Detection of persistent Cytauxzoon felis infection by polymerase chain reaction in three asymptomatic domestic cats

Repeated polymerase chain reaction (PCR) testing of 3 asymptomatic domestic cats were positive for Cytauxzoon felis DNA, suggesting persistent infection. Two cats initially presented with clinical signs consistent with acute cytauxzoonosis and, in both cases, signs of illness resolved after treatment. Parasitemia was detected in peripheral blood smears from these cats upon presentation with illness and, at subsequent follow-up appointments, in the absence of clinical illness. Polymerase chain reaction analysis was positive for C. felis from blood sampled at each time point. A third cat, a housemate of a cat fatally infected with C. felis, was preventatively treated for infection at the time of the housemate cat's death. This contact cat, having never shown signs of clinical illness consistent with cytauxzoonosis infection, had no detectable parasitemia but was positive for C. felis on repeated PCR testing. Detection of asymptomatically infected cats allows for the possibility of a yet unrecognized population of infected domestic cats that may have the capacity to serve as an additional reservoir host for C. felis, altering the currently accepted paradigm of C. felis transmission to domestic cats through bobcats as the reservoir host. In cases of very low parasitemia, more sensitive means of parasite detection, such as PCR testing, may be necessary to detect infected cats. Increased detection of asymptomatically infected cats will aid in understanding the epidemiology of C. felis infection and enhance the ability to prevent this highly fatal infectious disease of domestic cats.     

Feline haemobartonellosis: clinical, haematological and pathological studies in natural infections and the relationship to infection with feline leukaemia virus

Haemobartonella felis infection was demonstrated in 38 cats which could be divided into four groups as follows: group A, feline leukaemia virus (FeLV) free cats with H felis infection alone; group B, FeLV free cats with H felis infection and other clinical conditions; group C, FeLV positive cats with H felis infection but no clinical manifestation of FeLV related or any other intercurrent disease; and group D, FeLV positive cats with H felis infection and clinical manifestations of FeLV related or other diseases. Cats in group A were healthy carriers of the infection and none was anaemic, whereas some in group B had clinical haemobartonellosis and anaemia. This anaemia was mainly mild, normocytic and normochromic. Most of the cats in group C and all in group D were more severely ill and anaemic, the anaemia usually being macrocytic and hypochromic. Splenomegaly occurred only in groups C and D. Treatment with tetracyclines did not eliminate H felis from any of the cats and blood transfusions were ineffective in promoting long term recovery from anaemia in cats with intercurrent H felis and FeLV infections. The findings in the cats in groups C and D were further compared with those in a fifth group of cats which were infected with FeLV but free of H felis.     

The detection of Cytauxzoon felis in apparently healthy free-roaming cats in the USA

Cytauxzoon felis typically causes fatal disease in domestic cats. Survival after infection and persistent parasitemia without clinical illness has been documented in a few cases. To our knowledge there are no prevalence studies of C. felis in domestic cats. The purpose of this study was to estimate the prevalence of C. felis infected cats that were presented to trap-neuter-return programs in Florida, North Carolina and Tennessee. Cats that were presented to trap-neuter-return programs were tested using a C. felis-specific PCR assay. A total of 961 domestic cats were tested (494 from Florida; 392 from North Carolina; 75 from Tennessee). Prevalence of C. felis infection in this population was 0.3%. Two cats from Florida and one cat from Tennessee tested positive for the presence of C. felis DNA. These amplicons were sequenced and confirmed to be C. felis. The cat from Tennessee was alive without evidence of illness 2 months post-surgery. The other two cats were alive 24 h post-surgery, but were then lost to follow-up. This is the first report documenting C. felis infections in free-roaming cats. Despite the low prevalence rate, the presence of apparently healthy infected free-roaming cats suggests that they may have the capacity to serve as an additional reservoir host for C. felis. Further investigations should evaluate the potential vector competence of domestic cats as well as the role of chronically infected cats in areas in which cytauxzoonosis appears hyperendemic.     

Is Chlamydophila felis a significant zoonotic pathogen?

PROBLEM: Chlamydophila felis is a common cause of conjunctivitis in cats and in some textbooks is listed as an important zoonotic pathogen. However, there are no published comprehensive reviews assessing the evidence supporting this. SEARCH AND ANALYSIS: Bibliographic databases and bibliographies of papers and textbooks were searched for all published cases of zoonotic disease associated with chlamydiosis in cats. All published case reports were reviewed to establish the quality of the evidence supporting the association between C. felis and zoonotic disease. RESULTS: There are only seven case reports, most of which were published before the development of assays capable of distinguishing C. felis from other chlamydial species, implicating this organism in zoonotic disease. None of the three cases of pneumonia and systemic disease described, two of which occurred in immunocompromised patients, can be unambiguously attributed to C. felis. Of the four cases of follicular keratoconjunctivitis described, only one, in an immunocompromised patient, could be unambiguously attributed to C. felis. CONCLUSION: While there is evidence that C. felis may occasionally cause keratoconjunctivitis in humans, there is little evidence to suggest it can cause serious systemic disease or pneumonia.     

Attempted transmission of Candidatus Mycoplasma haemominutum and Mycoplasma haemofelis by feeding cats infected Ctenocephalides felis

OBJECTIVE: To determine whether Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma haemominutum (Mhm) can be transmitted by ingestion of Mycoplasma-infected Ctenocephalides felis and by-products (feces, larvae, and eggs). ANIMALS: 10 cats. PROCEDURE: 3 cats were carriers of Mhf, and 1 was a carrier of Mhm. Six cats had negative results of PCR assay for Mhf and Mhm DNA. A chamber containing 100 C felis was bandaged to 2 Mhf carrier cats. Five days later, fleas and by-products were analyzed for Mycoplasma spp DNA. The remaining fleas and a sample of by-products were fed to 2 Mycoplasma-na?ve cats. A chamber containing 200 C felis was bandaged to the Mhm carrier cat. Five days later, fleas and by-products were analyzed for Mycoplasma spp DNA. The remaining fleas and a sample of by-products were fed to 2 Mycoplasma-na?ve cats. A chamber containing 200 C felis was bandaged to an Mhf carrier cat and Mhm-carrier cat. Three days later, fleas and by-products were analyzed for Mycoplasma spp DNA. The remaining fleas and a random sample of by products were fed to 4 Mycoplasma-na?ve cats. All cats were monitored for infection for >or=7 weeks. RESULTS: Uptake of Mhf and Mhm DNA into fleas and by-products was detected. None of the na?ve cats became infected. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that ingestion of Mycoplasma-infected C felis or by-products is not an important means of transmission for Mhf or Mhm.     

Transmission of feline leukaemia virus in the milk of a non-viraemic cat

The possibility of the transmission of feline leukaemia virus (FeLV) from latently infected cats was studied. Five female cats with latent infections were examined for evidence of transmission of the virus to their kittens. One of the cats infected members of four consecutive litters of kittens which subsequently became persistently viraemic and transmitted the virus to other susceptible kittens by contact. Shortly after birth its kittens were apparently FeLV-free since neither viral antigen nor infectious virus was detected in their blood and no virus was found in cell cultures made from aspirates of bone marrow. The kittens became viraemic from 45 days of age onwards at a time when their passively acquired colostral FeLV neutralising antibodies were no longer detectable. Transmission of the virus occurred via the milk since both FeLV antigen and infectious virus were found in milk samples taken six weeks after kittening and the virus was transmitted to a fostered kitten. Eleven weeks after the birth of the fourth litter the cat became viraemic. The intermittent presence of FeLV antigens detected by the Leukassay F test, but not infectious virus, in the plasma of this cat over the previous months and a low level of serum neutralising antibodies distinguished it from four other latently infected queens which did not transmit infection to their kittens. These factors may indicate a risk of milk transmission and reactivation of latent virus.     

Development and evaluation of a PCR assay for the detection of Cytauxzoon felis DNA in feline blood samples

Cytauxzoonosis is an emerging tick borne infectious disease of domestic cats in the United States, caused by the organism Cytauxzoon felis (C. felis). In naturally infected domestic cats the disease is almost always fatal. Currently there are no commercially available molecular or serologic tests to facilitate the antemortem diagnosis of C. felis infection. Clinical and pathological diagnosis of cytauxzoonosis is based on microscopic identification of parasites in tissues or on blood smears. We have developed and evaluated the sensitivity and specificity of a polymerase chain reaction (PCR) based assay for the diagnosis of C. felis infections in feline blood samples. The assay is sensitive enough to detect one copy of a cloned fragment of the C. felis 18S rRNA gene. This PCR assay can be used for the rapid clinical diagnosis of cytauxzoonosis and for epidemiological studies that will better define the geographic distribution of C. felis infection in cats.     

Cytauxzoon felis infection in cats in the mid-Atlantic states: 34 cases (1998-2004)

OBJECTIVE: To describe the demographic and clinical characteristics of feline cytauxzoonosis in the mid-Atlantic states and compare the Cytauxzoon felis 18S rRNA gene sequences from affected cats with sequences reported from affected cats in other regions. DESIGN: Retrospective case series. ANIMALS: 34 cats with C. felis infection. PROCEDURE: Medical records of cats in which C. felis infection was diagnosed from May 1998 through June 2004 were reviewed; data collected included signalment, month of diagnosis, geographic location, clinicopathologic abnormalities, medical treatments, outcome, and necropsy findings when applicable. Cytauxzoon felis DNA was amplified, cloned, and sequenced from 4 of these cats and compared with previously reported C. felis DNA sequences. RESULTS: Of 34 C. felis-infected cats, 28 resided in North Carolina, 3 resided in South Carolina, and 3 resided in Virginia; in 32 cats, a diagnosis of C. felis infection was made in April through September. Pancytopenia and icterus were the most common clinicopathologic abnormalities. Thirty-two cats either died or were euthanatized, and 2 cats survived. At 5 veterinary hospitals, multiple cases were identified, and 4 multicat households had > 1 cat infected with C. felis. The 18S rRNA gene sequences characterized in organisms obtained from 4 cats were nearly identical to C. felis DNA sequences reported from other US regions. CONCLUSIONS AND CLINICAL RELEVANCE: Data indicate that veterinarians in the mid-Atlantic region of the United States should consider C. felis infection in cats that become ill with fever, icterus, and pancytopenia or bicytopenia, especially in the spring and summer months.     

Feline chlamydiosis

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.     

A study of naturally occurring feline coronavirus infections in kittens.

Feline coronavirus is a common infection in cats, as indicated by the high prevalence of antibodies against the virus, especially in multicat households. Approximately 5 to 12 per cent of seropositive cats develop classical feline infectious peritonitis. A survey of kittens born into households of seropositive cats demonstrated the existence of healthy coronavirus carriers. Seronegative animals did not appear to excrete virus. No specific antibody titre could be linked to carrier status and some carrier cats subsequently became seronegative. The management of the kittens strongly influenced whether they became infected, and some degree of protection appeared to be conferred by maternally derived antibody. At present, feline infectious peritonitis virus and feline enteric coronavirus can only be differentiated by their different clinical histories in infected catteries. In this survey, cases of feline infectious peritonitis occurred in kittens from households where the initial presentation had been enteritis and vice versa. Therefore no difference in epidemiology could be found.     

Treatment of experimentally induced cytauxzoonosis in cats with parvaquone and buparvaquone

The efficacy of parvaquone (Clexon) and buparvaquone (Butalex) in treating experimentally induced feline cytauxzoonosis was explored. Domestic cats were inoculated subcutaneously with blood from a cat infected with Cytauxzoon felis and treated daily with either 20 or 30 mg kg-1 parvaquone, or 5 or 10 mg kg-1 buparvaquone, beginning on either the first day parasites were detected in peripheral blood, or 2 days after the onset of parasitemia. Fifteen cats were treated and all but one died due to the infection. Unexpectedly, one of two non-treated, infected control cats survived. Although parvaquone and buparvaquone are the treatments of choice for a related hemoprotozoan parasite causing theileriosis in African cattle, wer concluded that at the dosages and regimes tested, these drugs are not effective treatments for feline cytauxzoonosis. Blood from the two surviving cats was inoculated into naive cats and in these animals clinical disease or death were not observed. The latter two naive recipient cats were then inoculated with a lethal dose of viable, frozen C. felis and both died, thereby indicating that blood from surviving cats did not induce an infectious state that resulted in immunity. The two cats that survived the acute infection were subsequently challenged with a lethal inoculum of C. felis; they showed no clinical signs of cytauxzoonosis and were obviously immune to reinfection.     

New perspectives about Hemotrophic mycoplasma (formerly, Haemobartonella and Eperythrozoon species) infections in dogs and cats.

The new perspectives about hemotrophic mycoplasma infections in cats and dogs can be summarized as follows: Haemobartonella and Eperythrozoon species infecting the dog and cat have been reclassified as mycoplasmal parasites and given the names M haemofelis (Ohio or large form of H felis), M haemominutum (California or small form of H felis), and M haemocanis (H canis). The prevalence of hemotrophic mycoplasma infections in anemic cats in the United States is about 25% and usually involves M haemofelis. However, nonanemic cats may also be infected most commonly with M haemominutum. Chronic infections with hemotrophic mycoplasmas may promote myeloproliferative disorders in FeLV-infected cats. M haemocanis infection in dogs may be a widespread latent disease in kennel-raised dogs and is being investigated. The PCR assay is exquisitely sensitive for detection of M haemofelis and M haemominutum, and testing of blood donor cats and perhaps dogs should be done regularly. Fleas are involved in the transmission of M haemofelis to the cat, whereas R sanguines may be involved with transmission of M haemocanis to the dog. Treatment with doxycycline effectively controls acute infection in the cat and dog, and enrofloxacin may also be effective in the cat, but none of the antibiotics tested to date consistently clears the parasites.     

Experimental infection of domestic cats (Felis domesticus) with Cytauxzoon manul from Pallas' cats (Otocolobus manul)

In a random, blind study, six domestic cats were assigned to two treatment groups that received either sterile water or dexamethasone by subcutaneous injection prior to intravenous inoculation with Pallas' cat (Otocolobus manul) blood infected with Cytauxzoon manul. A seventh domestic cat served as a control and was inoculated only with sterile water. Cats were monitored for clinical signs consistent with cytauxzoonosis, and periodically screened for hemoparasitemia. All domestic cats (6/6) that received Pallas' cat blood infected with C. manul developed a low but detectible parasitemia by 9 days post-inoculation, yet remained clinically healthy. All domestic cats (7/7) were subsequently challenged with Cytauxzoon felis and developed clinical signs typical of cytauxzoonosis within 5 days post-challenge. Affected animals were euthanized and cytauxzoonosis was confirmed by histopathology. While inoculation of domestic cats with Pallas' cat blood infected with C. manul induced a parasitemia, it did not cause disease or provide protection against challenge with C. felis. Further studies are warranted to determine the potential for interspecies transmission and disease with C. manul.     

Evaluation of experimental transmission of Candidatus Mycoplasma haemominutum and Mycoplasma haemofelis by Ctenocephalides felis to cats

OBJECTIVE: To determine whether Ctenocephalides felis can transmit Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma haemominutum (Mhm) through hematophagous activity between cats. ANIMALS: 11 cats. PROCEDURE: 2 cats were carriers of either Mhf or Mhm. Nine cats had negative results via polymerase chain reaction (PCR) assay for Mhf and Mhm DNA; 3 of those cats were infected from the chronic carriers via i.v. inoculation of blood. At the time of maximum organism count for each of the Mycoplasma spp, 1 chamber containing 100 C felis was bandaged to the amplifier cats. Five days later, fleas, feces, larvae, or eggs from each chamber were analyzed for Mycoplasma spp DNA. Viable fleas from the chambers were allocated into new chambers (3 Mhm and 6 Mhf) and attached to na?ve cats for 5 days. Cats were monitored daily for clinical signs and weekly via CBC and PCR assay for infection with Mhf or Mhm for a minimum of 8 weeks. RESULTS: Uptake of Mhf and Mhm DNA into fleas, feces, and, potentially, eggs and larvae was detected. Of the na?ve cats fed on by Mhf-infected fleas, 1 cat transiently yielded positive PCR assay results for Mhf on 1 sampling date without clinical or hematologic changes consistent with Mhf infection. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that hematophagous transfer of Mhm and Mhf into fleas occurred and that C felis is a possible vector for Mhf via hematophagous activity.     

Using CF0218-ELISA to distinguish Chlamydophila felis-infected cats from vaccinated and uninfected domestic cats

Chlamydophila felis is a causative agent of acute and chronic conjunctivitis and pneumonia in cats. Cats can be vaccinated with killed or attenuated C. felis. However, current serodiagnostics cannot distinguish these cats from naturally infected cats. This causes difficulty of early diagnosis and seroepidemiological survey for C. felis. We previously reported that C. felis CF0218 can be used as a C. felis-infection-specific diagnostic antigen in experimentally infected and/or vaccinated cats. In this study, we evaluated an enzyme-linked immunosorbent assay using recombinant CF0218 as antigen (CF0218-ELISA) to detect anti-C. felis antibody in 714 sera of domestic cats whose histories of vaccination against C. felis are known. The 44 vaccinated cats were 93% negative using CF0218-ELISA; half of these scored positive by immunofluorescence assay (IFA) using C. felis-infected cells as antigen. The 670 non-vaccinated cats had CF0218-ELISA positivity rates that were statistically in agreement with IFA (18% vs. 21%). These results show that CF0218, which was identified as a C. felis-infection-specific antigen, is a useful serodiagnostic antigen to distinguish naturally C. felis-infected cats from vaccinated and non-infected cats.     

Pharmacokinetics of enrofloxacin and its efficacy in comparison with doxycycline in the treatment of Chlamydophila felis infection in cats with conjunctivitis

The concentrations of enrofloxacin were measured in the tears, saliva and serum of 14 cats with signs of upper respiratory tract infection and eight with no signs, after daily doses of 5 mg/kg. Enrofloxacin concentrations above the minimum inhibitory concentration of Chlamydophila felis were found in the saliva and tears of the cats with and without signs of upper respiratory tract infection. In a prospective randomised clinical trial, the efficacy of enrofloxacin against C. felis infection in cats with conjunctivitis was compared with the efficacy of doxycycline. Twenty-five cats were randomly assigned to treatment with either enrofloxacin or doxycycline for 14 days; 15 of the cats tested positive for C. felis by an immunofluorescent antibody test on conjunctival swabs. The two treatment groups showed equal improvements in the clinical signs of conjunctivitis and C. felis infection status; in each group three cats were still C. felis antigen-positive after the 14-day course of treatment, indicating a persistent infection. No side effects were observed in the cats treated with enrofloxacin.     

An enteric coronavirus infection of cats and its relationship to feline infectious peritonitis

An enteric coronavirus that is antigenically closely related to feline infectious peritonitis virus (FIPV) is ubiquitous in the cat population. This virus has been designated feline enteric coronavirus to differentiate it from FIPV. The virus is shed in the feces by many seropositive cats; in catteries it is a cause of inapparent to mildly severe enteritis in kittens 6 to 12 weeks of age. The virus may produce a more severe enteritis in young specific-pathogen-free kittens. Feline enteric coronavirus selectively infects the apical columnar epithelium of the intestinal villi, from the caudal part of the duodenum to the cecum. In severe infections, there are sloughing of the tips of the villi and villous atrophy. Many cats recovering from the disease remain carriers of the virus. Recovered cats, observed for 3 to 24 months, remained healthy and did not develop peritonitis, pleuritis, or granulomatous disease. The relationship of feline enteric coronavirus and FIPV was studied. Although the viruses were antigenically similar, they were distinctly different in their pathogenicities. The enteric coronavirus did not cause feline infectious peritonitis in coronavirus antibody-negative cats inoculated orally or intraperitoneally nor in coronavirus antibody-positive cats inoculated intraperitoneally or intratracheally. Serologic tests, using FIPV, canine coronavirus, and transmissible gastroenteritis virus of swine as substrate antigens in fluorescent antibody procedures may not accurately identify FIPV infection. These tests do not appear to distinguish between FIPV and this feline enteric coronavirus.     

Efficacy of azithromycin for the treatment of feline chlamydophilosis

The current recommended treatment for feline chlamydophilosis involves daily oral administration of antimicrobials to all cats within an affected group for a prolonged period of time (4-6 weeks). Not surprisingly, owner compliance can be poor resulting in apparent treatment failure. Recent anecdotal evidence, supported by its efficacy in the treatment of Chlamydia trachomatis infection in humans, has suggested that azithromycin may offer an alternative by allowing less frequent dosing for a shorter duration. A clinical trial was designed to evaluate the efficacy of azithromycin for the treatment of chlamydia (Chlamydophila felis) infection in cats. Whilst azithromycin, given at 10-15 mg/kg daily for 3 days and then twice weekly, provided a similar, rapid resolution of clinical signs and negative isolation scores as doxycycline, C felis was re-isolated in four out of the five cats treated. Furthermore, even daily administration of azithromycin to chronically infected cats was ineffective in clearing infection. The azithromycin protocols used here were therefore found to be unsuccessful in eliminating the carriage of this strain of C felis.     

Prevalence of Chlamydophila felis and feline herpesvirus 1 in cats with conjunctivitis in northern Italy

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.