Co-infection with Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in three cats from Brazil

The two most common haemotropic Mycoplasma of cats, Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' have been identified using molecular techniques in all continents, except Antarctica. We report the first molecular characterization in South America of a dual infection with M haemofelis and 'Candidatus Mycoplasma haemominutum' in three domestic cats. The 16S ribosomal RNA gene was amplified in three anaemic cats in which haemoplasma organisms were seen attached to the erythrocytes in the peripheral blood smear. Bands of the expected size for M haemofelis and 'Candidatus Mycoplasma haemominutum' were observed in all three cats. The 393 bp segment of one of the amplicons had a similarity value of 100% to M haemofelis, whereas the other amplicon, a 192 bp segment, was 100% similar to 'Candidatus Mycoplasma haemominutum'. After diagnosis, two cats received blood transfusion and they were all treated with doxycycline. All three cats recovered uneventfully.     

Occurrence of 'Candidatus Mycoplasma turicensis' infection in domestic cats in Japan

We have examined the presence of hemoplasmas, hemotropic mycoplasmas, among domestic cats (Felis catus) in Japan by using a species specific PCR, and found 'Candidatus Mycoplasma turicensis', a recently recognized hemoplasma species. A total of 60 feline blood samples collected in 2004 and 2005 were subjected to PCR amplification for the detection of Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis'. Six blood samples collected from domestic cats were found infected with the 'Candidatus M. turicensis'. All of them are also infected with other species of hemoplasmas, M. haemofelis and/or 'Candidatus M. haemominutum'. This is the first to demonstrate 'Candidatus M. turicensis' infections among cat population in Japan.     

Molecular detection of feline hemoplasmas in feral cats in Korea

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.     

Prevalence of selected infectious disease agents in cats from Arizona

The objective of this study was to use polymerase chain reaction (PCR) assays to determine the prevalence of Ehrlichia species, Anaplasma phagocytophilum, Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and Bartonella species from feral and relinquished cats in Phoenix and Nogales, Arizona. DNA from one or more of the organisms was amplified from 31 of 112 blood samples (27.7%). DNA consistent with Bartonella clarridgeiae 15 (13.4%), Bartonella henselae 14 (12.5%), 'Candidatus M haemominutum' 9 (8.0%), and M haemofelis 5 (4.5%) were detected. DNA of Ehrlichia species, Neorickettsia risticii, or A phagocytophilum was not amplified. Failure to amplify DNA of A phagocytophilum may relate to the absence of appropriate tick vectors. Failure to amplify Ehrlichia species DNA suggests that cats were not exposed, exposed but not infected, or infected but the DNA was not detected by the PCR assay used in this study. The Bartonella species and hemoplasma results suggest flea control should be maintained.     

Prevalence of DNA of Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum,' Anaplasma phagocytophilum, and species of Bartonella, Neorickettsia, and Ehrlichia in cats used as blood donors in the United States

OBJECTIVE: To identify the prevalence of DNA of Mycoplasma haemofelis; 'Candidatus Mycoplasma haemominutum'; Anaplasma phagocytophilum; and species of Bartonella, Neorickettsia, and Ehrlichia in blood of cats used as blood donors in the United States. DESIGN: Prospective study. ANIMALS: 146 cats that were active blood donors. PROCEDURES: Environmental history was requested for each blood-donor cat from which a blood sample (mixed with EDTA) was available. Polymerase chain reaction assays capable of amplifying the DNA of the microorganisms of interest following DNA extraction from blood were performed. RESULTS: Overall, DNA of one or more of the infectious agents was detected in blood samples from 16 of 146 (11%) feline blood donors. Twenty-eight laboratory-reared cats housed in a teaching hospital had negative results for DNA of all organisms investigated. The DNA of at least 1 infectious agent was amplified from blood samples collected from 16 of 118 (13.6%) community-source cats; assay results were positive for 'Candidatus M haemominutum,' M haemofelis, or Bartonella henselae alone or in various combinations. Of the community-source cats allowed outdoors (n = 61) or with known flea exposure (44), DNA for a hemoplasma or B henselae was detected in 21.3% and 22.7%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: When community-source cats, cats allowed outdoors, or cats exposed to fleas are to be used as blood donors, they should be regularly assessed for infection with M haemofelis, 'Candidatus M haemominutum,' and Bartonella spp, and flea-control treatment should be regularly provided.     

Use of a PCR assay to assess the prevalence and risk factors for Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in cats in the United Kingdom

Blood samples from 426 healthy and sick cats in the UK were tested in a PCR assay for 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemofelis (basonym Haemobartonella felis). Seventy-two of the cats (16.9 per cent) were positive for 'Candidatus M. haemominutum' alone, six (1.4 per cent) were positive for M. haemofelis alone and one (0.2 per cent) was positive for both. Logistic regression analysis indicated that older male cats were significantly more likely to be infected with 'Candidatus M. haemominutum', but there was no significant association between it and any of the haematological variables measured. M. haemofelis infection was uncommon in the anaemic cats sampled, and there were too few positive cases for multivariable analysis to be performed for M. haemofelis-positive status.     

Feline hemoplasmas in Switzerland: identification of a novel species, diagnosis, prevalence, and clinical importance

Two feline hemotropic mycoplasma spp. (aka hemoplasma) have previously been recognized. We recently discovered a third novel species in a cat with hemolytic anemia, designated 'Candidatus Mycoplasma turicensis', which is closely related to rodent haemoplasmas. This novel species induced anemia after experimental transmission to two SPF cats. Three quantitative real-time PCR assays were newly designed and applied to an epidemiological study surveying the Swiss pet cat population. Blood samples from 713 healthy and ill cats were analyzed. Up to 104 parameters per cat (detailed questionnaire, case history, laboratory parameters and retroviral infections) were evaluated. 'Candidatus Mycoplasma haemominutum' infection was more prevalent (8.5%) than Mycoplasma haemofelis (0.5%) and 'Candidatus Mycoplasma turicensis' (1%). Hemoplasma infections were associated with male gender, outdoor access, and old age, but not with disease or anemia. Infections were more frequently found in the South and West of Switzerland. Several hemoplasma infected cats, some acutely infected, others co-infected with FIV or FeLV, showed hemolytic anemia indicating that additional factors might play a role in the pathogenesis of the disease.     

Prevalence and risk factors for hemoplasmas in domestic cats naturally infected with feline immunodeficiency virus and/or feline leukemia virus in Rio de Janeiro--Brazil

The aim of this study was to determine the prevalence and risk factors for Mycoplasma haemofelis (Mhf) and 'Candidatus Mycoplasma haemominutum' (Mhm) infections in domestic cats tested for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Based on serological testing, cats were grouped as i) FIV-positive (n=25); ii) FeLV-positive (n=39); iii) FIV/FeLV-positive (n=8); and iv) FIV/FeLV-negative (n=77). Complete blood counts were followed by DNA extraction, species-specific polymerase chain reaction (16S rRNA gene) for Mhf and Mhm and Southern blotting for all animals. Mhf DNA was found in 4.0, 2.6, 12.5 and 7.8% of the cats from groups i, ii, iii and iv, respectively, while 32, 5.1, 50 and 5.2% of these animals had an Mhm infection. Cats with FIV (OR=4.25, P=0.009) and both FIV and FeLV (OR=7.56, P=0.014) were at greater risk of being hemoplasma infected than retroviral-negative cats, mainly due to Mhm infection (OR=8.59, P=0.001 and OR=18.25, P=0.001, respectively). Among pure-breed cats, FIV-positive status was associated with hemoplasma infection (OR 45.0, P=0.001).     

Diagnosis of feline haemoplasma infection using a real-time PCR assay

Haemobartonella felis has been reclassified within the genus Mycoplasma as Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum', collectively referred to as the feline haemoplasmas. A total of 78 cats from the Johannesburg area that had blood samples submitted to a private veterinary laboratory were tested using a real-time polymerase chain reaction (PCR) assay able to detect and distinguish the two feline haemoplasma (basonym Haemobartonella) species. All samples had been diagnosed with haemoplasma infection by cytological examination of blood smears. Statistical analysis was performed to evaluate associations between haemoplasma status, age, and haematological and biochemical parameters. On PCR assay 43 cats (55%) were haemoplasma negative, 25 (32.1%) positive for 'Candidatus Mycoplasma haemominutum', 5 (6.4%) positive for Mycoplasma haemofelis and 5 (6.4%) positive for both species. Significant inverse correlation was found between the amount of M. haemofelis DNA present in the blood and the haematocrit value. Cats that were positive for M. haemofelis showed macrocytic regenerative anaemia, monocytosis and thrombocytopaenia. This report documents the existence of both haemoplasma species in cats in South Africa.     

Prevalence of Bartonella species, haemoplasma species, Ehrlichia species, Anaplasma phagocytophilum, and Neorickettsia risticii DNA in the blood of cats and their fleas in the United States

Ctenocephalides felis were killed and collected from 92 cats in Alabama, Maryland, and Texas. The fleas and blood from the corresponding cat were digested and assessed in polymerase chain reaction assays that amplify DNA of Ehrlichia species, Anaplasma phagocytophilum, Neorickettsia risticii, Mycoplasma haemofelis, 'Candidatus M haemominutum' and Bartonella species. DNA consistent with B henselae, B clarridgeiae, M haemofelis, or 'Candidatus M haemominutum' was commonly amplified from cats (60.9%) and their fleas (65.2%). Results of this study support the recommendation to maintain flea control on cats in endemic areas.     

Prevalence of Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum', Bartonella species, Ehrlichia species, and Anaplasma phagocytophilum DNA in the blood of cats with anemia

Hemoplasmas are known causes of anemia in some cats and some Bartonella species have been associated with anemia in people and in dogs. In this retrospective study, we used polymerase chain reaction (PCR) assays to determine the prevalence rates of Mycoplasma haemofelis, 'Candidatus M haemominutum', A phagocytophilum, Ehrlichia species, and Bartonella species DNA in the blood of cats with anemia and a control group of healthy cats. DNA of the organisms was amplified from 22 of 89 cats with anemia (24.7%) and 20 of 87 healthy cats (23.0%). DNA of a hemoplasma was amplified from 18 of 89 cats with anemia (20.2%) and 13 of 87 healthy cats (14.9%); DNA of a Bartonella species was amplified from five of 89 cats with anemia (5.6%) and seven of 87 healthy cats (8.0%). There were no statistically significant differences detected between groups.     

Molecular detection of Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' Infection in cats by direct PCR using whole blood without DNA extraction

Detection of hemotropic Mycoplasma spp. infection was attempted in cats by PCR using whole blood without DNA extraction. A total 46 of 54 (85%) cats with suspected Mycoplasma spp. infection showed a positive reaction, corresponding completely with the results of standard PCR testing. The direct PCR assay was sensitive enough to detect more than 0.0061% parasitemia for ;C. M. haemominutum' and 0.0075% parasitemia for M. haemofelis. These data indicate that the direct PCR assay might be sufficient for use as a tool in clinical examinations.     

Use of conventional and real-time polymerase chain reaction to determine the epidemiology of hemoplasma infections in anemic and nonanemic cats

BACKGROUND: The goals of this study were to develop and apply conventional (c) and real-time TaqMan polymerase chain reaction (PCR) assays for Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma haematoparvum' (Mhp), and 'Candidatus Mycoplasma haemominutum' (Mhm) to blood samples of cats to determine the epidemiology of these infections in cats. HYPOTHESIS: Cats are infected with >2 hemoplasma species, and organism load correlates with disease induced by these organisms. ANIMALS: Blood samples from 263 anemic and nonanemic cats were used. METHODS: A retrospective study was conducted. RESULTS: Forty-seven (18%) samples were positive. Three samples (1%) yielded 170 base pair cPCR products, 1 of which was positive for Mhf using real-time PCR. Forty-four samples (17%) yielded 193 base pair cPCR products, 40 of which were positive for Mhm using real-time PCR. Organism loads ranged from 375 X 10(6)/mL to 6.9 x 10(6)/mL of blood. Sequencing of cPCR products from samples testing negative using real-time PCR identified 2 Mhp-like sequences, 1 Mhm-like sequence, and 1 sequence resembling 'Candidatus Mycoplasma turicensis'. Cats infected with Mhm were less likely to be anemic than uninfected cats. Older age, outdoor exposure, feline immunodeficiency virus (FIV) seropositivity, cutaneous squamous cell carcinoma (SCC), and stomatitis were associated with Mhm infection. Cats from the Sacramento Valley were more often infected with Mhm than cats from the San Francisco bay area. CONCLUSIONS AND CLINICAL IMPORTANCE: Cats may be infected with 4 hemoplasma species. The association between Mhm infection, FIV, and SCC may reflect outdoor roaming status of infected cats. The clustered distribution of infection suggests an arthropod vector in transmission.     

Diagnosis of feline haemoplasma infection in Australian cats using a real-time PCR assay

A total of 147 cats from the Sydney area of Australia that had blood samples submitted to veterinary laboratories were tested using a real-time polymerase chain reaction (PCR) assay able to detect and distinguish the two feline haemoplasma species. This sample number included two cats diagnosed with feline haemoplasma infection by routine blood smear examination. Statistical analysis was performed to evaluate associations between haemoplasma infection, age, sex, breed, haematocrit (HCT) values and anaemia status. One hundred and six cats (72.1%) were negative. Thirty-four cats (23.1%) were positive for 'Candidatus M. haemominutum', six cats (4.1%) were positive for M. haemofelis and one cat (0.7%) was positive for both species. Older, male, non-pedigree cats, with lower HCT values were more likely to be infected with 'Candidatus M. haemominutum'. Significant inverse correlation was found between the amount of M. haemofelis DNA present in the blood and the HCT value. This report documents the existence of, and prevalence of, both haemoplasma species in a sample of cats in Australia and is the first to use quantitative real-time PCR in a prevalence study for haemoplasma infection.     

Prevalences of various hemoplasma species among cats in the United States with possible hemoplasmosis

OBJECTIVE: To determine prevalences of various hemoplasma species among cats in the United States with possible hemoplasmosis and identify risk factors for and clinicopathologic abnormalities associated with infection with each species. DESIGN: Cross-sectional study. Animals-310 cats with cytologic evidence of hemoplasmosis (n = 9) or acute or regenerative anemia (309). PROCEDURES: Blood samples were tested by means of a broad-spectrum conventional PCR assay for hemoplasma DNA and by means of 3 separate species-specific real-time PCR assays for DNA from "Candidatus Mycoplasma haemominutum" (Mhm), Mycoplasma haemofelis (Mhf), and "Candidatus Mycoplasma turicensis" (Mtc). RESULTS: Overall prevalences of Mhm, Mhf, and Mtc infection were 23.2% (72/310), 4.8% (15/310), and 6.5% (20/310), respectively. Mixed infections were detected in 20 (6.5%) cats. Cats infected with hemoplasmas were more likely to be male than were uninfected cats. Infection with FeLV or FIV was significantly associated with infection with Mhf. Compared with uninfected cats, cats infected with Mhf had higher reticulocyte counts, nucleated RBC counts, and mean corpuscular volume; cats infected with Mhm had higher mean corpuscular volume; and cats infected with Mtc had higher monocyte counts. CONCLUSIONS AND CLINICAL RELEVANCE: Results supported the suggestion that these 3 hemoplasma species commonly occur among cats in the United States and that pathogenicity of the 3 species varies.     

Use of real-time PCR to detect Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in the saliva and salivary glands of haemoplasma-infected cats

Feline haemoplasma infection can cause haemolytic anaemia. The natural method of transmission of haemoplasmas between cats is currently unknown but the nature of some of the risk factors for infection suggests that saliva may act as a mode of transmission. The aim of this study was to determine if Mycoplasma haemofelis (Mhf) and 'Candidatus Mycoplasma haemominutum' (CMhm) DNAs could be amplified from saliva and salivary gland samples collected from haemoplasma-infected cats.     

Detection and molecular characterization of feline hemoplasmas in wild felid species in Iran in the Middle East

Three feline hemoplasma species exist in felids: Mycoplasma haemofelis, ‘Candidatus Mycoplasma haemominutum’, and ‘Candidatus Mycoplasma turicensis’.The aims of the study were to determine the presence of, and molecularly characterize, any hemoplasmas in wild felids, including the endangered Persian leopard in Iran, the Middle East.Blood samples were collected from 19 wild felids, including three Persian leopards. Using species-specific hemoplasma PCRs and ELISA serological testing for feline leukaemia virus and feline immunodeficiency virus (FIV), two Persian leopards were found to be infected with ‘Ca. M. haemominutum’ and were seropositive for FIV. Partial 16S rRNA gene sequences were generated for these ‘Ca. M. haemominutum’ species and subsequent phylogenetic analysis revealed 97.70% to 99.45% sequence identity with those found in domestic cats from Iran and other countries.This study confirms the presence of ‘Ca. M. haemominutum’ and concurrent FIV antibody in wild felids in Iran. This represents the first report of hemoplasma in wild felids in the Middle East as well as the first report of infection in Persian leopards.     

Marbofloxacin for the treatment of experimentally induced Mycoplasma haemofelis infection in cats

BACKGROUND: Administration of tetracyclines or fluoroquinolones is associated with improvement in clinical and laboratory abnormalities in cats infected with Mycoplasma haemofelis. No treatment protocol has consistently eliminated the organism, and antimicrobial susceptibility may vary among M. haemofelis isolates. Continued search for effective therapies is warranted. HYPOTHESIS: Marbofloxacin administered at the onset of clinical illness will be safe and effective for the treatment of M. haemofelis. ANIMALS: Fourteen young adult, laboratory-reared cats housed together in a specific pathogen-free facility. METHODS: Twelve cats were inoculated IV with 2.0 mL of blood from 2 M. haemofelis positive cats. Clinical parameters were assessed daily. CBC and hemoplasma polymerase chain reaction (PCR) assay were performed before inoculation, weekly for 1-3 weeks postinoculation (PI) and twice weekly 3-6 weeks PI. Treatment with marbofloxacin (2.75 mg/kg PO daily for 14 days) was initiated in 6 randomly selected cats when PCV was <30% or fever was >102.5 degrees F (39.2 degrees C). Cats that were PCR positive on day 7 of therapy were treated for 28 days. Cats that were PCR negative on day 42 PI were treated with 20 mg/kg methylprednisolone acetate IM on day 50 PI. RESULTS: Significant differences between groups on some days after inoculation included higher PCV and red blood cell counts, lower mean cell volume, and higher mean cell hemoglobin content in marbofloxacin-treated cats. No differences in PCR assay results were noted between groups. CONCLUSIONS AND CLINICAL IMPORTANCE: Marbofloxacin was safe and resulted in more rapid hematologic improvement in M. haemofelis-infected cats, but did not change clinical scores and did not consistently eliminate infection.     

The prevalence of Bartonella, hemoplasma, and Rickettsia felis infections in domestic cats and in cat fleas in Ontario

The prevalence of persistent bacteremic Bartonella spp. and hemoplasma infections was determined in healthy pet cats in Ontario. Blood samples from healthy cats sent to a diagnostic laboratory for routine health assessment over the course of 1 y were tested for Bartonella spp. using both polymerase chain reaction (PCR) and blood culture, and for the presence of hemoplasma by PCR. The overall prevalence of Bartonella spp. by PCR and by culture combined was 4.3% (28/646) [3.7% (24/646) Bartonella henselae, 0.6% (4/646) Bartonella clarridgeiae]. The novel B. henselae PCR developed for this study demonstrated nearly twice the sensitivity of bacterial isolation. The overall prevalence of hemoplasma was 4% (30/742) [3.3% (25/742) Candidatus Mycoplasma haemominutum, 0.7% (5/742) Mycoplasma haemofelis]. There was no significant difference between the prevalence of infection by season or by age (≤ 2 y, > 2 y). Candidatus Mycoplasma turicensis was identified, for the first time in Canada, in 1 cat. The prevalence of Bartonella (58%) and hemoplasma (47% M. haemofelis, 13% M. haemominutum) in blood from a small sampling (n = 45) of stray cats was considerably higher than that found in healthy pet cats.     

Acute phase response to Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' infection in FIV-infected and non-FIV-infected cats

The pathogenicity of Haemoplasma spp. in cats varies with 'Candidatus Mycoplasma haemominutum' (CMhm) causing subclinical infection while Mycoplasma haemofelis (Mhf) often induces haemolytic anaemia. The aims of this study were to characterise the acute phase response (APR) of the cat to experimental infection with Mhf or CMhm, and to determine whether chronic feline immunodeficiency virus (FIV) infection influences this response. The acute phase proteins serum amyloid A (SAA), haptoglobin (Hp) and α-1-acid glycoprotein (AGP) concentrations were measured pre-infection and every 7-14days up to day 100 post-infection (pi) in cats infected with either Mhf or CMhm. Half of each group of cats (6/12) were chronically and subclinically infected with FIV. Marbofloxacin treatment was given on days 16-44 pi to half of the Mhf-infected cats, and on days 49-77 pi to half of the CMhm-infected cats. FIV-infected animals had significantly lower AGP concentrations, and significantly greater Hp concentrations than non-FIV-infected cats when infected with CMhm and Mhf, respectively. Both CMhm and Mhf infection were associated with significant increases in SAA concentrations, while AGP concentrations were only significantly increased by Mhf infection. Mhf-infected cats had significantly greater SAA concentrations than CMhm-infected animals. Both Mhf and CMhm infections were associated with an APR, with Mhf infection inducing a greater response. Chronic FIV infection appeared to modify the APR, which varied with the infecting Haemoplasma species.