Survival of Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in blood of cats used for transfusions

Blood transfusions are commonly administered to cats; associated risks include the transmission of various infectious diseases including Mycoplasma haemofelis (Mhf) and 'Candidatus Mycoplasma haemominutum' (Mhm). Blood transfusions in citrate-phosphate-dextrose-adenine (CPDA-1) solution are commonly administered immediately or stored for up to 1 month prior to administration. It is unknown whether Mhf or Mhm survive in this solution or temperature. The purpose of this study was to determine if Mhf or Mhm remain viable after storage in CPDA-1 for varying periods of time. The results provide evidence that transmission of hemoplasmas to na?ve cats occurs after administration of infected feline blood that has been stored in CPDA-1 solution for 1h (Mhf) and 1 week (Mhm). These findings support the recommendation that cats used as blood donors be screened for Mhf and Mhm infections by polymerase chain reaction (PCR) assay prior to use.     

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.     

Molecular survey of Mycoplasma haemofelis and 'Candidatus mycoplasma haemominutum' infection in cats in Yamaguchi and surrounding areas

A molecular survey of hemoplasma (Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum') in Yamaguchi Prefecture and surrounding areas was performed by using molecular methods. PCR-RFLP with HindIII revealed that 2 cats were infected with M. haemofelis, and 16 with 'C. Mycoplasma haemominutum' among 102 randomly selected cats. Partial 16S rRNA gene sequences of M. haemofelis and 'C. Mycoplasma haemominutum' determined in this study showed percent similarities of 98.3-99.8% and 96.4-100%, respectively, with those from other countries. Hemoplasma infections were more frequently detected in free-roaming cats than inside cats. Also, the status of FeLV infection was another significant risk factor for hemoplasma infection.     

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.     

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.     

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.     

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.     

Mycoplasma haemofelis and Mycoplasma haemominutum detection by polymerase chain reaction in cats from Saskatchewan and Alberta

Hemobartonellosis is caused by Mycoplasma haemofelis, previously known as Haemobartonella felis. Cats infected with this organism typically develop regenerative anemia. The related species Mycoplasma haemominutum may also cause anemia. The purposes of this study were to use polymerase chain reaction technology to determine if both organisms exist in naturally infected cats from Saskatchewan and Alberta, and to determine if disease manifestation corresponds to mycoplasma species. Thirteen of 18 cats with regenerative anemia were infected, 12 with M. haemofelis and 1 with M. haemominutum. Eight of 22 cats with nonregenerative anemia were infected, 4 with M. haemofelis and 4 with M. haemominutum. Two of 20 cats with normal complete blood (cell) counts were infected with M. haemominutum. Although both mycoplasma species were identified, ill cats were more often infected with M. haemofelis.     

Detection of DNA of 'Candidatus Mycoplasma haemominutum' and Spiroplasma sp. in unfed ticks collected from vegetation in Japan

DNA fragments of 'Candidatus Mycoplasma haemominutum', a feline heamobartonella pathogen, were detected from unfed Ixodes ovatus collected from vegetation in Hokkaido, Fukushima and Yamaguchi Prefectures, and unfed Haemaphysalis flava in Yamaguchi Prefecture. This finding suggests that ixodid tick is a possible vector of 'C. Mycoplasma haemominutum'. Spiroplasma DNA was also detected from unfed I. ovatus in Hokkaido, Fukushima and Yamaguchi Prefectures. The analysis of nucleotides sequence suggested that this Spiroplasma was distinct from registered species.     

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.     

Molecular detection of Mycoplasma wenyonii and 'Candidatus Mycoplasma haemobos' in cattle in Hokkaido, Japan

Blood samples from 78 cattle were tested for hemoplasma infection using molecular methods. PCR and sequence analysis revealed that 17 cattle were infected with Mycoplasma wenyonii, while 13 were infected with 'Candidatus Mycoplasma haemobos'. Four animals were infected with both species. This is the first study to report hemoplasma species infection among cattle in Japan.     

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.     

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.     

Presence of Mycoplasma haemofelis,Mycoplasma haemominutum and piroplasmids in cats from southern Europe: a molecular study

Clinical symptoms produced by Mycoplasma spp. and piroplasmids in cats are sometimes similar. Diagnosis of these pathogens is difficult by microscopic procedures and molecular methods have been used as an alternative. We present in this work, the development of new molecular procedures for diagnosis of the aforementioned organisms, together with a molecular characterization of isolates found in southern European cats.A single PCR-RFLP procedure was designed for diagnosis of Mycoplasma spp. and a seminested PCR-RFLP was designed for diagnosis of piroplasmids. The 16S or 18S rRNA genes of isolates found in clinical samples were partially sequenced in all positive cases.Mycoplasma spp. was detected in 9 (30%) out of 30 symptomatic cats from Spain. Sequencing indicated that 66.6% of these isolates can be ascribed to Mycoplasma haemofelis and only 33.3% to Mycoplasma haemominutum. Partial 16S rRNA sequences obtained in Spanish isolates were very similar to those previously published from the UK and the USA.The presence of piroplasmids (Babesia and Theileria spp.) was studied in 16 cats from Spain (n=13) and Portugal (n=3). Animals analyzed were 10 cats with immunosuppressive viral infection (either FeLV or FIV), 5 asymptomatic cats and 1 cat with Babesia-compatible symptoms. Asymptomatic cats were all PCR-negative. Partial sequencing of 18S rRNA gene demonstrated that the Babesia-symptomatic cat was infected with Babesia canis canis whereas 3 (30%) out of the 10 cats with immunosuppressive viral infection were coinfected with piroplasmids (1 with B. canis canis, 1 with Theileria annae, and 1 with B. canis canis and T. annae both).     

血液直接PCR方法在猪嗜血支原体检测中的应用

为临床上能快速准确检测猪嗜血支原体感染,建立了一种检测猪嗜血支原体的血液直接PCR方法。该方法能特异性地扩增猪嗜血支原体16SrRNA基因中的一段396bp序列,而对猪繁殖与呼吸综合征病毒、猪圆环病毒2型、猪细小病毒、猪伪狂犬病病毒、大肠杆菌、金黄色葡萄球菌等没有扩增到目的条带;该方法能检测到每微升抗凝血中猪嗜血支原体的最低拷贝数为5.52×101 copies,与常规PCR相当;应用血液直接PCR和常规PCR对100份抗凝血样进行检测,阳性检出率分别为65%和69%,差异不显著(P0.05)。结果表明,血液直接PCR方法具有良好的特异性、较高的灵敏度和较快的检出速度,适用于临床上对猪嗜血支原体感染进行快速确诊和流行病学调查。     

Real-time PCR detection of Mycoplasma felis in domestic cats suffering from chronic conjunctivitis (Poland)

Real-time PCR directed to intergenic spacer (IGS) noncoding region between 16S and 23S rRNA genes was used for species specific detection of Mycoplasma felis in conjunctival scrapings. Samples were collected from 57 cats suffering from chronic conjunctivitis in 2008-2010 (Wroc?aw, Poland). Samples from 36 cats (63.2%) were shown to be positive for Mycoplasma felis. Our research gives a first insight in the occurrence of Mycoplasma felis among domestic cats in Poland suggesting that this pathogen may constitute an underestimated cause of chronic conjunctivitis.     

用PCR和探针杂交法检测鸡败血霉形体

根据鸡败血霉形体ｆＭＧ－２核酸片断序列,设计合成了１对２５ｂｐ寡核苷酸引物,对鸡败血霉形体基因组ＤＮＡ进行扩增,均获得预期的７３２ｂｐ扩增产物,检测灵敏度为１ｂｐ;参考菌株ＤＮＡ无扩增。回收纯化琼脂糖电泳凝胶中的扩增产物,ＤＩＧ随机引物法合成核酸探讨,Ｄｏｔ－ｂｌｏｔ杂交试验,鸡败血霉形体呈阳性,检测灵敏度为１００ｐｇ;其他为阴性。对自然发病鸡群检测进一步表明,建立的ＰＣＲ和探针杂交法具有高度的灵     

The use of real-time PCR in the diagnosis and monitoring of Mycoplasma haemofelis copy number in a naturally infected cat

A 5-year-old male neutered cat was diagnosed with severe anaemia due to acute Mycoplasma haemofelis infection. Inflammatory respiratory disease was present concurrently. The cat was treated successfully using a fresh transfusion of whole blood and a 6 week course of doxycycline. The patient made a rapid recovery although the allergic airway disease subsequently required specific therapy consisting of inhaled fluticasone and salbutamol. Real-time quantitative PCR assays confirmed the presence of M. haemofelis DNA copies in the blood at presentation. Repeat PCR assays showed a reduction in copy number during treatment and negative PCR results were obtained both 91 and 425 days after presentation. The report describes, for the first time, the use of real-time PCR in the diagnosis and monitoring of natural M. haemofelis copy number, as well as the induction of long-term negative PCR status.     

Influence of swab material on the detection of Mycoplasma gallisepticum and Mycoplasma synoviae by real-time PCR

Recent reports have shown an increased recovery of cells from flocked nylon swabs which may improve the specimen quality and the real sensitivity of diagnostic tests in a clinical setting. In this study, the detection of Mycoplasma gallisepticum (MG) and M. synoviae (MS), using dry swabs of different materials (nylon flocked, cotton, and polyester), was investigated using real-time TaqMan PCR protocols. Different types of samples, including dilutions of pure broth cultures of MG and MS as well as swabs from tracheas of experimentally infected chickens and field cases of infection, were analyzed. There were no statistical differences in real-time PCR results among the different swab types (P < 0.05), indicating that this is not likely to be a significant factor in MG and MS detection by this method.