Clinical presentation, diagnostic investigations, treatment protocols and outcomes of dogs diagnosed with tick-borne diseases living in the United Kingdom: 76 cases (2005-2019).

OBJECTIVES: To report the presence of tick-borne diseases in dogs living in the United Kingdom. MATERIALS AND METHODS: Dogs with a final diagnosis of tick-borne diseases made between January 2005 and August 2019 at seven referral institutions in the United Kingdom were included in the study. RESULTS: Seventy-six dogs were included: 25 were diagnosed with ehrlichiosis, 23 with babesiosis, eight with Lyme borreliosis and six with anaplasmosis. Fourteen dogs had co-infections with two or three pathogens. Except for those dogs with anaplasmosis and Lyme borreliosis, most dogs with tick-borne diseases had a history of travel to or from endemic countries. However, three dogs with ehrlichiosis, and one dog each infected with Babesia canis and Babesia vulpes did not have any history of travel. A variety of non-specific clinical signs and laboratory abnormalities were reported. Targeted treatment was successful at achieving clinical remission in 64 (84%) dogs. CLINICAL SIGNIFICANCE: Even in non-endemic areas, veterinary surgeons should consider tick-borne diseases in dogs with compatible clinical presentation and laboratory findings and especially where there is a history of travel. As autochthonous transmission of tick-borne-pathogens does occur, an absence of travel should not rule out tick-borne diseases. Specific diagnostic testing is required to confirm infection, and this enables prompt targeted treatment and often a positive outcome.

Chronic diarrhea secondary to canine leishmaniosis: Case series.

Chronic diarrhea is a clinical sign associated with canine leishmaniosis, varying from 3 % to 30 % of prevalence. However, its occurrence in dogs has been mostly associated with chronic kidney or liver disease. Leishmania organisms can cause inflammation of the digestive tract with chronic diarrhea as the only clinical manifestation, although it has been poorly documented in dogs. The aim of this retrospective observational study was to describe dogs with chronic diarrhea as the main clinical sign associated with leishmaniosis. All cases had a complete blood count, biochemistry, urinalyses, and diagnostic tests for leishmaniosis. Exclusion criteria included renal or hepatic disease and/or previous diagnosis of gastrointestinal disease. Twenty-three dogs were included. Small bowel diarrhea was present in 7/23 (30.4 %), large bowel diarrhea in 9/23 (39.2 %) and mixed diarrhea in 7/23 (30.4 %). Gastrointestinal biopsies were performed in 8/23 dogs and Leishmania amastigotes were found in all of them. In the others, leishmaniosis was diagnosed by serology in 10/15 dogs (66.7 %), serology plus blood PCR in 3/15 (20.0 %), lymph node cytology in 1/15 (6.7 %), and blood PCR in 1/15 (6.7 %). All dogs treated had a complete resolution of diarrhea with specific treatment for leishmaniosis alone, based on meglumine antimoniate (75-100 mg/kg SID SC for 1 month) plus allopurinol (10 mg/kg BID PO ≥ 6 months). This study suggests that leishmaniosis should be also included in the differential diagnosis of dogs from endemic areas presenting with the primary problem of large-bowel, small-bowel, or mixed-bowel chronic diarrhea.

Prevalence and Characterization of Urinary Tract Infection in Owned Dogs and Cats From Spain.

Bacterial urinary tract infection (UTI) is a common clinical concern in dogs. However, incidence of feline UTI is much lower than in dogs although an increasing prevalence has been registered. The main objective of the present study was to describe and characterize the prevalence of urinary tract pathogens in urine samples of dogs and cats with urinary clinical signs throughout different Spanish provinces. Secondary aims were to determine if there were differences in urine sample characterizations based on species (i.e., dog and cat) or season. Dogs were found to have a higher rate of positive urinary cultures than cats (39.3% and 24.7% of the cultures submitted, respectively). The bacterial genera most commonly isolated in dogs were Escherichia spp. (45.3%), Proteus spp. (13.2%), Staphylococcus spp. (11%), and Enterococcus spp. (8.6%). Whereas in the feline population, Escherichia spp. (42.7%), Enterococcus spp. (22.2%), and Staphyloccoccus spp. (15.2%) were the most frequently isolated bacteria. The highest rates of positive urine cultures were registered in Melilla (70%), Zamora (66.7%), Teruel (64.3%), and Guadalajara (60%). Moreover, the proportion of positive urine cultures was not homogeneously distributed across provinces. Finally, some seasonality was found among most isolated bacterias. Enterococcus spp. was significantly more prevalent in summer, whereas Escherichia spp. and Proteus spp. were more commonly isolated in spring and Pseudomonas spp. in autumn.

PCR survey of vectorborne pathogens in dogs living in and around Barcelona, an area endemic for leishmaniasis.

Blood samples from 153 dogs living in and around Barcelona were assayed for Leishmania infantum and Ehrlichia, Anaplasma, Rickettsia, Bartonella, Hepatozoon, Babesia and Theileria species by PCR amplification of DNA, and the amplicons obtained were sequenced. The prevalence of the infectious agents was L infantum (29.4 per cent), Ehrlichia and Anaplasma species (4.0 per cent), Hepatozoon canis (3.3 per cent), Babesia canis vogeli (2.0 per cent), Babesia gibsoni (2.0 per cent), Babesia canis canis (1.3 per cent) and Theileria annae (0.7 per cent). Coinfections were present in seven of the dogs and they were significantly associated with L infantum infection (P=0.024). There was a significant correlation between clinical signs of illness and the load of L infantum.

Leptospira seroprevalence in owned dogs from Spain.

Leptospira is a widespread zoonosis that has been linked to transmission between dogs and humans. The main purposes were to determine the seroprevalence of anti-Leptospira serum antibody and to identify the most common serovars in dogs in Spain. This is a cross-sectional study with 1,310 records of canine Leptospira testing data from Spain since 2015 to 2017. Inclusion criteria were individual cases with MAT test results for 8 serovars (Bratislava, Icterohaemorrhagiae, Australis, Pomona, Grippotyphosa, Autumnalis, Canicola and Saxkoebing) and to have the zip code data. Three hundred and thirty-eight samples (25.8%; 95%CI 23.6-28.4) were seropositive (≥1:100). According to geographic areas, North had the highest seroprevalence (38.0%; 95%CI 28.9-47.1) followed by South (29.4%; 95%CI 20.1-38.8), Center (28.6%; 95%CI 24.3-33.0), Mediterranean (22.3%; 95%CI 19.1-25.6) and Northwest (22.2%; 95%CI 7.9-36.4). Seropositivity (MAT ≥1:100) was most common to serovars Icterohaemorrhagiae (19.4%; 95%CI 17.2-21.5) and Bratislava (8.5%; 95%CI 7.0-10.0), followed by Grippotyphosa (7.2%; 95%CI 5.8-8.6), Australis (6.4%; 95%CI 5.0-7.7), Autumnalis (5.0%; 95%CI 3.8-6.2), Pomona (4.5%; 95%CI 3.3-5.6), Canicola (3.4%; 95%CI 2.4-4.4) and Saxkoebing (0.8%; 95%CI 0.3-1.3). An association was found between positivity (MAT ≥1:100) and males (P = 0.003) and dogs that were 6 years old or older were at higher risk of exposure (P = 0.001; OR 4.61; 95%CI 1.86-11.43). This study has shown that dogs in Spain are commonly exposed to Leptospira infection and points out the necessity to control the prevalence of this severe widespread zoonosis in dogs and humans.

Endosymbionts carried by ticks feeding on dogs in Spain.

Studies on tick microbial communities historically focused on tick-borne pathogens. However, there is an increasing interest in capturing relationships among non-pathogenic endosymbionts and exploring their relevance for tick biology. The present study included a total of 1600 adult ticks collected from domestic dogs in 4 different biogeographical regions of Spain. Each pool formed by 1 to 10 halves of individuals representing one specific ticks species was examined by PCR for the presence of Coxiellaceae, Rickettsia spp., Rickettsiales, Wolbachia spp., and other bacterial DNA. Of the pools analyzed, 92% tested positive for endosymbiont-derived DNA. Coxiella spp. endosymbionts were the most prevalent microorganisms, being always present in Rhipicephalus sanguineus sensu lato (s.l.) pools. Rickettsia spp. DNA was detected in 60% of Dermacentor reticulatus pools and 40% of R. sanguineus s.l. pools, with a higher diversity of Rickettsia species in R. sanguineus s.l. pools. Our study reveals a negative relationship of Rickettsia massiliae with the presence of tick-borne pathogens in the same pool of ticks. An additional endosymbiont, 'Candidatus Rickettsiella isopodorum', was only detected in D. reticulatus pools. Data from this study indicate that dogs in Spain are exposed to several endosymbionts. Due to the importance of tick-borne pathogens, characterizing the role of endosymbionts for tick physiology and prevalence, may lead to novel control strategies.

Prevalence of Proteinuria in Owned Dogs from Italy: A Multicentric Study.

Even though proteinuria is related to different causes, when it is persistent and associated with inactive urinary sediment, it is primarily due to kidney disease. Early detection of proteinuria allows us to identify several pathological conditions. The aim of the study was screening a canine population not known as being proteinuric, by the urinary dipstick. The study was carried out in seven Italian veterinary clinics during a period of six weeks. Dogs were enrolled with no restriction of sex or age. Females in estrus, dogs with signs of genitourinary diseases, or those previously diagnosed with proteinuric nephropathy were excluded. Dogs were considered "nonproteinuric" (NP) in case of negative dipstick test or "suspected proteinuric" (SP), if positive at the dipstick. When possible, proteinuria was confirmed by UPC ratio. A total of 1156 dogs were evaluated: 414 were from northern Italy and 742 from southern Italy. Based on dipstick test, 655 (56.6%) dogs were NP, while 501 (43.3%) were SP. Among the NP dogs 225 out of 414 (54.3%) were in northern Italy and 430 of 742 (57.9%) in southern Italy. One hundred eighty-nine of 414 (45.7%) SP dogs were identified in northern Italy and 312 of 742 (42.1%) in southern Italy. No statistical difference was found between the North and the South of Italy. UPC was available in 412 out of 501 SP samples: proteinuria was confirmed in 263 (63.86%) samples. Results from our study showed a high percentage of suspected proteinuric dogs, apparently not affected by renal diseases, together with the absence of statistically significant differences based on geographical area.

Histopathological features of ocular leishmaniosis in the dog.

Canine leishmaniosis (CL) can present with multiple clinical signs and ocular disease is reported to occur in almost 25% of affected dogs. The purpose of the present study was to characterize the nature of inflammation within the eyes of dogs with leishmaniosis and to determine whether parasites were present in these lesions. Eyes from 60 dogs with confirmed leishmaniosis that died or were humanely destroyed over a 4 year period were included in the study. Sections of formalin-fixed globes were stained with haematoxylin and eosin (HE) and subjected to immunohistochemistry using a Leishmania-specific antibody. Clinically evident ocular signs were present in 15 of 60 dogs (13 bilaterally and 2 unilaterally). Thirty-five of 60 dogs received some form of anti-protozoal treatment. In 36 of 120 eyes (30%) a granulomatous inflammatory infiltrate was found and in 32 of 120 eyes (26.6%) the parasite was identified immunohistochemically within the globe. Ocular tissues affected, in order of frequency, were conjunctiva and limbus, ciliary body, iris, cornea, sclera and iridocorneal angle, choroid and the optic nerve sheath. Different microscopical patterns were defined in each of these structures. Leishmania organisms and associated inflammation can be found in different ocular tissues, accounting for some of the ocular clinical signs described for this disease.

Detection of Leishmania infantum by real-time PCR in a canine blood bank.

OBJECTIVES: Risk for transmission of Leishmania infantum from blood products has been largely demonstrated in human and veterinary literature. Appropriate screening of canine blood donors is important especially in an endemic area such as Barcelona (Spain). The purpose of this study was to evaluate the presence of L infantum DNA parasites by real-time quantitative PCR in our canine blood bank. METHODS: Samples from blood products obtained from 92 canine blood donors were assayed for L infantum by means of real-time PCR amplification and quantification. RESULTS: The prevalence of quantitative PCR-positive blood samples among healthy seronegative blood donors was 19.6 per cent. CLINICAL SIGNIFICANCE: The results of this study show that L infantum infection is common in canine blood donors and their blood products in an endemic area, despite a negative commercial serological screening for infectious diseases. Therefore, screening by PCR should be included in an integrated approach to evaluate L infantum infection among potential blood donors.

PCR evaluation of selected vector-borne pathogens in dogs with pericardial effusion.

OBJECTIVES: To investigate evidence for selected vector-borne pathogen infections in dogs with pericardial effusion living in a Mediterranean area in which several canine vector-borne diseases are endemic. MATERIALS AND METHODS: Archived EDTA blood (n=68) and pericardial fluid samples (n=58) from dogs with pericardial effusion (n=68) were included. Dogs without pericardial effusion examined for other reasons were included as controls (n=60). Pericardial effusion was classified as neoplastic in 40 dogs, idiopathic in 23 dogs and of unknown aetiology in 5 dogs. Real-time PCR was performed for Leishmania infantum, Ehrlichia/Anaplasma species, Hepatozoon canis, Babesia species, Rickettsia species and Bartonella species, and sequencing of PCR products from positive samples was used to confirm species specificity. RESULTS: Vector-borne pathogens were found in 18 dogs: 16 of 68 dogs with pericardial effusion (23·5%) and two of 60 control dogs (3·3%). Positive dogs demonstrated DNA of Leishmania infantum (n=7), Anaplasma platys (n=2, one dog coinfected with Leishmania infantum), Babesia canis (n=5), Babesia gibsoni (n=3) and Hepatozoon canis (n=2). Vector-borne pathogens were more commonly detected among dogs with pericardial effusion than controls (P=0·001). There was no relationship between aetiology of the pericardial effusion and evidence of vector-borne pathogens (P=0·932). CLINICAL SIGNIFICANCE: Vector-borne pathogens are often detected in dogs with pericardial effusion and require further investigation, especially in dogs with idiopathic pericardial effusion. PCR can provide additional information about the potential role of vector-borne pathogens in dogs with pericardial effusion living in endemic areas.

Polymerase chain reaction detection of Bartonella spp. in dogs from Spain with blood culture-negative infectious endocarditis.

OBJECTIVES: The presence of Bartonella spp. was detected by polymerase chain reaction (PCR) in dogs from Spain with blood culture-negative endocarditis. The aim of this study is to add information about canine infectious endocarditis in Europe. ANIMALS: Thirty dogs with naturally occurring blood culture-negative endocarditis were examined from 2010 to 2017 at three veterinary referral hospitals, located in northwest, northeast, and southeast of Spain. METHODS: It is a retrospective study. Medical records were reviewed to extract relevant data. Frozen or paraffin-embedded cardiac valve tissue and/or ethylenediamine tetraacetic acid blood samples were evaluated by PCR for the presence of Bartonella DNA. Positive results were sequenced to confirm the species. RESULTS: Polymerase chain reaction was positive for eight out of 30 dogs included (26.6%). Bartonella rochalimae, Bartonella vinsonii subsp. berkhoffii, and Bartonella koehlerae were detected in valve tissue or blood. CONCLUSIONS: Bartonella could be an important cause of blood culture-negative infectious endocarditis in dogs from Spain. The outcome for those dogs affected with Bartonella spp. was grave. Prompt empirical treatment with amoxicillin-clavulanate plus fluoroquinolones could be of value in cases of blood culture-negative endocarditis.

Successful medical management of pseudomembranous cystitis in three cats with lower urinary tract obstruction.

CASE REPORT: The present case series describes the clinical course and outcome of three cats diagnosed with pseudomembranous cystitis. This is an uncommon presentation of lower urinary tract obstruction but can be easily be identified by ultrasonography, revealing severe bladder wall thickening and thin hyperechoic luminal strips. The condition can be secondary to severe bacterial urinary tract infection. All cats were successfully treated with medical management only, mainly based on antimicrobials and individualised supportive therapy. CONCLUSION: Further evaluation of this condition is necessary in order to determine potential underlying aetiologies, pathophysiological mechanisms and the most appropriate standardised treatment.

Advantages of real-time PCR assay for diagnosis and monitoring of canine leishmaniosis.

The aim of the present study is to highlight the advantages of real-time quantitative PCR intended to aid in the diagnosis and monitoring of canine leishmaniosis. Diagnosis of canine leishmaniosis is extremely challenging, especially in endemic areas, due to the diverse and non-specific clinical manifestations, and due to the high seroprevalence rate in sub-clinical dogs. Veterinarian clinicians are usually confronted with cases that are compatible with the disease, and with several diagnostic tests, sometimes with contradictory results. We have developed a new TaqMan assay, targeting the kinetoplast, applied to 44 samples of bone marrow aspirate or peripheral blood. The dynamic range of detection of Leishmania DNA was established in 7 logs and the limit of detection is 0.001 parasites in the PCR reaction. At the time of diagnosis parasitemia ranges from less than 1 to 10(7)parasites/ml. The ability to quantify the parasite burden allowed: (i) to elucidate the status of positive dogs by conventional PCR, although larger studies are necessary to clarify the dividing line between infection and disease, (ii) to estimate the kinetics of the parasite load and the different response to the treatment in a follow-up and (iii) to validate blood as less invasive sample for qPCR. The continuous data provided by real-time qPCR could solve the dilemma for the clinician managing cases of canine leishmaniosis by differentiating between Leishmania-infected dogs or dogs with active disease of leishmaniosis.

The effect of renal diet in association with enalapril or benazepril on proteinuria in dogs with proteinuric chronic kidney disease.

Treating proteinuria in dogs reduces the progression of chronic kidney disease (CKD); renal diets and angiotensin-converting enzyme (ACE)-inhibitors are cornerstones of treatment. Whether different ACE-inhibitors have distinct kidney protective effects is unknown; it is therefore hypothesized that renal diets and enalapril or benazepril have different beneficial effects in proteinuric CKD dogs. Forty-four dogs with proteinuric CKD (IRIS stages 1-4) were enrolled in the study and were fed renal diet for 30 days. Thereafter, they were randomly assigned to one of 2 groups. Dogs in group A (n=22) received enalapril (0.5 mg/kg, q12h) and in group B (n=22) benazepril (0.5 mg/kg, q24h); in both groups, dogs were fed the same renal diet. After randomization, dogs were monitored for 120 days. Body weight and body condition score (BCS), serum concentrations of creatinine, blood urea nitrogen (BUN), albumin and total proteins, and urine protein-to-creatinine (UPC) ratio were compared at different time-points. After 30 days of renal diet, creatinine, BUN and UPC ratio decreased significantly (p<0.0001). Compared to randomization, body weight, BCS, albumin, total proteins, creatinine and BUN did not vary during follow-up in the 44 dogs and differences between group A and B were not observed. However, the UPC ratio of group A at day 60, 90 and 150 was significantly lower than in group B and compared to randomization (p<0.05). In group B it did not vary overtime. It is concluded that the renal diet is beneficial to decrease creatinine, BUN and UPC ratio in proteinuric CKD dogs. Enalapril further ameliorates proteinuria if administered along with renal diet.

Adverse urinary effects of allopurinol in dogs with leishmaniasis.

OBJECTIVE: The objective of this study was to describe the adverse effects of allopurinol on the urinary system during treatment of canine leishmaniasis. METHODS: Retrospective case series of 42 dogs that developed xanthinuria while receiving allopurinol treatment for leishmaniasis. RESULTS: Of 320 dogs diagnosed with leishmaniasis, 42 (13%) developed adverse urinary effects. Thirteen (of 42) dogs (31%) developed xanthinuria, renal mineralisation and urolithiasis; 11 (26·2%) showed xanthinuria with renal mineralisation; 9 (21·4%) had xanthinuria with urolithiasis and 9 (21·4%) developed xanthinuria alone. Urinary clinical signs developed in 19 dogs (45·2%). CLINICAL SIGNIFICANCE: This study demonstrates that urolithiasis and renal mineralisation can occur in dogs receiving allopurinol therapy. Dogs receiving therapy should be monitored for the development of urinary adverse effects from the beginning of treatment.

Clinical leishmaniasis in dogs living in the UK.

OBJECTIVES: To investigate the prevalence of leishmaniasis in dogs in the UK and to describe clinical presentation, clinicopathological abnormalities, therapeutic protocols and outcome in this non-endemic country. MATERIALS AND METHODS: Medical records of dogs diagnosed with leishmaniasis at seven referral centres in the UK were retrospectively reviewed. RESULTS: The prevalence was between 0·007 and 0·04% with a higher number of cases in southern England. All dogs had a history of travel to or from an endemic country. Lethargy, dermatological disease, decreased appetite and lameness were the most common reasons for presentation. Allopurinol was used alone for treatment in the majority of cases. CLINICAL SIGNIFICANCE: Although rare, leishmaniasis should be considered in dogs in the UK if they have compatible clinical signs and history of travel to or from endemic areas.

Diagnostic potential of Western blot analysis of sera from dogs with leishmaniasis in endemic areas and significance of the pattern.

Serum samples collected from 237 dogs in Catalonia (northeastern Spain) were screened by Western blot analysis to detect the presence of antibodies specific to different Leishmania infantum polypeptide fractions. Leishmaniasis was confirmed in 72 of these dogs by direct examination and/or culture. Another 165 animals from the Priorat region were studied periodically for 2-8 years between 1987 and 1995, giving a total of 565 determinations. A control group of 93 dogs from nonendemic areas was also studied. Sera from dogs with leishmaniasis recognized antigens with molecular weights ranging from 12 to 85 kD. The most sensitive antigens were those of 70, 65, 46, 30, 28, 14, and 12 kD, which were recognized by 75%, 75%, 78%, 75%, 81%, 79%, and 75%, respectively, of the sera from dogs with positive parasitologic examination results. Antigens of 70 and 65 kD were also recognized by two dogs from nonendemic areas. Antigens of 14 and 12 kD were the first to be recognized by sera of asymptomatic dogs with titers less than the cut-off value of the dot-ELISA that increased during the longitudinal study, and the presence of antibodies specific for these fractions was observed for up to six years before seroconversion observed by dot-ELISA. These antibodies were also the first to disappear in dogs in which the disease was self-limited. The study corroborates the high sensitivity and specificity of Western blots in the diagnosis of canine leishmaniasis when the bands of low molecular weight (less than 46 kD) are considered, and indicates that fractions of 14 and 12 kD are useful in detecting early forms of the disease.

Leishmania infantum-specific IgG, IgG1 and IgG2 antibody responses in healthy and ill dogs from endemic areas. Evolution in the course of infection and after treatment.

The expression of IgG, IgG1 and IgG2 specific antibodies for Leishmania infantum was studied in five groups of dogs in Catalonia (Spain): I, 99 asymptomatic dogs (infected and uninfected) from a highly endemic area for leishmaniosis; II, 139 untreated dogs with clinically patent leishmaniosis; III, 11 naturally infected asymptomatic dogs monitored for up to 5 years since they were found seropositive to Leishmania antigen and without treatment; IV, 25 naturally infected dogs with clinically patent leishmaniosis and treated with either meglumine antimoniate and allopurinol or allopurinol alone and V, six experimentally infected dogs, treated with meglumine antimoniate and controlled for 5 years. The levels (ELISA units) of IgG, IgG1 and IgG2 in asymptomatic dogs (group I) were very variable (24+/-33, 32+/-31 and 26+/-31, respectively), and, as expected, lower than in ill dogs (group II) (168+/-34, 84+/-71 and 172+/-31, respectively). In both groups, the correlation between IgG and IgG2 levels (r=0.95, P<0.001 in group I and r=0.63, P<0.001 in group II) was higher than between IgG and IgG1 levels (r=0.01, P>0.05 in group I and r=0.31, P<0.001 in group II). In group III, IgG and IgG2 expression increased during infection, while IgG1 expression remained the same. In dogs of group IV, IgG levels after 1 year of treatment decreased more in responsive (mean values, 163+/-42 before treatment (b.t.) and 100+/-36 after treatment (a.t.)) than in unresponsive dogs (158+/-29 b.t. and 124+/-51 a.t.), especially for IgG1 (94+/-89 b.t. and 20+/-21 a.t. in responsive dogs and 35+/-25 b.t. and 22+/-13 a.t. in unresponsive dogs) rather than for IgG2 (156+/-16 b.t. and 114+/-45 a.t. in responsive and 151+/-11 b.t. and 125+/-36 a.t. in unresponsive dogs). Similar results were observed in the evolution of experimentally infected animals after consecutive and specific treatments. Overall results show the great variation in Leishmania-specific IgG1 expression in asymptomatic and symptomatic dogs, their lack of correlation with that of IgG2 and chemotherapy is more effective in dogs with initially high expression of IgG1.