Vascular permeability and coagulation during Rickettsia rickettsii infection in dogs

The vascular permeability of the ocular fundus, alterations in the coagulation system, and plasma concentrations of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) were studied in dogs following intradermal inoculation with 5 x 10(5) TCID50 of Rickettsia rickettsii. Twenty-four to 48 hours after the onset of fever and rickettsemia, multifocal areas of retinal vasculitis were evident, which corresponded to areas of altered vascular permeability demonstrated by fluorescein angiography. The number and intensity of retinal vessels with sodium fluorescein leakage peaked during the second week after inoculation, and retinal vascular permeability remained altered during the third week of infection, well past the phase of clinical and clinicopathologic recovery. Development of retinal vasculitic foci was associated with thrombocytopenia, increased concentrations of circulating fibrinogen, and slight prolongation of activated partial thromboplastin time. Increased concentrations of fibrin/fibrinogen degradation products were detected in 4 of 9 dogs. Despite the degree of vascular endothelial damage evident on fluorescein angiographic and histologic studies in these dogs, plasma TXB2 and 6-keto-PGF1 alpha concentrations were not increased.     

Platelet aggregation in dogs experimentally infected with Rickettsia rickettsii

Platelet aggregation studies were performed on nine Beagle dogs experimentally infected with Rickettsia rickettsia. Platelets from dogs with Rocky Mountain spotted fever tended to be more aggregable than controls.     

An unmatched case controlled study of clinicopathologic abnormalities in dogs with Bartonella infection

We compared clinicopathologic findings in dogs with Bartonella infection to Bartonella spp. negative dogs suspected of a vector-borne disease. Cases (n = 47) and controls (n = 93) were selected on the basis of positive or negative enrichment culture PCR results, respectively. Signalment, clinicopathologic findings and treatments were extracted from medical records. DNA sequencing identified Bartonella henselae (n = 28, 59.6%), Bartonella vinsonii subsp. berkhoffii (n = 20, 42.6%), Bartonella koehlerae (n = 3, 6.4%), Bartonella volans-like (n = 3, 6.4%) and Bartonella bovis (n = 1, 2.1%). There were no significant differences in age, breed, size, sex or neuter status between cases and controls. Dogs infected with Bartonella sp. often had a history of weight loss [OR = 2.82; 95% CI: 1.08–7.56] and were hypoglobulinemic [OR = 4.26; 95% CI: 1.31–14.41]. With the exception of weight loss and hypoglobulinemia, clinicopathologic abnormalities in Bartonella-infected dogs in this study were similar to dogs suspected of other vector-borne infections.     

Kinetics of IgM and IgG responses to experimental and naturally acquired Rickettsia rickettsii infection in dogs

The kinetics of specific IgM and IgG antibody response was characterized in four 9-month-old Beagles after inoculation of 2 x 10(2) plaque-forming units (PFU) of Sheila Smith strain of Rickettsia rickettsii. Immunoglobulin M antibodies were first detected by indirect immunofluorescence on postinoculation (PI) day 9, peaked by PI day 20, and were no longer detectable by PI day 80. Immunoglobulin G antibodies became detectable between PI days 22 and 28, peaked by PI day 42, and decreased gradually through PI day 130. Subsequent challenges with R rickettsii on PI days 216 (2 x 10(2) PFU/dog) and 1,029 (5 x 10(4) tissue culture infective dose [TCID50]/dog) resulted in slightly different serologic responses. The initial challenge exposure failed to increase the concentration of IgG antibodies and induced only low concentrations of IgM antibodies. After the second challenge inoculation, IgM antibodies were not detectable and the concentration IgG antibodies increased slightly. Clinical abnormalities and seroconversion were documented in control dogs following each challenge exposure. Examination of acute and convalescent serum samples from 55 dogs in which Rocky Mountain spotted fever was suspected clinically suggested that sole evaluation of IgM antibodies in acute-phase serum would result in inaccurate diagnoses because of false-positive and -negative results. Use of a composite conjugate that detects IgM and IgG antibodies to R rickettsii appears to be satisfactory for diagnostic purposes; however, concurrent quantitation of IgM antibodies may facilitate serodiagnosis in a select group of dogs in which a four-fold increase in convalescent antibody titer is not detected by use of the composite conjugate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Clinical, hematologic, and humoral immune response in female dogs inoculated with Rickettsia rickettsii and Rickettsia montana

Female Beagles were inoculated intradermally with a sublethal dose of Rickettsia rickettsii and R montana. Three dogs (group 1) were inoculated with 2 X 10(2) plaque-forming units (PFU) of R rickettsia and were treated with tetracycline beginning on postinoculation day (PID) 12; 3 dogs (group 2) were inoculated with 2 X 10(2) PFU of R rickettsii but were not treated; 3 dogs (group 3) were inoculated with 2 X 10(2) PFU of R montana. Group-3 dogs failed to seroconvert and were inoculated a second time on PID 68. Groups 1 and 2 dogs inoculated with R rickettsii became depressed and developed occasional inappetence, fever, hematochezia, and ocular lesions. These dogs had a decrease in PCV and RBC count, an initial decrease in WBC count followed by leukocytosis, and a decrease in platelet count. Group-3 dogs inoculated with R montana remained healthy. After R rickettsii inoculation, the serologic response to spotted fever group (SFG) rickettsial antigens (R rickettsii, R rhipicephali, R montana, and R bellii) was similar. The antibody response to R rickettsii was first detected on PID 9, with peak titers reached by PID 20. Serum titers to R rickettsii remained stable or decreased one dilution through PID 120. Of 4 SFG rickettsial antigens, the highest serologic response was to R rickettsii. A cross-reacting antibody response with R rhipicephali and R montana was nearly identical and was only slightly less than the response to R rickettsii. Cross-reacting antibodies to R belli were of lower mean titer and of shorter duration than were cross-reacting antibodies to other SFG rickettsiae.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular survey of Rickettsia spp. in sick dogs in Italy

The aim of this retrospective study was to investigate the prevalence of Rickettsia spp. DNA in the blood of sick dogs from Italy. Canine blood samples (n=650) submitted for molecular testing of Rickettsia spp. to a diagnostic laboratory from February 2003 to March 2006 were studied. The Rickettsia spp. DNA detection was performed by Light Cycler real-time PCR using hybridization probes separately conducted with specific primers and probes. The total percentage of Rickettsia spp.-positive dog samples was 1.5% (10 out of 650). The percentage of Rickettsia spp.-positive dog samples submitted from north, central and southern Italy were 0.4% (1/248), 1.4% (3/219) and 3.3% (6/183), respectively. Five out of 138 dogs (3.6%) from Sicily were positive on Rickettsia PCR testing. A statistical difference was found between the percentages of positive samples from the Yorkshire terrier group (10.7%) compared with the mixed breed group (0.7%). No statistical differences were found between seasonal period, region and gender. Based on molecular data, there is infrequent rickettsiemia in dogs.     

Investigation of Bartonella henselae in cats in Ankara, Turkey

The purpose of this study was to determine Bartonella henselae prevalance in cats in Ankara. Whole bloods and sera collected from 256 cats were investigated for the presence feline Bartonella species by culture and sera were tested for the presence of antibodies against B. henselae IgG using immunofluorescence assay. Bartonella species were isolated by blood culture from 24 (9.4%) cats. Bartonella isolates were subjected to restriction fragment length polymorphism (RFLP) by using TaqI and HhaI endonucleases to identify species. Twenty-one isolates were determined as B. henselae and three of 24 isolates were determined as Bartonella clarridgeiae with RFLP. The bacteraemia prevalence and seroprevalence of B. henselae IgG antibodies in cats was detected as 8.2% and 18.6% respectively. This is the first report on B. henselea and B. clarridgeiae in cats in Turkey.     

Granulomatous disease associated with Bartonella infection in 2 dogs

Shortly after removal of an engorged tick from the left ear, a 4-year-old Greyhound was referred for evaluation of fever and a rapidly enlarging mass in the region of the left submandibular lymph node. Histopathologic evaluation of the lymph node resulted in a diagnosis of severe granulomatous lymphadenitis. An 11-year-old mixed-breed dog was referred for evaluation of a 6-week history of serous nasal discharge. Histologic examination of a surgical biopsy from a nasal mass indicated multifocal granulomatous inflammation with fibrosis. Serum samples obtained from both dogs were reactive by immunofluorescent assay to Bartonella vinsonii subsp. berkhoffii antigens (reciprocal titers of 128). Although Bartonella organisms were not isolated by lysis centrifugation blood culture, Bartonella DNA was amplified from tissue samples obtained from each dog (lymph node biopsy from dog 1 and nasal biopsy from dog 2) using primers that amplify a portion of the 16S rRNA gene followed by Southern blot hybridization using a genus-specific probe. Additionally, restriction fragment length polymorphism (RFLP) analysis of a Bartonella-specific citrate synthase gene product obtained from dog 2 resulted in a restriction pattern identical to B. vinsonii subsp. berkhoffii. This is the 1st report of granulomatous disease in dogs associated with Bartonella infection.     

Detection of Bartonella spp. in Cimex lectularius by MALDI-TOF MS

Bed bugs are small hematophagous insects. They are found in temperate and tropical climates around the world. Their vectorial capacity for several pathogens, including Bartonella spp., has been suspected. An experimental study of artificial infection of Cimex lectularius with Bartonella quintana and Bartonella henselae bacteria was developed to evaluate the ability of MALDI-TOF MS to simultaneously identify bed bugs and their infectious status. This experimental study confirmed the ability of MALDI-TOF MS to identify bed bugs. In addition, it was able to differentiate between control bed bugs, bed bugs infected with Bartonella quintana and bed bugs infected with Bartonella henselae, with an identification percentage above 90%.     

Flea species infesting dogs in Florida and Bartonella spp. prevalence rates

Several Bartonella spp. associated with fleas can induce a variety of clinical syndromes in both dogs and humans. However, few studies have investigated the prevalence of Bartonella in the blood of dogs and their fleas. The objectives of this study were to determine the genera of fleas infesting shelter dogs in Florida, the prevalence of Bartonella spp. within the fleas, and the prevalence of Bartonella spp. within the blood of healthy dogs from which the fleas were collected. Fleas, serum, and EDTA-anti-coagulated whole blood were collected from 80 healthy dogs, and total DNA was extracted for PCR amplification of Bartonella spp. The genera of fleas infesting 43 of the dogs were determined phenotypically. PCR amplicons from blood and flea pools were sequenced to confirm the Bartonella species. Amplicons for which sequencing revealed homology to Bartonella vinsonii subsp. berkhoffii (Bvb) underwent specific genotyping by targeting the 16S–23S intergenic spacer region.A total of 220 fleas were collected from 80 dogs and pooled by genus (43 dogs) and flea species. Bartonella spp. DNA was amplified from 14 of 80 dog blood samples (17.5%) and from 9 of 80 pooled fleas (11.3%). B. vinsonii subsp. berkhoffii DNA was amplified from nine dogs and five of the flea pools. Bartonella rochalimae (Br) DNA was amplified from six dogs and two flea pools. One of 14 dogs was co-infected with Bvb and Br. The dog was infested with Pulex spp. fleas containing Br DNA and a single Ctenocephalides felis flea. Of the Bvb bacteremic dogs, five and four were infected with genotypes II and I, respectively. Of the Bvb PCR positive flea pools, three were Bvb genotype II and two were Bvb genotype I.Amplification of Bvb DNA from Pulex spp. collected from domestic dogs, suggests that Pulex fleas may be a vector for dogs and a source for zoonotic transfer of this pathogen from dogs to people. The findings of this study provide evidence to support the hypothesis that flea-infested dogs may be a reservoir host for Bvb and Br and that ectoparasite control is an important component of shelter intake protocols.     

Bartonella spp. DNA in cardiac tissues from dogs in Colorado and Wyoming

Background: Several Bartonella species (spp.) have been identified in dogs diagnosed with infectious endocarditis (IE) or myocarditis. Objective: To interrogate cardiac tissues of dogs with suspected IE for the presence of Bartonella spp. DNA of dogs in the Rocky Mountain states. Animals: Nine dogs with a clinical diagnosis of endocarditis from January 1990 to June 2008 were included. Methods: In this retrospective study, medical records at the Veterinary Teaching Hospital were searched. Animals were excluded if there was no diagnosis of IE in the original necropsy report. Paraffin embedded tissue blocks and medical records were available from 9 dogs. Total DNA was extracted from the cardiac tissues and assessed for Bartonella spp. DNA by 3 polymerase chain reaction (PCR) methods. For positive samples, the Bartonella spp. were determined by genetic sequencing or fluorogenic real‐time PCR. Results: Bartonella henselae DNA was amplified from the tissues of 7 dogs; Bartonella vinsonii subsp berkhoffii DNA was amplified concurrently from 3 dogs. Six dogs were from Colorado and 1 was from Wyoming. Flea or tick infestations were reported in 2 dogs. Conclusions and Clinical Importance: Bartonella spp. should be on the differential list for dogs in the Rocky Mountain states. The results emphasize the need for routine use of external parasite control products even in regions perceived to have low risk for flea and tick infestations.     

Detection of Bartonella henselae DNA in two dogs with pyogranulomatous lymphadenitis

CASE DESCRIPTION: 1 dog evaluated because of inappetence and lameness of the left hind limb of 1 day's duration and 1 dog evaluated because of inappetence, fever, and lymphadenopathy of 2 weeks' duration. CLINICAL FINDINGS: Histologic examination of excisional biopsy specimens from lymph nodes revealed pyogranulomatous lymphadenitis in both dogs. Quantitative real-time PCR assays detected Bartonella henselae DNA in blood samples and affected lymph node specimens from both dogs. Antibodies against B. henselae were not detected via immunofluorescent antibody testing during active disease in either dog. TREATMENT AND OUTCOME: 1 dog recovered after 6 weeks of treatment with doxycycline (5 mg/kg [2.3 mg/lb], p.o., q 12 h), whereas the other dog recovered after receiving a combination of azithromycin (14.5 mg/kg [6.6 mg/lb], p.o., q 24 h for 21 days), doxycycline (17.3 mg/kg [7.9 mg/lb], p.o., q 24 h for 4 weeks), and immunosuppressive corticosteroid (prednisone [3 mg/kg {1.4 mg/lb}, p.o., q 24 h], tapered by decreasing the daily dose by 25% every 2 weeks) treatment. CLINICAL RELEVANCE: B. henselae is implicated as a possible cause or a cofactor in the development of pyogranulomatous lymphadenitis in dogs. In dogs with pyogranulomatous lymphadenitis, immunofluorescent assays may not detect antibodies against B. henselae. Molecular testing, including PCR assay of affected tissues, may provide an alternative diagnostic method for detection of B. henselae DNA in pyogranulomatous lymph nodes.     

Bartonella bovis and Bartonella chomelii infection in dairy cattle and their ectoparasites in Algeria

Bartonella are blood-borne and vector-transmitted bacteria, some of which are zoonotic. B. bovis and B. chomelii have been reported in cattle. However, no information has yet been provided on Bartonella infection in cattle in Algeria. Therefore, 313 cattle from 45 dairy farms were surveyed in Kabylia, Algeria, in order to identify Bartonella species infecting cattle using serological and molecular tests. In addition, 277 ticks and 33 Hippoboscidae flies were collected. Bartonella bovis and B. chomelii were identified as the two species infecting cattle. Bartonella DNA was also amplified from 6.8 % (n = 19) of ticks and 78.8 % (n = 26) of flies. Prevalence of B. bovis DNA in dairy cattle was associated both with age and altitude. This study is the first one to report of bovine bartonellosis in Algeria, both in dairy cattle and in potential Bartonella vectors, with the detection of B. bovis DNA in tick samples and B. chomelii in fly samples.     

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.

Detection of Bartonella henselae and Bartonella clarridgeiae DNA in hepatic specimens from two dogs with hepatic disease

A 4-year-old Basset Hound and a 6-year-old Doberman Pinscher were referred for diagnostic evaluation following documentation of persistently increased hepatic enzyme activities and hepatic dysfunction. Histologic evaluation of hepatic biopsy specimens from the 2 dogs revealed granulomatous hepatitis in the Basset Hound and lymphocytic hepatitis with fibrosis and copper accumulation in the Doberman Pinscher. No etiologic agents were identified histologically. Bartonella henselae DNA was subsequently amplified from hepatic tissue from the Basset Hound and Bartonella clarridgeiae was amplified from hepatic tissue from the Doberman Pinscher. Amplification was performed with a polymerase chain reaction assay incorporating primers that target a portion of the 16S-23S rRNA intergenic spacer region. Both dogs were treated with azithromycin, in combination with a variety of other medications and herbal treatments, and improved clinically. Identification of Bartonella DNA in these dogs indicates the need for future prospective studies to determine the clinical relevance of Bartonella spp infection in dogs with hepatic disease.     

Prevalence of Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, Bartonella vinsonii berkhoffii, and Rickettsia spp. in dogs from Grenada

To identify the tick-borne pathogens in dogs from Grenada, we conducted a serologic survey for Ehrlichia canis in 2004 (104 dogs) and a comprehensive serologic and molecular survey for a variety of tick-borne pathogens in 2006 (73 dogs). In 2004 and 2006, 44 and 32 dogs (42.3% and 43.8%) were seropositive for E. canis, respectively. In 2006, several tick-borne pathogens were identified by serology and PCR. DNA of E. canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, and Bartonella sp. were identified in 18 (24.7%), 14 (19.2%), 5 (7%), 5 (7%), and 1 (1.4%) dogs, respectively. Six (8.2%) dogs were seropositive for Bartonella vinsonii subsp. berkhoffii. All dogs were seronegative and PCR-negative for Rickettsia spp. Coinfection with two or three pathogens was observed in eight dogs. Partial 16S rRNA E. canis and A. platys sequences were identical to sequences in GenBank. Partial 18S rRNA gene sequences from the Grenadian H. canis were identical to each other and had one possible mismatch (ambiguous base) from H. canis detected from Spain and Brazil. Grenadian B. c. vogeli sequences were identical to B. c. vogeli from Brazil and Japan. All of the detected pathogens are transmitted, or suspected to be transmitted, by Rhipicephalus sanguineus. Results of this study indicate that dogs from Grenada are infected with multiple tick-borne pathogens; therefore, tick-borne diseases should be included as differentials for dogs exhibiting thrombocytopenia, leukopenia, fever, or lethargy. One pathogen, E. canis, is also of potential public health significance.     

Gastric function in dogs with naturally acquired gastric Helicobacter spp. infection

The association of Helicobacter pylori with gastritis, peptic ulcers, and gastric neoplasia has led to fundamental changes in the understanding of gastric disease in humans. The relationship of Helicobacter spp. infection to gastric disease in dogs is unclear. The objective of this study was to determine if Helicobacter infection affects the gastric secretory axis of dogs. Eight Beagle dogs with naturally acquired Helicobacter spp. infection were studied before and after (4 and 29 days) the attempted eradication of Helicobacter spp. with a combination of amoxicillin, metronidazole, and famotidine (AMF). Six specific-pathogen-free, Helicobacter-free Beagle dogs served as controls. The electron microscopic appearance of spiral organisms in infected dogs indicated coinfection with Helicobacter felis- and H bizzozeronii-like organisms. Unstimulated gastric pH and fasting, postprandial, and bombesin-stimulated plasma gastrin were similar in both infected and uninfected dogs, although a trend (P = .09) toward higher meal-stimulated gastrin was observed in infected dogs at 60 minutes. Pentagastrin-stimulated maximal acid output (mmol HCI/kg0.75/hour) and titratable acidity (mmol HCl/mL) were similar in both infected and uninfected dogs, but gastric pH during maximal acid output was lower (P < .01) in uninfected dogs. Mild gastric inflammation was present in both infected and uninfected dogs. Gastric spiral organisms were undetectable in 6/8 infected dogs 4 days after AMF but had recurred in 8/8 dogs 29 days after AMF. Analysis of gastric DNA with Helicobacter-specific primers indicated persistence of Helicobacter DNA at 4 and 29 days after antibiotic therapy. Acid secretion, plasma gastrin, and mucosal inflammation were not affected by the transient suppression of Helicobacter spp. by AMF. These findings suggest that gastric secretory function in dogs is not markedly perturbed by naturally acquired Helicobacter spp. infection and that treatment with amoxicillin, metronidazole, and famotidine causes suppression rather than eradication of gastric Helicobacter spp. in dogs.     

Detection of platelet-bound antibodies in beagle dogs after artificial infection with Ehrlichia canis

Six dogs were infected with Ehrlichia canis by intravenous injection of heavily infected DH82 cells. All dogs developed typical signs of canine monocytic ehrlichiosis. Using flow cytometric technology, platelet-bound IgG (PBIgG) were detected in 5 of the 6 dogs after experimental infection with E. canis over a period of 3-10 days post infection (PI). The first detection of PBIgG was made as early as day 3 PI in 2 out of 6 dogs, and on day 5 PI in 1 dog. On day 7 PI, PBIgG was detected in 2 dogs, and on day 10 PI in 3 out of 6 dogs. This is the first report documenting the presence of PBIgG following E. canis infection in dogs. This finding further supports the theory that the thrombocytopenia seen in canine monocytic ehrlichiosis has an immunological component and that exposure to an infectious agent, in this case the rickettsia E. canis, can trigger autoimmune mechanisms. Due to the heterogeneous appearance of PBIgG among the infected dogs it was concluded that other non-immunological mechanisms are probably also involved in the pathogenesis of the thrombocytopenia seen in canine monocytic ehrlichiosis.     

Bartonella spp infection as a possible cause of uveitis in a cat

A 6-year-old castrated mixed-breed cat was evaluated because of unilateral anterior uveitis. The cat was seronegative for antibodies to Toxoplasma gondii, coronaviruses, and feline immunodeficiency virus, and antigens for FeLV p27 and Cryptococcus neoformans. Antibodies to Bartonella spp were detected in serum and aqueous humor. The antibody coefficient (C value) for IgG antibodies to Bartonella spp in the aqueous humor was 4.42; values > 1 suggest ocular production of antibodies and supports a diagnosis of ocular infection. Topical administration of prednisolone and oral administration of prednisone failed to induce a response; however, the uveitis resolved rapidly after the cat was given doxycycline orally. Clinical or laboratory evidence of immunodeficiency in this cat was not detected. Detection of a serum IgG antibody titer to Bartonella spp and ocular production of IgG antibodies to Bartonella spp, exclusion of other causes of uveitis, and response to doxycycline suggests that the cat may have had bartonellosis resulting in uveal tract inflammation.