Cytauxzoon felis infections are present in bobcats (Lynx rufus) in a region where cytauxzoonosis is not recognized in domestic cats

This study was performed to determine the prevalence of Cytauxzoon felis (C. felis) infections in bobcats (Lynx rufus) from a region where C. felis is recognized in domestic cats, North Carolina (NC), and a region where C. felis is not recognized in domestic cats, Pennsylvania (PA). Samples from NC (n=32) were obtained post-mortem via cardiac puncture from legally trapped bobcats. Samples from PA (n=70) were collected post-mortem onto Nobuto blood collecting strips by the PA Game Commission. Each sample was tested using a C. felis specific PCR assay as well as a PCR assay targeting host DNA to rule out the presence of PCR inhibitors. Three samples were excluded due to the presence of PCR inhibitors. Thirty-three percent (10/30) of the samples from NC and 7% (5/69) of the samples from PA tested positive for the presence of C. felis. The proportion of C. felis positive bobcats from NC was significantly different than that from PA (P<0.005). Despite the lower prevalence of C. felis infections in bobcats from PA this finding is unique and indicates the potential for C. felis infections in domestic cats in the northeastern USA if the appropriate tick vectors are present. Veterinary practitioners in PA should be on alert for cytauxzoonosis in domestic cats. Further studies about the epidemiology and transmission of C. felis infections among both domestic cats and bobcats are needed.     

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.     

An etiological investigation of domestic cats with conjunctivitis and upper respiratory tract disease in Japan

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.     

A review of bacterial pathogens in Ctenocephalides felis in New Zealand

The cat flea, Ctenocephalides felis, is the recognised vector of Bartonella henselae, B. clarridgeiae and Rickettsia felis. Although these Gram-negative bacteria were only described in the last decade, they are already known to cause a variety of diseases in people, particularly children and the immunosuppressed. Such diseases include cat-scratch disease, bacillary angiomatosis, endocarditis, bacteraemia, encephalopathy, neuroretinitis, osteomyelitis and peliosis hepatis. Although most infections in cats and dogs appear to be subclinical, recent studies have provided growing evidence that the bartonellas can also cause serious problems in pets, including hepatitis, endocarditis, central nervous system (CNS) signs, lymphadenopathy, uveitis, cataracts and reproductive failure. In 2004, DNA of B. henselae, B. clarridgeiae and R. felis was demonstrated in cat fleas from New Zealand and pets and their owners in the country are thus at risk of infection. While flea control programmes have traditionally been advocated by veterinarians to prevent pruritis and tapeworms in pets, they should now also be recommended to prevent infections with the new flea-borne bacterial pathogens. To raise awareness of the organisms amongst veterinarians and animal health workers, this review describes: the biology of the organisms; clinical and laboratory features of infections in cats, dogs and people; diagnosis; and possible treatments and control of infections with these organisms.     

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.     

Efficacy of pradofloxacin in cats with feline upper respiratory tract disease due to Chlamydophila felis or Mycoplasma infections

BACKGROUND: Upper respiratory tract disease (URTD) of cats is caused by a number of pathogens, including Chlamydophila felis and Mycoplasma spp. For effective treatment of both infections, doxycycline and enrofloxacin are recommended, but adverse effects limit their use in cats. HYPOTHESIS: That the fluoroquinolone pradofloxacin is effective against C. felis and Mycoplasma infection in cats with URTD or conjunctivitis. ANIMALS: Thirty-nine cats with signs of URTD or conjunctivitis. METHODS: Placebo-controlled, double-blind clinical trial. Cats were randomly entered into 1 of 2 treatment groups: treated PO with either 5 mg/kg pradofloxacin q24h or 5 mg/kg doxycycline q12h for 42 consecutive days. Changes in health status and clinical scores were evaluated. The presence of C. felis and Mycoplasma spp. was determined by quantitative polymerase chain reaction (PCR) and nested PCR of conjunctival swabs, respectively. RESULTS: At the beginning of the study, C. felis and Mycoplasma spp. were detected in 23 and 20 cats, respectively. Cats of both groups responded rapidly with a marked improvement in clinical signs within the 1st week. During treatment with either drug, C. felis DNA copy number declined quickly. Complete elimination of Mycoplasma spp. was achieved in both groups; however, whereas all cats receiving doxycycline eliminated C. felis, 4 cats treated with pradofloxacin remained PCR-positive. CONCLUSION AND CLINICAL IMPORTANCE: This study demonstrates that both pradofloxacin and doxycycline have good efficacy against C. felis and Mycoplasma spp., resulting in a marked improvement of clinical signs. However, C. felis DNA remained in some cats after treatment with pradofloxacin, suggesting that infection might not have been eliminated.     

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.     

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.     

Use of a polymerase chain reaction assay to detect and differentiate two strains of Haemobartonella felis in naturally infected cats

OBJECTIVE: To develop a polymerase chain reaction (PCR) assay that detects and differentiates the Ohio strain of Haemobartonella felis (H. felis-OH) and the California strain of H. felis (H. felis-CA) and to apply the assay to blood samples from cats with and without suspected haemobartonellosis (suspect and control cats, respectively). SAMPLE POPULATION: 220 blood samples were examined; 82 were from suspect cats, and 138 were from control cats. PROCEDURE: A PCR assay was designed to detect and differentiate H. felis-OH and H. felis-CA. RESULTS: On the basis of PCR assay results, the overall prevalence of H. felis infection was 19.5% (43/220). Suspect cats (28.0%; 23/82) were significantly more likely than control cats (14.5%; 20/138) to be H. felis infected. Significantly greater numbers of suspect cats were H. felis-OH infected (12.2%, 9/82) or H. felis-OH and H. felis-CA infected (4.9%, 4/82) than control cats (0% [0/138] and 0.7% [1/138], respectively). Significantly more anemic cats were H. felis-OH infected (14.3%; 4/28) or H. felis-OH and H. felis-CA infected (7.1%; 2/28) than nonanemic cats (2.3% [3/128] and 0.8% [1/128], respectively). The PCR assay was more accurate than cytologic examination for detection of H. felis. CONCLUSIONS AND CLINICAL RELEVANCE: Haemobartonella felis infections are more common in cats than previously recognized. Haemobartonella felis-OH is apparently more pathogenic than H. felis-CA. The PCR assay is more accurate than cytologic examination for detection of H. felis infection and is an effective clinical tool for the detection and differentiation of both H. felis strains known to infect cats.     

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.     

Epidemiology of flea infestation of ruminants in Libya

The results of an epidemiological and clinical study of flea infestations of farm animals in northern Libya is reported. Of 12,130 sheep examined from 124 flocks, 150 sheep were found to be infested with fleas from 50 different flocks. Likewise 23 goats from 2981 examined, and 11 calves from 1124 cattle examined were infested No fleas were recovered from camels or horses. Of 1861 fleas recovered from farm livestock, 1857 were Ctenocephalides felis strongylus and 4 were Pulex irritans. Dogs from farms and local clinics were also examined. Eight farms dogs were found to be infested with P. irritans. Of 79 infested dogs examined in veterinary clinics, 53 were found infested with P. irritans, 11 with Ctenocephalides felis felis, 12 had a mixed infestation of P. irritans and C. felis felis. Single dogs had mixed infestation of P. irritans and C. canis; C. felis felis and C. canis; and P. irritans, C. felis felis and Echidnophaga gallinacia. C. felis felis was also found on 15 infested cats. C. felis felis was never found on large farm animals despite frequently sharing their environment with dogs or cats. Likewise C. felis stongylus was never isolated from dogs or cats. This is consistent with the hypothesis that C. felis strongylus has become adapted to large farm animals, whilst C. felis felis is better adapted to dogs and cats. However, four stockmen were found infested with a total of 176 C. felis strongylus, which suggests that this subspecies is also a potential zoonosis. A significantly higher proportion of intensive farms had animals with flea infestation compared to semi-intensive farms. Fleas were not found in nomadic herds. Infested farm animals often presented with excoriation, alopecia, pruritis and hyperkeratitis particularly on the lower limbs. These signs are consistent with the generation of flea-bite hypersensitivity.     

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.     

Pathogen carriage by the cat flea Ctenocephalides felis (Bouché) in the United Kingdom

The carriage of Bartonella, Rickettsia felis and haemoplasma species was investigated in cat fleas (Ctenocephalides felis) collected from 121 cats and dogs in the United Kingdom. DNA extracted from fleas was analysed using genus and species-specific PCR and amplicons were characterised using DNA sequencing. Fifty percent of flea samples were PCR positive for at least one pathogen. Twenty one percent were positive for R. felis, 17% for Bartonella henselae, 40% for haemoplasma species and 20% were infected with more than one of the pathogen species studied. It is clear from the results in this study that companion cats and dogs are commonly infested with Ct. felis carrying bacterial pathogens of significance to human and animal health. These findings raise the possibility that Ct. felis found on dogs and cats are a potential source of infection with such pathogens for humans.     

Efficacy of selamectin administered topically to pregnant and lactating female dogs in the treatment and prevention of adult roundworm (Toxocara canis) infections and flea (Ctenocephalides felis felis) infestations in the dams and their pups

The efficacy of selamectin in the treatment and prevention of naturally acquired Toxocara canis infections and experimentally induced flea (Ctenocephalides felis felis) infestations in dams and their suckling pups was evaluated by administering selamectin to the adult females only, approximately 40 and 10 days before parturition and 10 and 40 days after parturition. Unit doses of the commercial formulation of selamectin were administered to the dams to provide at least the minimum recommended dosage of 6mgkg(-1) (range, 6-12mgkg(-1)). Dams and their pups were housed in carpeted environments able to support the flea life cycle. Flea infestations were established initially by experimental infestation before treatment administration and by repeated re-infestation of dams at approximately weekly intervals throughout the study, which was completed 45 days after parturition. There were no adverse drug experiences related to treatment with selamectin and no treatment-related mortalities. Percentage reductions in geometric mean T. canis faecal egg counts for the selamectin-treated dams, compared with those receiving the negative-control treatment (vehicle only) were 99.7% at the end of the study (P=0.0001). Geometric mean faecal egg counts in pups from selamectin-treated females were reduced by > or =96% on the 24th and 34th days after birth (P=0.0001), and the number of adult worms recovered from the gastrointestinal tract of pups from selamectin-treated dams was reduced by 98.2% (P=0.0001), compared with that for pups from dams treated with the vehicle only. Percentage reductions in geometric mean flea counts for selamectin-treated dams and their pups, compared with vehicle-treated dams and their pups, were > or =99.8% (P=0.0001) and 100% (P=0.0001), respectively, throughout the study. Thus, selamectin administered topically at a minimum unit dosage of 6mgkg(-1) to dams with naturally acquired T. canis infections and experimentally induced C. felis infestations was safe and highly effective in the treatment, control, and prevention of adult T. canis infection and C. felis infestation affecting both the dams and their pups.     

Serological response to pgp3 protein in animal and human chlamydial infections

Specific antibodies to plasmid-encoded protein pgp3 are known to be encountered in human Chlamydia (C.) trachomatis infections. In order to verify whether antibodies to this protein could be developed in animals infected with plasmid-carrying chlamydial strains, 454 animal sera were examined using a home-made pgp3 protein ELISA and Western blots (WB) of recombinant pgp3 protein from Chlamydophila (Cp.) psittaci. Likewise, 50 human sera were tested by ELISA and WB of recombinant pgp3 from C. trachomatis. The reactivity against pgp3 protein was compared to the reactivity against chlamydial elementary bodies (EBs) detected by microimmunofluorescence (MIF) test. The presence of pgp3-specific antibodies was demonstrated in most ducks and pigeons with Cp. psittaci infection detected by MIF, as well as in the majority of symptomatic cats and pigs infected with Cp. felis and C. suis, respectively, which reacted at high titres to Cp. felis and C. suis EBs by MIF. Moreover, most of the sera collected from patients with C. trachomatis culture-confirmed infection and seropositive to C. trachomatis by MIF, presented antibodies specific to C. trachomatis pgp3 recombinant protein. Therefore, pgp3 protein could be a useful marker of chlamydial infections in animals, as well as in humans.     

Sequestration and phagocytosis of Haemobartonella felis in the spleen.

Spleens of two cats infected with Haemobartonella felis were examined by electron microscopy to determine the means by which the organism was sequestered in this organ. The principal means of sequestration occurred when H felis, located on the erythrocytes was removed by phagocytosis by a cordal macrophage, apparently preceded by the adhesion of extended processes of the macrophage to H felis. The second and least frequent means of removal of H felis was by pitting, a process that did not cause destruction of the host erythrocyte. The H felis was pitted from the parasitized erythrocyte when H felis passed through gaps between reticular cells or when the parasitized erythrocyte passed among the cytoplasmic processes of the reticular cells in the splenic cords. Some H felis were closely associated with the plasmalemma of cordal reticular cells and also were located in intracytoplasmic vacuoles of the cells without being influenced by the phagocytic process.     

Detection of severe fever with thrombocytopenia syndrome virus and other viruses in cats via unbiased next-generation sequencing

We used unbiased next-generation sequencing (NGS) to detect unknown viruses in cats. Serum or plasma samples were obtained from clinically ill cats with suspected acute viral infections. Nucleic acid was extracted from serum or plasma samples to construct a complementary DNA library for NGS. Comprehensive nucleotide sequencing analyses enabled detection of the genomes of various viruses, including the severe fever with thrombocytopenia syndrome virus, feline immunodeficiency virus, feline morbillivirus, parvovirus, and Torque teno felis virus. Our findings indicate that comprehensive nucleotide analyses of serum or plasma samples can be used to detect infections with unknown viruses in cats.     

Occurrence of Babesia felis and Babesia leo in various wild felid species and domestic cats in Southern Africa, based on reverse line blot analysis

Reverse line blot (RLB) is a hybridization assay that can be used to detect various blood parasites and differentiate between them. Results, using the RLB, showed that Babesia felis and Babesia leo occurred as single or mixed infections in various felid species, but most frequently in domestic cats and lions, respectively. Prevalence of infection in free-ranging cheetahs in Namibia was low (7, 5%), whereas 50% of free-ranging lions in South Africa and Swaziland were infected. A large number (52, 9%) of samples tested positive only for Babesia, neither B. felis nor B. leo. This could be an indication of at least one further, as yet undescribed, Babesia species in felids.     

Haemobartonella felis: recent developments in diagnosis and treatment

Haemobartonella felis is a pleomorphic uncultivated wall-less haemotrophic bacterial parasite. Phylogenetic analysis of 16S rRNA gene sequences from a number of isolates of H felis has demonstrated that these bacteria are most closely related to species in the genus Mycoplasma, and Haemobartonella and related organisms are currently being reclassified as Mollicutes. Diagnosis by cytological examination of blood smears has been problematic, but recent molecular studies have led to the development of sensitive polymerase chain reaction (PCR) assays for diagnosis. Such studies have also resulted in the recognition of two distinct strains of H felis, which are divided into different groups based on phylogenetic analysis. This evolutionary divergence between strains is accompanied by differences in pathogenecity. This review discusses new developments in the diagnosis and treatment of H felis, focusing on the use of, and interpretation of, PCR assays.