cats with conjunctivitis permits identification of the type of inflammation and possibly specific microorganisms. Results of studies of the diagnostic utility of cytology for detection of infectious causes of feline conjunctivitis have been inconsistent.

Objectives

The objectives of this study were to describe cytologic findings in cats with conjunctivitis and to compare those findings with results of PCR analysis for feline herpesvirus (FHV‐1), Chlamydophila felis (C felis), and Mycoplasma felis (M felis).

Methods

Conjunctival smears from 88 cats with conjunctivitis and 10 healthy control cats were stained with a Romanowsky stain and evaluated for the type of inflammation and evidence of an infectious agent. PCR analysis for FHV‐1, C felis, and M felis was performed.

Results

Infectious agents identified by PCR analysis were FHV‐1 in 9 cats (10%), C felis in 8 cats (9%), and M felis in 6 cats (7%). Inclusions interpreted as chlamydial inclusions were found in all cytologic smears from cats positive for C felis by PCR analysis and in 3 PCR‐negative cats. Inclusions interpreted as Mycoplasma organisms were found in 3 of 6 cats that were PCR‐positive for M felis and in 1 PCR‐negative cat. FHV‐1 inclusion bodies were not detected on cytologic examination.

Conclusions

Cytologic examination can be diagnostic for C felis infection when many typical inclusions are present. Cytologic examination was unreliable in diagnosing M felis infection, and viral inclusions of FHV‐1 were not found in specimens stained with Romanowsky stains.  相似文献   

4.
Objective To investigate the prevalence of feline calicivirus (FCV) infection in relation to ocular surface lesions in cats with upper respiratory tract diseases (URTD). Animals studied Ninety‐nine cats with ocular surface infection and symptoms or recent history of URTD were examined at various rescue shelters and hospitals. Procedure A complete general and ophthalmic examination was performed including Schirmer tear test, slit‐lamp biomicroscopy, fluorescein and lissamine green staining. Clinical and ocular symptoms were scored and recorded. Conjunctival samples were collected using a cytobrush, and nucleic acid extraction using RT‐PCR was carried out to analyze for the presence of various infectious agents. Results RT‐PCR detected either FCV, feline herpes virus type 1 (FHV‐1), Chlamydophila felis or Mycoplasma spp. in 63/99 samples. 30/63 samples were positive for FCV, 23/63 for C. felis, 21/63 for Mycoplasma spp., and 16/63 for FHV‐1. Out of the 30 FCV‐positive samples, 11 were positive only for FCV and in 19 samples FCV was seen in combination with other agents. FCV infection was highest in animals examined at the rescue centers and in the age group of 0–2 months. Erosive conjunctivitis was an important ocular finding. Oral ulcers were detected in all FCV‐infected cats. Conclusion Results indicate that FCV is highly prevalent in cats with URTD either as a sole infectious agent or in combination with other pathogens and therefore is a potential cause for ocular surface lesions during the URTD.  相似文献   

5.
OBJECTIVES: (1) To document tear film break-up time (TFBUT) in a group of cats with conjunctivitis; (2) to determine if TFBUTs from cats with conjunctivitis vary significantly from previously established normal values for TFBUT in young cats without ocular disease; (3) to determine if a correlation exists between Schirmer tear test (STT) values and TFBUTs in cats with conjunctivitis; (4) to determine if the TFBUTs in cats with conjunctivitis are influenced by the detection of DNA from feline herpes virus-type 1 (FHV-1), Chlamydophila felis, Mycoplasma spp., and feline calicivirus. ANIMALS STUDIED: Fourteen cats between the ages of 0.8 years to 12 years with active, untreated conjunctivitis and without active keratitis or other ocular or systemic abnormalities were included in this study. Procedures Complete ophthalmic examinations, including TFBUT, were performed on all cats. Polymerase chain reaction (PCR) screening for FHV-1, Chlamydophila felis, Mycoplasma spp., and feline calicivirus was performed on conjunctival swabs from affected eyes and blood samples from all cats. RESULTS: Mean TFBUT for cats in this study was 8.9 (+/- 4.8) s in the right eye (OD) and 8.1 (+/- 4.6) s in the left eye (OS). No correlation existed between mean TFBUTs and mean STT values OD or OS. Conjunctival swabs from seven cats (n = 9 eyes) tested positive via PCR for one of the above infectious agents. Blood samples from nine cats tested positive for FHV-1. Mean TFBUTs for cats from which the DNA from FHV-1 was isolated from the blood were significantly lower than mean TFBUTs for cats from which no such DNA was isolated from the blood. CONCLUSIONS: In this study, the mean TFBUT in cats with conjunctivitis was significantly lower than previously established values for clinically healthy cats. This supports the theory that qualitative tear film deficiency, and thus tear film instability, may play a role in the pathogenesis of feline conjunctivitis. Qualitative tear film deficiency may predispose to the development of conjunctivitis or may occur secondarily to this condition.  相似文献   

6.
Nasal lavage fluid was collected from 155 tortoises, mostly Testudo spp., that were kept as companion animals and suffered from nasal discharge. Examination for chlamydial DNA by PCR assays targeting the ompA, ompB, and groESL genes, as well as the 16S rRNA signature region and the 16S-23S intergenic spacer, respectively, revealed 16 (10.3%) positive animals. Sequence analysis of PCR products indicated high homology to the family Chlamydiaceae. Phylogenetic trees constructed from partial sequences of the ompA and 16S rRNA genes showed that the present samples clustered outside the nine species of Chlamydia and Chlamydophila. Sequences of the nearest relative, Chlamydophila pecorum, were still clearly distinct from those of the positive tortoise samples. This suggests that the tortoises had been infected by Chlamydia-like agents, the taxonomic identity and pathogenic importance of which has yet to be established.  相似文献   

7.
OBJECTIVE: To use PCR assays to determine the prevalence of feline herpesvirus 1 (FHV-1), Chlamydophila felis, and Mycoplasma spp DNA in conjunctival cells collected from cats with and without conjunctivitis; to compare results of conventional and real-time fluorogenic PCR assays for amplification of FHV-1 DNA; and to determine whether copy numbers of FHV-1 DNA are correlated with conjunctivitis. ANIMALS: 55 cats with active conjunctivitis, 39 healthy cats that never had conjunctivitis, and 32 cats with a history of conjunctivitis that had been resolved for at least 3 months. PROCEDURES: Samples were obtained by rolling cotton-tipped applicators on the ventral conjunctiva of awake cats treated topically with proparacaine. The DNA was extracted from the swab specimens and assessed in PCR assays to detect DNA of FHV-1 (fluorogenic PCR assay and conventional PCR assay), Mycoplasma spp (conventional PCR assay), and C felis (conventional PCR assay). RESULTS: Overall prevalence rates of FHV-1, C felis, and Mycoplasma spp as assessed by the conventional PCR assays were 6.7%, 3.2%, and 9.6%, respectively. Percentage concordance between conventional PCR and fluorogenic PCR assays for FHV-1 was 92.5%. There were no significant differences among the 3 groups of cats for the mean copy number of FHV-1 divided by the copy number of glyceraldehyde-3-phosphate dehydrogenase. CONCLUSIONS AND CLINICAL RELEVANCE: Mycoplasma spp were the most prevalent organism detected and was associated with conjunctivitis. This study could not confirm that there are increased copy numbers of FHV-1 DNA in cats with conjunctivitis, compared with the copy numbers for cats without conjunctivitis.  相似文献   

8.
Abstract

AIMS: To obtain information and compare the prevalence of Chlamydiaceae in riverine buffalo (Bubalus bubalis) and cows (Bos taurus) in Egypt with and without clinical signs of reproductive disease.

METHODS: Vaginal swabs and blood samples were collected from animals attending Governmental Veterinary Clinics without (buffalo n=39, cows n=20) and with (buffalo n=63, cows n=53) signs of reproductive disease. Serum samples were tested for antibodies to Chlamydiaceae using complement fixation testing (CFT). Vaginal swabs were tested for Chlamydiaceae following inoculation into Vero cells and 6-day-old embryonated chicken eggs, using modified Giménez and immunoperoxidase staining, PCR analyses targeting the omp2 gene, and Restriction Fragment Length Polymorphism PCR (RFLP-PCR) for species identification.

RESULTS: Antibodies to Chlamydiaceae were detected in 30/39 (77%) and 50/63 (79%) buffalo without and with signs of reproductive disease, respectively, and 10/20 (50%) and 39/53 (74%) of cows with and without signs of reproductive disease, respectively. Positive samples from PCR analysis were identified in 31/39 (79%) and 37/63 (59%) buffalo without and with signs of reproductive disease, respectively, and 12/20 (60%) and 46/53 (89%) of cows without and with signs of reproductive disease, respectively. Using RFLP-PCR, 57/68 (84%) of samples from buffalo, and 47/58 (81%) from cows, were identified as Chlamydophila psittaci and the reminder as Cp. abortus. From the CFT and PCR results there was no significant difference in the prevalence of positive samples between species, or between animals without or with signs of reproductive disease.

CONCLUSION: The presence of anti-Chlamydiaceae antibodies in 77% of the animals with signs of reproductive disease and the detection of Chlamydiaceae in 72% of vaginal swabs of the animals suggest a pathogenic role by Chlamydiaceae in riverine buffalo and cows. The main Chlamydiaceae found in the genital tract of cattle in Egypt were Cp. psittaci and Cp. abortus.

CLINICAL RELEVANCE: Chlamydophila spp. should be included in diagnostic algorithms for reproductive disorders, in order to assess the real burden of Chlamydophila associated disease in buffalo and cattle and to evaluate the potential value of vaccines.  相似文献   

9.
Chlamydophila felis (C. felis), feline herpesvirus-1 (FHV-1) and feline calicivirus (FCV) were detected in 39 (59.1%), 11 (16.7%) and 14 (21.2%) cats respectively, from 66 domestic cats presented with conjunctivitis and upper respiratory tract disease (URTD) in 9 prefectures of Japan. Dual and multiple infections were found in 7 (10.6%) cats with both C. felis and FHV-1, 10 (15.2%) cats with both C. felis and FCV, and 1 (1.5%) cat with all three agents. C. felis was isolated from 11 (28.2%) of 39 PCR positive cats. Antigenic difference was found in a 96 kDa protein of our isolates and Fe/145 strain isolated in USA. In conclusion, C. felis is the most common agent of feline conjunctivitis and URTD, and the coinfection with C. felis, FHV-1 and FCV are also common in cats in Japan.  相似文献   

10.
Objective To investigate how different sampling techniques affect detection of DNA from feline herpes virus Type 1 (FHV-1), Chlamydophila felis and Mycoplasma felis and to study the correlation between positive test results and clinical signs in cats. Animals Fifty-one cats; 24 with ocular signs and 27 healthy control cats. Procedures Samples were collected from all cats using cotton swabs, conjunctival and corneal biopsies, and corneal scrapings. Samples were analyzed for presence of FHV-1, C. felis, M. felis, and feline DNA, defined by 28S rDNA, by using real-time PCR. Results In affected cats, FHV-1 was detected in only one cat; C. felis and M. felis were not detected in any affected cats. None of the three organisms was detected in any control cats. Feline DNA was demonstrated in all conjunctival samples, in 82% of corneal swabs, 92% of corneal scrapings, and 100% of keratectomy samples. Conclusions Because of the generally low detection rate for FHV-1, C. felis, and M. felis DNA in this study, differences regarding sampling technique could not be determined and correlation between positive test results and degree of clinical signs could not be made. Detection of feline DNA in most samples irrespective of sampling technique, suggests a low prevalence of FHV-1, C. felis and M. felis in this population of cats.  相似文献   

11.
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12.
The purpose of this study was to determine if Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' exist in Korea. Three hundreds and thirty one feral cats were evaluated by using PCR assay targeting 16S rRNA gene sequence. Fourteen cats (4.2%) were positive for M. haemofelis, 34 cats (10.3%) were positive for 'Candidatus M. haemominutum' and 18 cats (5.4%) were positive for both species. Partial 16S rRNA gene sequences were closely (>98%) related to those from other countries. This is the first molecular detection of feline hemoplasmas in Korea.  相似文献   

13.
The prevalence of Chlamydophila felis and feline herpesvirus 1 (FHV-1) infection in cats with conjunctivitis in northern Italy was investigated by conventional polymerase chain reaction (PCR) testing. In cats with conjunctivitis, C felis and FHV-1 were detected in 14 of 70 (20%) and in 23 of 70 (33%) animals, respectively. None of the 35 control cats were positive for C felis, whereas 7 (20%) of these cats were positive for FHV-1. Mixed infections were present in 5 of 70 cats (7%). Cats positive for C felis were significantly younger than control animals (P = .02), whereas no significant age differences were observed between FHV-1-positive cats and control cats (P = .41) or between FHV-1-positive animals and C felis-positive animals (P = .16). Cats sampled during acute-phase conjunctivitis were also investigated for the presence of C felis by conjunctival scrapings. In this acute phase, substantial agreement was found when comparing the results of the 2 methods (K = .80). The association between PCR results and conjunctivitis was evaluated for the 2 pathogens. The presence of C felis was significantly associated with conjunctivitis (P = .004), whereas the detection of FHV-1 did not significantly correlate with the clinical sign (P = .25), suggesting that, by itself. PCR is not suitable for the diagnosis of FHV-1-related conjunctivitis.  相似文献   

14.
Mycoplasma felis is associated with conjunctivitis and respiratory disease in domestic cats. Currently no rapid diagnostic test is available for the detection of M. felis in clinical samples that does not rely on prior cultivation of the organism. The objective of this study was to determine whether a polymerase chain reaction (PCR) based upon the 16S/23S rRNA intergenic spacer sequence is suitable for the identification of M. felis directly in feline clinical samples. The high conservation between the 16S/23S rRNA intergenic spacers (IGS) of differing isolates of M. felis was established by sequence analysis and a PCR was developed to this region by comparison to IGS of other mycoplasmas. The PCR was found to be highly specific for M. felis and further PCR analysis on clinical samples showed the PCR to be highly sensitive and more rapid than the other methods of identification currently available.  相似文献   

15.
Background: ‘Candidatus Mycoplasma turicensis’ (CMtc) is a hemotrophic bacterial species that can, alone or in combination, induce anemia in cats. The diagnostic test of choice for hemoplasma infections is PCR. Conventional PCR assays have been developed for the detection of Mycoplasma haemofelis (Mhf) and ‘Candidatus M. haemominutum’ (CMhm) but not for CMtc. Although real‐time PCR assays have been reported for all of the feline hemoplasmas, the expense of necessary instrumentation precludes its use in Brazil and many other countries. Objectives: The goals of this study were to develop and optimize a conventional PCR assay to diagnose CMtc using an internal control to detect false‐negative results, and to evaluate the occurrence of CMtc infection in domestic cats from Brazil. Methods: Species‐specific primers were designed and a PCR assay was developed for the detection of CMtc 16S rDNA in cat blood. Sensitivity was determined by serial 10‐fold dilutions of plasmid and DNA extracted from blood from an experimentally infected cat. EDTA blood samples from 373 cats were collected. DNA was extracted using a silica‐based protocol and tested using the PCR assay. Results: Primer concentration, annealing temperature, and MgCl2 concentration were optimized in the presence and absence of the internal control. Two samples negative for the internal control were excluded. Of the remaining 371 samples (117 healthy and 254 unhealthy cats), 17 (4.6%) were positive for CMtc. Conclusion: These results demonstrate the utility of an optimized PCR assay to detect CMtc in feline blood samples. We also report for the first time the prevalence of CMtc infection in domestic cats in Brazil.  相似文献   

16.
Chlamydiae are an important cause of acute and chronic conjunctivitis in cats. Until recently, only one organism was thought to infect cats, Chlamydophila felis (previously Chlamydia psittaci var. felis). Recently, other Chlamydia-like organisms belonging to the family Parachlamydiaceae, which comprises organisms that reside and proliferate within free-living amoeba, have been identified in cats with neutrophilic and eosinophilic conjunctivitis. The relative importance of these organisms and their amoebic hosts requires investigation. There is also weak evidence that chlamydiae may also be capable of causing reproductive tract disease and lameness in cats. Diagnosis of chlamydial conjunctivitis requires use of specialized culture techniques or the polymerase chain reaction. The antibiotic of choice to treat these infections is doxycycline; azithromycin is less effective. All cats in the household should be treated simultaneously. The zoonotic potential of these organisms appears low, but some precaution is warranted when handling affected cats.  相似文献   

17.
The aim of the present study was to analyse the occurrence of chlamydiae in several mammalian host species. Clinical samples that previously tested positive in a Chlamydiaceae-specific real-time PCR were retested using six species-specific real-time PCR assays to identify the chlamydial species involved. Chlamydophila (Cp.) abortus was the agent most frequently found in cattle, sheep, horses, goats, and pigs. Detection in cattle of Cp. psittaci (11% of samples) and Chlamydia (C.) suis (9%), as well as Cp. psittaci in a goat sample was somewhat unexpected. DNA of two different chlamydiae was identified in 56 (12.7%) of 440 samples tested. Cp. felis was the predominant species found in cats, while in guinea pigs and rabbits only Cp. caviae was detected. Interestingly, the latter two pathogens were also identified in samples from dogs. The data show that mixed chlamydial infections are not rare and suggest an extended host range of individual species.  相似文献   

18.
The prevalence of Chlamydia psittaci infection in household, feral and farm cats in Britain was investigated. Chlamydia were isolated from 30 per cent of conjunctival swabs collected from 753 household cats with conjunctivitis. The prevalence of active chlamydial infection was highest in cats in the age group five weeks to nine months. Males were more frequently infected than females. Cats with chlamydial conjunctivitis usually had antibody titres greater than 1024 as assessed by indirect immunofluorescence. Chlamydia appeared to be endemic in two out of three feral cat colonies on the basis of serological evidence and occasional isolations. Cats on 10 of 22 sheep farms (45 per cent) had serological evidence of chlamydial infection, and this was confirmed on two farms by isolation of the organism from conjunctival and, or, rectal swabs. This is the first survey of infection with Chlamydia psittaci in cat populations in Britain.  相似文献   

19.
Reports of canine chlamydiosis are infrequent, possibly because the pathogen is rarely considered to be a cause of disease in dogs. This report presents details of Chlamydophila psittaci infection in four bitches with recurrent keratoconjunctivitis, severe respiratory distress and reduced litter size (up to 50% stillborn or non-viable puppies) in a small dog-breeding facility in Germany. Cell culture and immunofluorescence examination of conjunctival, nasal and pharyngeal swabs revealed chlamydial inclusions. PCR and sequencing of ompA amplification products confirmed the presence of Cp. psittaci genotype C. The zoonotic potential of the pathogen was illustrated by evidence of disease in two children that lived on the premises with the infected dogs. There was circumstantial evidence to suggest infection of dogs and humans may have followed the introduction of two canaries and a parrot to the household. The persistent nature of the chlamydial infection suggests that dogs may be reservoirs of Cp. psittaci, but this putative role and whether or not dogs shed the pathogen require further investigation.  相似文献   

20.
The crossing of host species barriers, through the spreading populations of introduced pet animals that become established in the wild, sets the stage for zoonotic pathogen (re)emergence. A literature review on pathogens that are hosted by the ring-necked parakeet (Psittacula krameri), a worldwide introduced pet, highlighted local infections of captive birds by chlamydial agents with high sanitary risk for human health in its introduced range. We searched for these pathogens through cloacal swabs collected from 85 individuals in an invasive population established in the suburban areas of Paris (Ile-de-France) from 5 localities during the winter seasons between 2011 and 2014. Based on quantitative PCR analysis, Chlamydiaceae shedding was detected at too low levels for species identification in 5 birds, but 1 parakeet (found dead) was positive for Chlamydiaceae typed as Chlamydia avium. The only known hosts recorded for C. avium in Europe are feral pigeons (Columba livia) and captive psittacines. This result raises the question of the sanitary risks associated with new pathogen transmission from exotic pets released in the wild, which could locally affect birds and potentially people who feed birds.  相似文献   

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