Mycoplasma as a cause of canine urinary tract infection

Mycoplasmas were isolated from 60 specimens of urine obtained by cystocentesis from 41 dogs (23 males and 18 females) with urinary tract infection. Mycoplasmas were isolated in pure culture from 41 (68%) of the specimens, and were isolated in conjunction with one or more bacterial species from 19 (32%) specimens. Clinical signs of urinary tract infection were noted in 20 of 31 dogs in which mycoplasmas were isolated in pure culture, and numbers of WBC in the urine sediment were above the reported normal range in 22 of 25 urine specimens from those 20 dogs. Twenty-four of 29 mycoplasma isolates were found to be Mycoplasma canis, 4 were found to be M spumans, and 1 was identified as M cynos.     

Mycoplasma canis and urogenital disease in dogs in Norway

Mycoplasmas identified as Mycoplasma canis were isolated from nine dogs with clinical signs of urogenital disease in Norway over a period of 20 months. Some of the dogs had been treated unsuccessfully with antibiotics, and three were euthanased as a result of severe persistent disease. Seven of the dogs had a urinary tract infection, one had chronic purulent epididymitis and one had chronic prostatitis. Overt haematuria was frequently observed among the dogs with cystitis. M canis was isolated in pure culture from seven of the dogs and in mixed culture from the other two. In three cases the mycoplasma was cultivated only from urinary sediment, and it was typically obtained in smaller numbers than would be considered indicative of a urinary tract infection. In contrast with most mycoplasmas, the M canis isolated from all the dogs grew on ordinary blood agar plates used for routine bacteriological cultivation. Specific mycoplasma media were not used and the presence of other Mycoplasma or Ureaplasma species cannot be excluded.     

Molecular characterization of Babesia canis canis isolates from naturally infected dogs in Poland

Babesia canis has generally been considered the only large Babesia to infect dogs. In this study, we used PCR to detect and characterize B. canis canis isolated from naturally infected dogs in Poland by amplifying and sequencing a portion of the 18S ribosomal RNA (rRNA) gene. Venous blood samples were collected from 76 Babesia-symptomatic dogs. A 559-bp fragment of the B. canis canis 18S rRNA gene was amplified by PCR. The PCR products were then digested with HincII restriction enzyme, and isolates were classified according to whether they were cut (group A) or not (group B) by this endonuclease. Sequencing of the PCR products from the isolates led to the identification of seven sequence variants (four in group A, and three in group B). Sequences were compared with GenBank sequences, and alignments showed that all B. canis canis isolates from Europe may be classified into groups A or B as defined in our study.     

Canine mycoplasmas

This review aims to summarise our current understanding of the role of mycoplasmas in domestic dogs. Canine mycoplasmology is a small field, with less than 50 publications in the past 40 years. In this time we have gained knowledge about the number of species and have made associations with infections in dogs. However much evidence is still lacking. The importance of all canine mycoplasmas remains unknown, yet certain species are associated with canine anaemia (Mycoplasma haemocanis), respiratory disease (Mycoplasma cynos) and urogenital tract infections (Mycoplasma canis). Mycoplasmas can be isolated in pure culture from canine clinical specimens and it is hoped that this review will stimulate veterinarians to consider mycoplasmas as a potential cause of disease in dogs, especially when antibiotic therapy is failing.     

Mycoplasma species and related organisms isolated from ruminants in Britain between 1990 and 2000

Between 1990 and 2000, more than 1600 mycoplasmas and the related acholeplasmas were identified from ruminant animals by the Mycoplasma Group at the Veterinary Laboratories Agency--Weybridge. Mycoplasma bovis was the most commonly identified pathogen, mostly from pneumonic calves but occasionally from cattle with mastitis and arthritis. Mycoplasma canis was first isolated in Britain in 1995 from pneumonic calves and the number of isolates increased to 18 per cent of the total mycoplasmas isolated from cattle in 1999. The ELISA for antibodies to M. bovis detected 1971 positive samples (22 per cent) among 8959 serum samples, mainly from pneumonic cattle. Other mycoplasmas identified included Mycoplasma dispar from the lungs of cattle with respiratory disease, and Mycoplasma bovigenitalium from the reproductive tract of cows with vulvovaginitis and infertility. Mycoplasma bovirhinis and Acholeplasma species were found commonly but are thought to be more opportunistic than pathogenic. In sheep and goats, the majority of Mycoplasma species isolated were identified as Mycoplasma ovipneumoniae from pneumonic sheep, Mycoplasma conjunctivae from sheep with keratoconjunctivitis, and the ubiquitous Mycoplasma arginini.     

New advances in molecular epizootiology of canine hematic protozoa from Venezuela, Thailand and Spain

The prevalence of hematozoan infections (Hepatozoon canis and Babesia sp., particularly Babesia canis vogeli) in canids from Venezuela, Thailand and Spain was studied by amplification and sequencing of the 18S rRNA gene. H. canis infections caused simultaneously by two different isolates were confirmed by RFLP analysis in samples from all the geographic regions studied. In Venezuela, blood samples from 134 dogs were surveyed. Babesia infections were found in 2.24% of the dogs. Comparison of sequences of the 18S rRNA gene indicated that protozoan isolates were genetically identical to B. canis vogeli from Japan and Brazil. H. canis infected 44.77 per cent of the dogs. A representative sample of Venezuelan H. canis isolates (21.6% of PCR-positives) was sequenced. Many of them showed 18S rRNA gene sequences identical to H. canis Spain 2, albeit two less frequent genotypes were found in the sample studied. In Thailand, 20 dogs were analyzed. No infections caused by Babesia were diagnosed, whereas 30 per cent of the dogs were positive to hematozoan infection. Two protozoa isolates showing 99.7-100% identity to H. canis Spain 2 were found. In Spain, 250 dogs were studied. B. canis vogeli infected 0.01% of the animals. The sequence of the 18S rRNA gene in Spanish isolates of this protozoa was closely related to those previously deposited in GenBank (> 99% identity). Finally, 20 red foxes were screened for hematozoans employing semi-nested PCR and primers designed to detect Babesia/Theileria. Fifty percent of the foxes were positive to Theileria annae. In addition, it was found that the PCR assay was able as well to detect Hepatozoon infections. Thirty five percent of the foxes were infected with two different H. canis isolates showing 99.8-100% identity to Curupira 1 from Brazil.     

Detection of antibodies to mycoplasmas using an immunoenzyme method

Detection of the antibodies to the species Mycoplasma bovis in the serum and milk of dairy cows coming from a mastitis-infected herd is a good example of utilization of the ELISA immunoenzymologic method in the mycoplasmology. Examining the samples from 75 dairy cows and applying the indirect hemagglutination test, good correlation of the results of the two tests was determined. The antibodies to the species Ureaplasma diversum were demonstrated by the ELISA method both in the bovine serum and in the milk of dairy cows infected slightly with mastitis. We chosen that strain which detected the maximum titres in the selected samples of the sera out of four antigens prepared from various strains of U. diversum. Rabbit sera hyperimmune to 26 strains of the mycoplasmas of various species were used to identify two antigens (after removing the antibodies to the components of the media). Specific reaction was obtained with the antisera to M. hyorhinis and M. arginini.     

Molecular studies on Babesia,Theileria and Hepatozoon in southern Europe. Part I. Epizootiological aspects

Molecular epizootiology of piroplasmids (Babesia spp., Theileria spp.) and Hepatozoon canis was studied in mammals from southern Europe (mainly from Spain, but also from Portugal and France). Partial amplification and sequencing of the 18s rRNA gene was used for molecular diagnosis. In some particular cases (B. ovis and B. bovis) the complete 18s rRNA gene was sequenced. Blood samples were taken from domestic animals showing clinical symptoms: 10 dogs, 10 horses, 10 cows, 9 sheep and 1 goat. In addition, DNA samples were isolated from blood of 12 healthy dogs and from spleen of 10 wild red foxes (Vulpes vulpes). The results of the survey were the following: Piroplasmid infections: Approximately from 50 to 70% of wild or domestic mammals (symptomatic) were infected.Piroplasmids detected in ruminants were:COW: B. bovis, T. annulata and Theileria sp. (type C). Sheep and goat: B. ovis. Piroplasmids present in canids were: Babesia canis vogeli, Babesia canis canis, Theileria annae and B. equi. The only piroplasmid found in asymptomatic dogs was B. equi. Piroplasmids found in horse were: B. equi and B. canis canis.H. canis infections in canids: H. canis was absent of domestic dog samples, whereas all foxes studied were infected by this protozoa.Genetic analysis showed that most of piroplasmid and Hepatozoon isolates from southern Europe matched unambigously with previously described species, as demonstrated by the high level sequence identity between them, usually between 99 and 100%. Minor differences, usually detected in hypervariable regions of 18s rRNA gene are probably due to strain variations or rare genetic polymorphisms. A possible exception was B. bovis, which shows a relatively lower degree of homology (94%) with regard to other B. bovis isolates from several countries. The same is true for B. ovis, that showed a 94% identity with regard to Babesia sp. from South African cow and a 92% with rapport to B. bovis from Portugal.     

Toxocara eggs shed by dogs and cats and their molecular and morphometric species-specific identification: is the finding of T. cati eggs shed by dogs of epidemiological relevance?

Intestinal infections with Toxocara cati and Toxocara canis in their definitive host (felids and canids, respectively) are diagnosed by egg identification in faeces using coproscopical techniques. The Toxocara species is assumed to comply with the species from which the examined faeces were obtained, i.e. T. cati in cats and T. canis in dogs. We isolated and measured Toxocara eggs from faecal samples of 36 cats and 35 dogs from Switzerland and identified the Toxocara species by PCR. Amongst the isolates originating from dogs, 24 (68.5%) were determined as T. canis and 11 (31.5%) as T. cati. In all samples originating from cats, only T. cati was identified. Based on PCR identification, eggs of T. canis (n=241) and T. cati (n=442) were measured, revealing statistically significant different (p<0.001) mean sizes of 62.3 by 72.7 μm for T. cati and 74.8 by 86.0 μm for T. canis eggs. Considering that coprophagy is not unusual for dogs, a considerable percentage of Toxocara infections coproscopically diagnosed in dogs, as well as assumptions on anthelminthic resistance in regularly treated dogs, might in fact relate to intestinal passages of eggs following the uptake of other animals' faeces.     

Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50

The surface antigen P50 of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding truncated P50 (P50t) lacking a signal peptide and C-terminal hydrophobic regions were cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). More than 90% portion of the GST-P50t was expressed as a soluble form, in contrast with GST-P50f (full-length), which was completely expressed as an insoluble form. This result indicates that removal of the hydrophobic signal peptide and C-terminus had dramatically improved its hydrophilicity. The purified GST-P50t was tested in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to B. gibsoni in dogs. The ELISA with GST-P50t clearly differentiated between B. gibsoni-infected dog sera and uninfected dog sera. In addition, the ELISA detected no cross-reactivity with sera from dogs experimentally infected with the closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi. Field serum samples collected from dogs in Japan and China were examined for the diagnosis of B. gibsoni infection by using the ELISA. 14.5% (9/62), 5.8% (7/120), and 5.4% (2/37) of tested samples were positive for dogs from Okinawa, Yamaguchi, and Osaka prefectures, Japan, respectively. On the other hand, 4.8% (2/41) of tested samples were positive for dogs from Nanjing, China. These results suggest that the GST-P50t could be a reliable reagent for practical use in ELISA for the serodiagnosis of canine piroplasmosis caused by B. gibsoni.     

Dermatophytes in clinically healthy dogs and cats

The hair and skin of 300 clinically healthy animals, 268 dogs and 32 cats, were examined mycologically. The method described by Mariat and Tapia (1966) was used for the examination, along with square pieces of the fitted carpet Kovral. The dermatophytes were isolated in 12 samples, all from the material taken from dogs. Trichophyton mentagrophytes was isolated six times, Microsporum canis four times, Trichophyton gallinae once, Trichophyton rubrum once. The origin of the dermatophyte Trichophyton gallinae, found in a village dog, was not determined. In one case of occurrence of the dermatophyte Trichophyton mentagrophytes in a dog the dermatophyte was probably transmitted from man to the dog. Microsporum canis was not proved to be the most frequent dermatophyte in dogs or cats in this country.     

Studies of bovine Mycoplasma mastitis. 4. Serological classification of Mycoplasma strains isolated from 3 stocks

Thirty-two isolates from altered milk samples taken from cows with mastitis of two herds were serologically tested by the WHT against rabbit hyperimmune sera of 16 reference and type strains. The strains tested were associated and grouped in the following way: 16 M. bovis, nine A. laidlawii, and seven A. axanthum. The classification of on M. bovis strain was confirmed by SHT. Five isolates from another stock, with biochemical properties related to the family of mycoplasmataceae, were not serologically identified.     

Antibodies to Ehrlichia canis, Ehrlichia platys, and Spotted Fever Group Rickettsiae in Louisiana Dogs

Antibodies to Ehrlichia canis, Ehrlichia platys, and spotted fever group (SFG) rickettsiae were detected by indirect immunofluorescence in sera from 27 ill individually owned thrombocytopenic dogs (platelet concentrations less than 200,000 platelets/microliters) and 59 healthy kenneled dogs located in southern Louisiana. Platelet concentrations less than 100,000 platelets/microliters were detected in 63% of ill thrombocytopenic dogs and 6.8% of healthy kennel dogs. One ill thrombocytopenic dog had intracytoplasmic E platys morulae detected within platelets. The prevalence of increased serum antibody titers to E canis and E platys was 25.9% and 40.7% for the ill thrombocytopenic dogs and 20.3% and 54.2% for the healthy kennel dogs, respectively. All dogs with seropositivity to E canis had increased antibody titers of greater than or equal to 1:100 to E platys. Simultaneous examination of increased serum antibody titers (greater than or equal to 1:64) to four SFG rickettsiae indicate that Rickettsia rhipicephali and Rickettsia montana accounted for the majority of the antibodies detected in these dogs. Of 86 dogs tested, 44.2% were seronegative to E canis, E platys, and SFG rickettsiae.     

Cultivation of mycoplasma from conjunctiva and production of corneal immune response in guinea pigs.

Mycoplasma canis was the most frequent mycoplasmal isolant obtained from the conjunctival surface of the eyes of dogs. Nine mycoplasmal isolates, 5 were M canis, were recovered from the lower cul-de-sac of 101 dogs. Experimentally induced immune keratitis was produced by sensitizing guinea pigs to 3 antigen preparations of M canis and then administering intracorneal challenge inoculation. The guinea pigs were clinically observed by leukocyte migration-inhibition (LMI) technique and by histopathologic study of their corneas. The corneal reaction of the guinea pigs was not identical to a clinical entity of the dog.     

Isolation of Microsporum canis from the hair coat of pet dogs and cats belonging to owners diagnosed with M. canis tinea corporis

Microsporum canis has been frequently isolated from human cases of tinea capitis and tinea corporis. The infection may be acquired from infected animals with cutaneous lesions but also from asymptomatic carriers or from the environment. As asymptomatic M. canis carriers are considered to be a critical factor in the epidemiology of dermatophytosis in humans, this study investigated the relationship between the presence of dermatophytes on the hair coats of dogs and cats without cutaneous lesions and the occurrence of the disease in their respective owners. A total of 136 dogs and 248 cats were sampled from January 1999 to January 2005. Seventy-eight animals (22 dogs and 56 cats) belonged to individuals affected by tinea corporis caused by M. canis and 306 (114 dogs and 192 cats) to individuals without dermatophytosis. Age, sex, breed, habitat and season were recorded for each animal and examined as potential risk factors. Dermatophytes were isolated from 20.5% of the dogs and 28.2% of the cats. Microsporum canis was isolated from 36.4% of dogs cohabiting with owners diagnosed with tinea corporis but it was never isolated from dogs whose owners had no lesions. By contrast, M. canis was isolated from 53.6% of cats cohabiting with owners diagnosed with tinea corporis and from 14.6% of cats whose owners had no signs of the disease. These results clearly indicate that both cats and dogs should be considered as a major source of pathogenic dermatophytes for humans even when they do not present clinical signs of dermatophytosis.     

Serological survey for Ehrlichia canis in urban dogs from the major population centres of northern Australia

OBJECTIVE: To detect evidence of Ehrlichia canis infection of dogs from the major population centres of northern Australia, if present. DESIGN: Serological investigation for E. canis. PROCEDURE: The sera of 316 domestic dogs, collected from the northern Australian population centres of Townsville, Cairns, Darwin, Kununurra and Broome from May 1997 to August 1999, were investigated for evidence of infection with E. canis. Samples were tested for antibodies to E. canis using an indirect fluorescent antibody (IFA) test. The buffy coats from blood of dogs whose serum reacted in the IFA test were subsequently tested with a nested PCR to detect E. canis DNA. When available, blood from these dogs was injected into suckling mice, which were then examined for clinical disease and tested for the presence of E. canis antibodies. RESULTS: Of the 316 samples tested seven reacted in the IFA test for E. canis. None of the dogs from which these samples were obtained exhibited clinical signs of acute or chronic ehrlichiosis. The six positive samples available for testing were negative when tested with the nested PCR. Suckling mice inoculated with blood from three of the dogs whose serum was positive by IFA test showed no signs of clinical disease nor did their give positive reactions in the IFA test. CONCLUSIONS: No evidence of E. canis infection was confirmed in any of the dogs examined. Northern Australia would appear to remain free of this obligate parasite.     

Isolation of mycoplasma species from the lower respiratory tract of healthy cattle and cattle with respiratory disease in Belgium

Between 1997 and 2000, a total of 150 healthy cattle and 238 animals with respiratory disease were examined for six Mycoplasma species. Attempts were made to detect Mycoplasma canis, Mycoplasma dispar and Ureaplasma diversum in calves with recurrent disease, and all three of these species were identified in calves with recurrent disease and in healthy lungs. In healthy calves, 84 per cent of bronchoalveolar lavage fluids were mycoplasma free; when cultures were positive, Mycoplasma bovirhinis was the only species isolated. Mycoplasmas were isolated from 78 per cent of animals suffering recurrent respiratory disease and from 65 per cent of acute respiratory cases. Mycoplasma bovis was isolated from bronchoalveolar lavages from 35 per cent of calves suffering recurrent respiratory disease, and from 50 per cent of acute cases, and from 20 per cent of pneumonic cases examined postmortem. M bovis was associated with other Mycoplasma species in 44 per cent of cases. M dispar was also isolated from 45.5 per cent of calves suffering recurrent respiratory disease, often in association with M bovis. M canis was identified for the first time in diseased Belgian cattle. Other mycoplasmas, including Mycoplasma arginini, Mycoplasma alkalescens and U diversum, were isolated less frequently. Associations between mycoplasmas and other pathogens were often observed. Among lungs infected with Pasteurella and/or Mannheimia species, more than 50 per cent were mixed infections with M bovis.     

Frequency of yeasts and dermatophytes from healthy and diseased dogs.

The aim of this study was to investigate the presence of dermatophytes and yeasts in healthy and diseased dogs. A total of 633 samples were collected from 26 healthy animals (104 samples), 131 with dermatitis (343 samples), 74 with otitis (148 samples), and 19 with ocular diseases (38 samples). Cultures from healthy animals were positive for Malassezia pachydermatis in 13.5% (7/52) of samples from skin, 42.3% (11/26) from ear, and 3.8% (1/26) from eye. Fungal growth was observed in 20.4% (70/343) samples from animals with dermatitis. Microsporum canis was the most isolated fungus (n = 39), followed by M. pachydermatis (n = 30) and Malassezia sp. (n = 3). Of the 148 samples from dogs with otitis, 90 (60.8%) were positive for M. pachydermatis, and of the clinical specimens from the conjunctiva of animals with ophthalmic disease, 2.6% (1/38) presented positive cultures for M. pachydermatis. Only 14.3% (2/14) of the positive cultures for M. pachydermatis and 40.9% (9/22) of those for M. canis were positive in the direct exam. Direct exams were positive in 84.3% (70/83) of the culture positive samples from affected ears of dogs with otitis. Malassezia pachydermatis may act as an aggravating factor in the occurrence of cutaneous diseases, or the isolation of M. canis may be associated with the onset of dermatophytosis. Fungal culture, rather than microscopic examination, should be used as the definitive diagnostic test for dermatomycoses and otitis.     

Generalized Microsporum canis dermatophytosis in six Yorkshire terrier dogs

Six Yorkshire terrier dogs with generalized, chronic dermatophytosis caused by Microsporum canis were seen over a 3-year period. Specific tests showed that they also had concurrent leishmaniosis (four cases), leishmaniosis and ehrlichiosis (one case) or diabetes mellitus (one case). Although specific therapy for these infectious diseases was instituted and the dogs were treated systemically and topically with appropriate antifungal drugs, only partial clinical resolution of the dermatophytosis was achieved. M. canis infection resolved in the dog with diabetes mellitus after stabilizing the diabetes mellitus. Although immunological studies were not performed in these cases, it is theorized that the immune disregulation caused by leishmaniosis, ehrlichiosis or diabetes mellitus may have favoured generalization of the infection and prevented favourable responses to appropriate treatment of the M. canis infection.     

Hair coat contamination with zoonotic helminth eggs of farm and pet dogs and foxes

Infections of dogs with Toxocara canis and Echinococcus multilocularis pose an infection-risk particularly for contact persons. We examined specimens of hair coat and faeces of 124 farm dogs, 118 household dogs, 49 kennel dogs, 15 puppies from two litters, and 46 red foxes. Microscopically identified eggs of Toxocara or taeniids were further investigated by species-specific PCRs. In farm dogs, eggs of E. multilocularis or T. canis were identified in each 2.4% of faecal samples, eggs of T. cati (gastrointestinal passage) in 7.3%, respectively. Household dogs excreted eggs of T. canis (0.8%) and of T. cati (2.5%). In kennel dogs, eggs of T. canis (4.1%), but not of T. cati were detectable. Coat samples contaminated with eggs of Toxocara spp. were found from farm dogs (5.6%), household dogs (1.7%) and kennel dogs (2.0%). Taeniid eggs were isolated from the coat samples from only two farm dogs (1.6%); a molecular species determination was not achieved. In six intrauterinely infected puppies, Toxocara-eggs were found in 17/38 samples taken within six weeks. No intact Toxocara eggs could be isolated from the coat of nine puppies from a second litter 13 days after deworming. Of the 46 red foxes investigated (dissection and faecal samples) 13 (28.3%) were infected with E. multilocularis and 20 (43.5%) with Toxocara. Eggs of taeniids and Toxocara were found in 13% (in three cases confirmed as E. multilocularis) and 21.7%, respectively, of the coat samples. None of the retrieved Toxocara eggs in the coat samples were embryonated. Thus, an infection of humans through the transmission of E. multilocularis eggs after direct contact with dogs or foxes is conceivable, whereas a corresponding infection risk by Toxocara eggs must be critically challenged.