In vivo transmission studies of ��Candidatus Mycoplasma turicensis�� in the domestic cat

The natural transmission routes of the three feline haemotropic mycoplasmas – Mycoplasma haemofelis, ‘Candidatus Mycoplasma haemominutum’, and ‘Candidatus Mycoplasma turicensis’ (CMt) – are largely unknown. Since CMt has been detected in the saliva of infected cats using PCR, we hypothesised that direct transmission via social or aggressive contact may occur. The aim of this study was to evaluate this transmission route. CMt-positive saliva and blood samples were obtained from three prednisolone-treated specific pathogen-free (SPF) cats that were infected intraperitoneally with CMt. Five SPF cats were inoculated with CMt-positive saliva or blood subcutaneously to mimic cat bites, and five cats were inoculated orally with blood or oronasally with saliva to mimic social contact. Blood samples were monitored for CMt infection using quantitative real-time PCR and for seroconversion using a novel western blot assay. Neither oronasal nor subcutaneous inoculation with CMt-positive saliva led to CMt infection in the recipient cats, as determined by PCR, independent of prior prednisolone treatment. However, when blood containing the same CMt dose was given subcutaneously, 4 of the 5 cats became PCR-positive, while none of the 5 cats inoculated orally with up to 500 μL of CMt-positive blood became PCR-positive. Subsequently, the latter cats were successfully subcutaneously infected with blood. All 13 CMt-exposed cats seroconverted. In conclusion, CMt transmission by social contact seems less likely than transmission by aggressive interaction. The latter transmission may occur if the recipient cat is exposed to blood from an infected cat.     

The detection of “Candidatus Mycoplasma haemobos” in cattle and buffalo in China

“Candidatus Mycoplasma haemobos” is a hemoplasma species found in cattle and has been recently reported in Switzerland and Japan. In this study, “Candidatus Mycoplasma haemobos” was shown to occur in cattle and buffalo in tropical China by PCR amplification and sequence analysis of the 16S rRNA gene from blood samples. Based on the 16S rDNA sequence, a specific PCR assay was developed. Occurrence of “Candidatus Mycoplasma haemobos” in cattle and buffalo in Guangxi, China, was determined by examining 25 buffalo blood samples, 12 yellow cattle blood samples and 42 dairy cow blood samples. The results showed that 32% (8/25) of buffalo, 41.7% (5/12) of yellow cattle, and 14.3% (6/42) of dairy cows were positive for “Candidatus Mycoplasma haemobos”, respectively. Direct sequencing of representative PCR products confirmed that the amplified partial 16S rDNA sequence represented “Candidatus Mycoplasma haemobos”. This is the first report of “Candidatus Mycoplasma haemobos” in buffalo, yellow cattle, and dairy cows in China.     

Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and “Candidatus Mycoplasma haematoparvum” in dogs

Two canine haemoplasma species have been recognised to date; Mycoplasma haemocanis (Mhc), which has been associated with anaemia in splenectomised or immunocompromised dogs, and “Candidatus Mycoplasma haematoparvum” (CMhp), recently described in an anaemic splenectomised dog undergoing chemotherapy. The study aim was to develop quantitative real-time PCR assays (qPCRs) incorporating an endogenous internal control to detect Mhc and CMhp and to apply these assays to DNA samples extracted from canine blood collected in Northern Tanzania (n = 100) and from dogs presented to a Trinidadian veterinary hospital (n = 185).QPCRs specific for Mhc and CMhp were designed using 16S rRNA gene sequence data, and each was duplexed with an assay specific for canine glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The assays detected ≤10 copies of a sequence-specific haemoplasma plasmid per reaction and neither assay showed cross-reactivity with 10⁶ copies of the sequence-specific plasmid from the non-target canine haemoplasma species.Nineteen of the 100 Tanzanian samples (19%) were positive for Mhc alone and one (1%) was dually infected. One Trinidadian sample was negative for canine GAPDH DNA and was excluded from the study. Of the 184 remaining Trinidadian samples, nine (4.9%) were positive for Mhc alone, five (2.7%) for CMhp alone, and two (1.1%) dually infected.This is the first report of canine haemoplasma qPCR assays that use an internal control to confirm the presence of amplifiable sample DNA, and their application to prevalence studies. Mhc was the most commonly detected canine haemoplasma species.     

Widespread infection with hemotropic mycoplasmas in bats in Spain,including a hemoplasma closely related to “Candidatus Mycoplasma hemohominis”

Molecular analyses of blood samples revealed infection with hemoplasmas in 97% of 31 cave bats captured in three caves in North-Eastern Spain. The characterization of 1250 bp of the 16S rRNA gene in 29 of the positive bats identified two different groups of sequences. Twenty-two Schreibers’ bats (Miniopterus schreibersii) and one long-eared bat (Myotis capaccinii) shared one group, composed of seven closely related sequences. These sequences showed an identity of about 97% with “Candidatus Mycoplasma hemohominis” and the phylogenetic branch including bat and human sequences showed a 100% bootstrap value, supporting a close phylogenetic relationship between these hemoplasmas. The second group, representing a potentially novel species, was composed of a single sequence shared by six Schreibers’ bats that had 91% identity with the recently reported hemoplasma from little brown bats in North America. Large bat aggregations in roosting caves probably benefits intra and inter-species transmission explaining the high observed prevalence.     

The occurrence of the feline “Candidatus Mycoplasma haemominutum” in dog in China confirmed by sequence-based analysis of ribosomal DNA

In the present study, polymerase chain reaction (PCR) assay was used to detect haemoplasmas (haemotropic bacteria) in 40 clinically healthy pet dogs in Foshan city, Guangdong Province, China, and one dog was found positive. Comparison of its 16S ribosomal DNA (rDNA) sequence with relevant sequences showed that the isolated haemoplasma had greater sequence identity to feline species “Candidatus Mycoplasma haemominutum” (99%) than to “Candidatus Mycoplasma haematoparvum” (95%). This result, for the first time, indicates the presence of the feline “Candidatus Mycoplasma haemominutum” in Chinese dogs and it represents the first survey of its kind in China by using PCR assay. The results indicated that dog may represent one of the hosts for the feline “Candidatus Mycoplasma haemominutum”.     

Expansion of intracellular IFN-γ positive lymphocytes during Mycoplasma agalactiae infection in sheep

A method to assess the expansion of antigen-specific intracellular IFN-γ positive T cell subsets during the infection will be helpful for a better understanding of mycoplasmal infections physiopathology in the sheep. We analysed the percentage of antigen-specific lymphocytes positive for intracellular IFN-γ during the infection of sheep with Mycoplasma agalactiae by culturing peripheral blood mononuclear cells of infected or uninfected animals with irradiated M. agalactiae. The expansion of antigen-specific IFN-γ positive lymphocytes in infected sheep was initially sustained by CD4⁺ T cells at day 15 after infection, when antigen specific IgG start to be detectable, followed by CD8/IFN-γ double positive cells. γδ T-cells were not expanded at any time point analysed. IFNγ⁺ T cells disappear 60 days after infection, suggesting that antigen specific IFNγ⁺ T cells, mainly detected in the early phase of the disease, could be useful to understand the role of cell-mediated immunity during M. agalactiae infection.     

Expansion of intracellular IFN-γ positive lymphocytes during Mycoplasma agalactiae infection in sheep

A method to assess the expansion of antigen-specific intracellular IFN-γ positive T cell subsets during the infection will be helpful for a better understanding of mycoplasmal infections physiopathology in the sheep. We analysed the percentage of antigen-specific lymphocytes positive for intracellular IFN-γ during the infection of sheep with Mycoplasma agalactiae by culturing peripheral blood mononuclear cells of infected or uninfected animals with irradiated M. agalactiae. The expansion of antigen-specific IFN-γ positive lymphocytes in infected sheep was initially sustained by CD4⁺ T cells at day 15 after infection, when antigen specific IgG start to be detectable, followed by CD8/IFN-γ double positive cells. γδ T-cells were not expanded at any time point analysed. IFNγ⁺ T cells disappear 60 days after infection, suggesting that antigen specific IFNγ⁺ T cells, mainly detected in the early phase of the disease, could be useful to understand the role of cell-mediated immunity during M. agalactiae infection.     

A retrospective study of serum β-hydroxybutyric acid in 215 ill cats: clinical signs, laboratory findings and diagnoses

Serum concentrations of β-hydroxybutyric acid (sBHBA) are increased in cats with diabetes mellitus (DM), diabetic ketoacidosis (DKA) and hepatic lipidosis (HL). This study assessed sBHBA as a diagnostic tool in 215 consecutively-enrolled ill cats in the general population in a veterinary hospital. At the time of presentation, sBHBA was within the reference range in 158/215 (73.5%) cats (median 0.27; range 0.00-0.49 mmol/L) and elevated in 57/215 (26.5%) cats (median 0.87; range 0.51-21.45 mmol/L). Compared to cats with normal sBHBA, those with increased sBHBA had higher frequencies of anorexia, weight loss, icterus, polyuria/polydipsia, hyperbilirubinaemia, hypertriglyceridaemia, pancreatitis, HL, DM and DKA. They had higher concentrations of bilirubin and triglycerides and lower concentrations of potassium, chloride and total protein. There were positive correlations (P<0.01) between sBHBA and urinary glucose (r=0.42) and ketones (r=0.76), but there were no group differences in dipstick levels of urinary ketones. Cats with DM/DKA and with HL had significantly higher sBHBA compared to other cats. Receiver operator characteristics analysis of sBHBA as a predictor of HL showed that sBHBA was a good predictor of HL. Increased sBHBA occurs frequently in ill cats and provides useful diagnostic information, especially in DM/DKA and HL.     

Case–control risk factor study of methicillin-resistant Staphylococcus pseudintermedius (MRSP) infection in dogs and cats in Germany

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged as a highly drug-resistant small animal veterinary pathogen. Although often isolated from outpatients in veterinary clinics, there is concern that MRSP follows a veterinary-hospital-associated epidemiology. This study's objective was to identify risk factors for MRSP infections in dogs and cats in Germany. Clinical isolates of MRSP cases (n = 150) and methicillin-susceptible S. pseudintermedius (MSSP) controls (n = 133) and their corresponding host signalment and medical data covering the six months prior to staphylococcal isolation were analysed by multivariable logistic regression. The identity of all MRSP isolates was confirmed through demonstration of S. intermedius-group specific nuc and mecA. In the final model, cats (compared to dogs, OR 18.5, 95% CI 1.8–188.0, P = 0.01), animals that had been hospitalised (OR 104.4, 95% CI 21.3–511.6, P < 0.001), or visited veterinary clinics more frequently (>10 visits OR 7.3, 95% CI 1.0–52.6, P = 0.049) and those that had received topical ear medication (OR 5.1, 95% CI 1.8–14.9, P = 0.003) or glucocorticoids (OR 22.5, 95% CI 7.0–72.6, P < 0.001) were at higher risk of MRSP infection, whereas S. pseudintermedius isolates from ears were more likely to belong to the MSSP-group (OR 0.09, 95% CI 0.03–0.34, P < 0.001). These results indicate an association of MRSP infection with veterinary clinic/hospital settings and possibly with chronic skin disease. There was an unexpected lack of association between MRSP and antimicrobial therapy; this requires further investigation but may indicate that MRSP is well adapted to canine skin with little need for selective pressure.     

Florfenicol feed supplemented decrease the clinical effects of Mycoplasma hyopneumoniae experimental infection in swine in México

The therapeutic value of Florfenicol feed supplemented was evaluated in conventional pigs to eliminate consequences of chronic infection of Mycoplasma hyopneumoniae. The experimental animals were pigs with an average of 16 kg, after intratracheally inoculation with M. hyopneumoniae they were divided in two experimental groups: (a) the non-medicated; and (b) the feed supplemented group (20 g Florfenicol/ton of feed) during the ensuing 35 days. The average daily weight gain of the Florfenicol-treated pigs (0.33±0.14 kg/day) was significantly higher than that of the non-treated ones (0.21±0.10 kg/day). In medicated animals was still impaired relative to that of the uninfected ones control group (0.39±0.02 kg/day). The average percentage of pneumonic gross lesions extensions' of the pigs groups was: 13.99% for M. hyopneumoniae infected non-medicated group; 1.79% M. hyopneumoniae infected, Florfenicol-treated group and, 0.56% of the uninfected control group. M. hyopneumoniae; colonization was detected at these levels in 7 and 9 members of the respective infected groups. The extent of the pneumonic lesions and M. hyopneumoniae generally was greater in the non-medicated pigs. Therefore, oral administration of Florfenicol via feed ingestion seemed to be somewhat effective in ameliorating the clinical effects of M. hyopneumoniae infection of swine.     

Comparison of the TONOVET Plus®, TonoVet®, and Tono-Pen Vet™ tonometers in normal cats and cats with glaucoma

Objectives

To determine the accuracy, precision, and clinical applicability of the ICare® TONOVET Plus (TVP) in cats.

Animals and Procedures

IOP readings obtained with the TVP were compared to values obtained concurrently with the original TONOVET (TV01) and Tono-Pen Vet™ (TP) in 12 normal cats (24 eyes) and 8 glaucomatous LTBP2-mutant cats (13 eyes) in vivo. Reproducibility of TVP readings was also assessed for three observers in the above cats. The anterior chambers of five different normal cat eyes were cannulated ex vivo. IOP was measured with the TVP, TV01, and TP at manometric IOPs ranging from 5 to 70 mmHg. Data were analyzed by linear regression, ANOVA and Bland–Altman plots. ANOVA was used to assess reproducibility of TVP readings obtained by different observers and an ANCOVA model controlled for variation of individual cats. p < .05 was considered significant.

Results

TVP values strongly correlated with TV01 values (y = 1.045x + 1.443, R² = .9667). The TP significantly underestimated IOP relative to the TVP and TV01, particularly at high IOP. IOP values obtained by 1 observer were significantly higher (~1 mmHg average) compared to the other 2 observers via ANCOVA analysis (p = .0006479 and p = .0203). Relative to manometry, the TVP and TV01 were significantly more accurate (p < .0001) and precise (p < .0070) than the TP in ex vivo eyes.

Conclusions

IOP readings obtained with the TVP and TV01 are broadly interchangeable between models and between observers, but subtle differences may be important in a research context. TP readings vastly underestimate high IOP in feline glaucoma.     

Ollulanus tricuspis in domestic cats — prevalence and methods of post-mortem diagnosis

Ollulanus tricuspus is a parasite of the stomach of domestic cats and other animals, with a worldwide distribution. It can sometimes cause severe gastritis.

Fifty-five cat stomachs were examined for O.tricuspis using three techniques. O.tricuspis was found in seven stomachs 13%) from adult cats; the number of parasites recovered from individual cats ranged from 9 to 119 (mean 54). The prevalence was higher than that previously recorded in New Zealand. Repeated dilution and sedimentation of the stomach contents and mucosal washings was found to be the most reliable detection method, being positive in all of the detected infections. Pepsin/HCI digestion of the stomach mucosa detected only 71% and punch sampling of the mucosa only 29% of these infections. This contrasts with an overseas study indicating that, in heavily infected cats, these techniques are similar in sensitivity. No macroscopic lesions were seen in the stomachs of infected cats.     

Can heartworm prevalence in dogs be used as provisional data for assessing the prevalence of the infection in cats?

Cats are considered a susceptible host for Dirofilaria immitis; however, increased host resistance is reflected by relatively low adult worm burdens in natural and experimental infections; the prolonged prepatent period (8 months); the low level and short duration of microfilaremia; and the short life span of adult worms (2-3 years). From April to September 2006, 212 cats and 608 dogs, all exposed for at least one transmission season, were screened for D. immitis infection in a multi-center study in the Po River Valley in northern Italy. Cats were initially evaluated by antibody testing; positive subjects were followed up by antigen testing and echocardiography (and necropsy if death occurred). The prevalence in dogs was 29% by a modified Knott test and antigen testing compared with a prevalence of 4.7% in cats by an antibody test; six of these infections (2.8%) were confirmed by the follow-up evaluations. This field study demonstrated that the prevalence of heartworm infection in cats in this area is within the expected limits of 9-18% of the prevalence in dogs. Antibody testing likely underestimates the real prevalence of D. immitis infection in cats. These results also emphasize the importance of preventive treatment in cats.     

Tick paralysis in dogs and cats in Australia: treatment and prevention deliverables from 100 years of research

This review of tick paralysis caused by Ixodes holocyclus in Australia addresses the question: What are the key discoveries that have enabled effective treatment and prevention of tick paralysis in dogs and cats? Critical examination of 100 years of literature reveals that arguably only three achievements have advanced treatment and prevention of tick paralysis in animals. First, the most significant treatment advance was the commercial availability of tick antiserum in the 1930s. Hyperimmune serum currently remains the only specific anti-paralysis tick therapy available to veterinarians in Australia. Second, advances in veterinary critical care have increased survival rates of the most severely affected dogs and cats. Critical care advancements have been enabled through specialised veterinary hospitals that can provide appropriate care 24 h a day, and advanced training of veterinarians, veterinary nurses and technicians. Third, perhaps that biggest advance of all in the last 100 years of research has been the commercial availability of the isooxazoline class of acaricidal preventatives in Australia specifically for I. holocyclus. This highly effective class of preventatives offers long duration of action, low cost, spot-on or oral formulations and a low rate of adverse reactions. Animal owners and veterinarians now have the most useful tool of all – a reliable preventative. This review reveals the key events in research over the last 100 years and the tortuous pathway to delivering better treatment and preventative options for this enigmatic Australian parasite.     

Anemia in cats with hemotropic mycoplasma infection: Retrospective evaluation of 23 cases (1996–2005)

This study summarizes the diagnostic findings from all anemic cats diagnosed with hemotropic mycoplasma (HM) infections at the Western College of Veterinary Medicine — Veterinary Teaching Hospital between 1996 and 2005. The objectives were to determine the frequency of HM-induced anemia among all cats presented with anemia during this period, the clinical findings and risk factors associated with clinical HM infection, and factors affecting or predicting survival. Medical records were examined from 23 cats with HM-induced anemia from the total of 170 cats diagnosed with anemia during this period. The frequency of HM-induced anemia was 14% (23/170) among all anemic cats. Cats with HM-induced anemia were less likely to be purebred (P = 0.04) than other cats with anemia. Of the cats with HM-induced anemia, those with positive retroviral status (P = 0.01), concurrent illness (P < 0.01), or lack of erythroid regeneration (P = 0.01) were most likely to die. The 1-year survival of HM-infected cats was 65% (13/20).     

Intestinal parasites in dogs and cats from the district of Évora, Portugal

Intestinal parasites, both helminths and protozoa, are commonly found in domestic animals, and the possible transmission of enteric parasites from dogs and cats to humans may constitute a global potential health risk worldwide. In the present study, we analysed 148 stool samples from dogs (n=126) and cats (n=22) collected from animal shelters and veterinary clinics, in the district of évora, Portugal. Microscopic examination confirmed that Giardia was the most frequent parasite in the studied population (34/148; 23%). Other parasites such as Ancylostoma sp., Isospora spp., Toxocara, Trichuris spp., Toxascaris and Toxoplasma were also found. Furthermore, molecular characterization of Giardia duodenalis analysis targeting the small subunit ribosomal RNA (ssu-rRNA) was performed revealing the presence of host-specific (C and D) and zoonotic assemblages (A and B). This work points out to the importance of protozoan parasites in companion animals, and reanalyses the need for parasite prophylaxis.     

Measurement of pulmonary transit time in healthy cats by use of ultrasound contrast media “Sonovue®”: Feasibility,reproducibility, and values in 42 cats

ObjectiveTo evaluate the feasibility of measuring pulmonary transit time (PTT) in healthy cats by transthoracic echocardiography using the ultrasound contrast agent Sonovue^®. To determine normalized PTT (nPTT) values in 42 healthy cats and to estimate the interobserver variability and the within-day repeatability of nPTT measurements.AnimalsForty-two privately owned healthy cats of different breeds, gender and age presented for cardiac examination.MethodsA bolus injection of contrast agent (Sonovue^®) was administered intravenously. The right parasternal short axis echocardiographic view was used to record the contrast agent's transit time from the pulmonary artery to the left atrium. Pulmonary transit time and nPTT were determined independently by three examiners with different levels of experience.ResultsNormalized PTT was 4.12 ± 1.0 (mean ± SD) in our population. The median interobserver variability across our population was 6.8%, the median within-day variability for the three observers were 13.1%, 12.7% and 13%. No effect of the observer's experience on nPTT measurement was identified. Age, sex and body weight did not significantly influence nPTT.ConclusionsThis study demonstrates that nPTT measurement is feasible in cats using ultrasound and the blood pool contrast media Sonovue^®. Measurements of nPTT can be performed in a clinical setting. Normalized PTT values in healthy cats are comparable with those reported in healthy dogs.     

Successful balloon angioplasty of pulmonary artery stenosis in two cats and associated complications

Two cats (2.5 months and 8 months old) were each evaluated due to a loud systolic murmur, and each was diagnosed with severe pulmonary artery stenosis at the bifurcation of the main pulmonary artery. Echocardiograms confirmed significant right atrial dilation and right ventricular dilation and/or hypertrophy that was progressive in one cat. Atenolol was initiated and the cats were referred for interventional therapy. Balloon angioplasty was performed via the jugular vein. In case 1, the pressure gradient across the stenosis was reduced from 169.7 mmHg to 23.6 mmHg and 52.4 mmHg across the left and right branch pulmonary arteries, respectively. In case 2, the stenotic echocardiographic gradient was reduced from 64 mmHg to 38.0 mmHg and 35.3 mmHg across the left branch and right-branched pulmonary arteries respectively. Both patients developed moderate to severe dynamic right ventricular outflow tract obstruction post angioplasty. Case 2 developed hypotension, desaturation, and ventricular arrhythmias intra-operatively. Case 1 was discharged but appeared to develop acute lung perfusion injury approximately 36 h after procedure that was manifested by radiographic pulmonary congestion and pulmonary infiltrate of the left lung fields. The congestion was successfully managed medically. Serial echocardiograms over the following 4 years in case 1 showed near complete resolution of the stenosis and associated right heart enlargement.