Approaches Used to Construct Antibiograms for Dogs in a Veterinary Teaching Hospital in the United States.

Non-judicious antimicrobial use (AMU) is a major driver of antimicrobial resistance (AMR). In human hospitals, cumulative antibiograms are often used by clinicians to evaluate local susceptibility rates and to select the most appropriate empiric therapy with the aim of minimizing inappropriate AMU. However, the use of cumulative antibiograms to guide empiric antimicrobial therapy in veterinary hospitals in the United States is limited, and there are no specific guidelines or standardized methods available for the construction of antibiograms in veterinary clinical settings. The objective of this methods article is to describe the approaches that were used to construct antibiograms from clinical samples collected from dogs seen at a veterinary teaching hospital. Laboratory data for 563 dogs for the period from 1 January 2015 to 31 December 2015 was utilized. We used the Clinical and Laboratory Standards Institute (CLSI) guidelines for use in the construction of the antibiograms in human healthcare settings as the basis for the veterinary antibiograms. One general antibiogram and antibiograms stratified by hospital section, the anatomic region of sample collection/by sample type, were created and the challenges encountered in preparing these antibiograms were highlighted. The approaches described could be useful in guiding veterinary antibiogram development for empiric therapy.

Antimicrobial Susceptibility of Bacteria Isolated from Freshwater Mussels in the Wildcat Creek Watershed, Indiana, United States.

Antimicrobial resistance (AMR) is a global health crisis that threatens the health of humans and animals. The spread of resistance among species may occur through our shared environment. Prevention of AMR requires integrated monitoring systems, and these systems must account for the presence of AMR in the environment in order to be effective. The purpose of this study was to establish and pilot a set of procedures for utilizing freshwater mussels as a means of surveillance for microbes with AMR in Indiana waterways. One hundred and eighty freshwater mussels were sampled from three sites along the Wildcat Creek watershed in north-central Indiana. Specimens were evaluated for the presence of ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species), Escherichia coli, Campylobacter, and Salmonella species, and the isolates were tested for antimicrobial resistance. A total of 24 bacterial isolates were obtained from tissue homogenates of freshwater mussels collected at a site directly downstream from Kokomo, Indiana. Of these, 17 were Enterobacter spp., five were Escherichia coli, one was Pseudomonas aeruginosa, and one was Klebsiella pneumoniae. All isolates were resistant to three or more antimicrobial drug classes. Further work is necessary to determine the source of the bacterial species found in the mussels.

Nested PCR Detection of Pythium sp. from Formalin-Fixed, Paraffin-Embedded Canine Tissue Sections.

Pythium insidiosum is an infectious oomycete affecting dogs that develop the cutaneous or gastrointestinal form of pythiosis with a poor prognosis. If left untreated, pythiosis may be fatal. This organism is not a true fungus because its cell wall and cell membrane lack chitin and ergosterol, respectively, requiring specific treatment. Identifying the organism is challenging, as a hematoxylin and eosin (H&E) stain poorly stain the P. insidiosum hyphae and cannot be differentiated conclusively from other fungal or fungal-like organisms (such as Lagenidium sp.) morphologically. Our study aimed to develop a nested PCR to detect P. insidiosum and compare it with the traditional histopathologic detection of hyphae. Formalin-fixed, paraffin-embedded (FFPE) tissue scrolls from 26 dogs with lesions suggesting the P. insidiosum infection were assessed histologically, and DNA was extracted from the FFPE tissue sections for nested PCR. Agreement between the histologic stains, (H&E), periodic acid-Schiff (PAS), and/or Grocott methenamine silver (GMS) and the nested PCR occurred in 18/26 cases. Hyphae consistent with Pythium sp. were identified via histopathology in 57.7% of the samples, whereas the nested PCR detected P. insidiosum in 76.9% of samples, aiding in the sensitivity of the diagnosis of pythiosis in dogs. Using this combination of techniques, we report 20 canine cases of pythiosis over 18 years in Indiana and Kentucky, an unexpectedly high incidence for temperate climatic regions. Using a combination of histopathology evaluation and nested PCR is recommended to aid in the accurate diagnosis of pythiosis.

Antimicrobial susceptibility and risk factors for resistance among Escherichia coli isolated from canine specimens submitted to a diagnostic laboratory in Indiana, 2010-2019.

Escherichia coli (E. coli) is the most common Gram-negative pathogen isolated in human infections. Antimicrobial resistant (AMR) E. coli originating from dogs may directly or indirectly cause disease in humans. The objective of this study was to calculate the proportion of antimicrobial susceptible E. coli isolated from canine specimens submitted to the Indiana Animal Disease Diagnostic Laboratory and to identify temporal patterns of susceptibility among these isolates. Susceptibility data of 2,738 E. coli isolates from dogs from 2010 through 2019 were used in this study. Proportions of isolates susceptible to the various antimicrobials were calculated using SAS statistical software and the Cochran-Armitage trend test was used to investigate the temporal trends in susceptibility. A multivariable binary logistic regression model was built to investigate the association between host factors and AMR. Overall, 553/2,738 (20.2%) of the isolates were susceptible to 17 of the 27 antimicrobials examined. Of the 2,638 isolates examined for amikacin susceptibility, 2,706 (97.5%) were susceptible, 2,657/2,673 (99.4%) isolates were susceptible to imipenem, and 2,099/2,670 (78.6%) were susceptible to marbofloxacin. A significant decreasing trend in susceptibility was observed for amoxicillin-clavulanic acid (P<0.0001), ampicillin (P<0.0001), Cefazolin (P<0.0001), ceftazidime (P = 0.0067), chloramphenicol (P<0.0001), and orbifloxacin (P = 0.008). The overall percentage of AMR isolates (isolates not susceptible to at least one antimicrobial) was 61.7% (1,690/2,738) and 29.3% (801/2,738) of isolates were multidrug resistant. Multivariable regression analyses showed significant associations between AMR and age (P = 0.0091), breed (P = 0.0008), and sample isolation site/source (P<0.0001). The decreasing trend in the proportion of isolates susceptible to several beta-lactam antimicrobials suggests that resistance of Escherichia coli in dogs to these antimicrobials could be increasing in Indiana. The decreasing trend in susceptibility to these drugs could be due to selection pressure from antimicrobial use.

Presumed acquired dynamic pectus excavatum in a cat.

An 11-year-old, spayed female American domestic shorthair cat was presented with open-mouth breathing and dyspnea due to respiratory tract disease. Radiographically, there was thickening of the retropharyngeal soft tissue and soft palate, focal pulmonary change, and severe pectus excavatum (PE), best seen on the left lateral view. Repeat thoracic radiographs 4 months later showed persistent PE, but only on the right lateral view; PE was absent on the left lateral view. The dynamic component of the observed PE was presumably acquired secondary to chronic upper respiratory disease and/or concurrent increased pliability of the thoracic wall from prolonged prednisolone treatment.

Pectus excavatum dynamique acquis présumé chez un chat. Une chatte domestique américaine stérilisée de 11 ans a présenté une respiration à bouche ouverte et une dyspnée due à une maladie des voies respiratoires. Radiographiquement, il y avait un épaississement des tissus mous rétropharyngés et du palais mou, une modification pulmonaire focale et un pectus excavatum (PE) sévère, mieux visible sur la vue latérale gauche. Des radiographies thoraciques répétées 4 mois plus tard ont montré une PE persistante, mais uniquement sur la vue latérale droite; la PE était absent sur la vue latérale gauche. La composante dynamique de la PE observée était vraisemblablement acquise secondaire à une maladie chronique des voies respiratoires supérieures et/ou à une flexibilité accrue concomitante de la paroi thoracique à la suite d'un traitement prolongé à la prednisolone.(Traduit par Dr Serge Messier).

Comparison of 2 assays for measuring serum total thyroxine concentration in dogs and cats.

BACKGROUND: No gold standard assay for serum total thyroxine (TT4) concentration in small animals exists. The Microgenics DRI TT4 (MTT4) assay is used by most reference laboratories. HYPOTHESIS/OBJECTIVES: IDEXX Catalyst Total T4 (CTT4) and Immulite 2000 TT4 (ITT4) results will agree with MTT4 results. ANIMALS: Residual small animal sera were randomized before reanalysis (dogs, CTT4 versus MTT4: n = 176, ITT4 versus MTT4: n = 74; cats, CTT4 versus MTT4: n = 319, ITT4 versus MTT4: n = 79). METHODS: Validation and method comparison study. Serum TT4 concentration was measured on all analyzers. Pairwise Pearson correlation, cumulative sum linearity test, regression, and Bland-Altman method were performed. RESULTS: CTT4 versus MTT4 in dogs: constant bias (y-intercept) was 0.10 µg/dL (95% confidence interval [CI], 0.05-0.15), proportional bias (slope) was 0.86 µg/dL (95% CI, 0.83-0.89); in cats, constant bias was 0.13 µg/dL (95% CI, 0.08-0.20) and proportional bias was 1.01 µg/dL (95% CI, 0.98-1.03), but the test for linearity failed. Bland-Altman plots identified increasing disagreement with increasing serum TT4 concentrations. ITT4 versus MTT4 in dogs, constant bias was 0.14 µg/dL (95% CI, 0.04-0.22) and 0.22 µg/dL (95% CI, 0.09-0.33) for cats; proportional bias was 0.76 (95% CI, 0.72-0.80) for dogs and 0.71 (95% CI, 0.69-0.74) for cats. CONCLUSIONS AND CLINICAL IMPORTANCE: Differences in CTT4 and MTT4 results affect interpretation at higher serum TT4 concentrations. The ITT4 proportional bias will underestimate serum TT4 concentrations in dogs and cats, compared to MTT4. Serial TT4 measurements should be done using the same assay.

Serologic investigation of exposure to influenza A virus H3N2 infection in dogs and cats in the United States.

We investigated, in a cross-sectional study, the prevalence of antibodies against canine influenza A virus (CIV) H3N2 in serum samples collected from dogs and cats using a commercial ELISA and a hemagglutination inhibition (HI) test. Samples were obtained from 519 cats and dogs from 13 states within the United States. Data were analyzed for potential risk factors with positive sera (vs. negative sera) by logistic regression. Odds ratios and their 95% confidence intervals (CIs) were calculated by exponentiation of the regression coefficients. Ten dogs (2.21%; 95% CI: 1.05-3.98%) and 6 cats (8.96%; 95% CI: 3.36-18.48%) tested seropositive for CIV H3N2 by HI. One feline sample (1.49%; 95% CI: 0.04-8.04%) and 16 canine samples (3.53%; 95% CI: 2.01-5.61%) tested seropositive by ELISA for influenza A virus. There were no apparent associations between seropositivity and putative risk factors. All positive animals were from Indiana or Illinois; however, CIV H3N2 seroprevalence was not common in Illinois and Indiana.

Patient management.

Hospital-associated infections, including those caused by zoonotic agents, represent an increasing concern in veterinary practice. Veterinarians and hospital staff are obligated and expected to provide education about and protection from transmission of pathogens among animal patients and between animal patients and human beings (eg, veterinary staff, volunteers, owners) who come into contact with infected animals. Patient management involves assessing risks of pathogen transmission, identification of animals either suspected of or proved to be infected with a transmissible infectious disease agent, and the implementation of measures that minimize the likelihood of transmission of the infectious agent.

Prevalence of methicillin-resistant Staphylococcus spp. in the conjunctival sac of healthy dogs.

OBJECTIVES: To determine the prevalence of selected coagulase-positive methicillin-resistant Staphylococcus aureus (MRS) in the conjunctival sac in a group of healthy dogs and to compare the prevalence of ocular MRS colonization with colonization of typically assessed body sites including the nasal cavity and rectum. ANIMALS STUDIED: 123 healthy dogs were used in the prevalence study: 40 dogs from a shelter and 83 privately owned dogs. PROCEDURES: The sampling procedure included culturing three separate sites per subject in the following order: the lower conjunctival fornices, the nares, and rectum. RESULTS: A low prevalence of 1.6% (2/123) of MRS was detected in healthy dogs. Methicillin-resistant Staphylococcus pseudintermedius was isolated from two dogs, one from a conjunctival swab and the other from a rectal swab. CONCLUSION: The survey data indicate the ocular surface is a potential site of MRS colonization, although the prevalence was low in healthy dogs.

In vitro effect of multiple hydrogen peroxide gas plasma sterilizations on the rate of closure of ameroid constrictors.

OBJECTIVE: To evaluate the effect of multiple hydrogen peroxide gas plasma (HPGP) sterilizations on the rate of closure of ameroid constrictors. SAMPLE: Thirty-six 5.0-mm ameroid constrictors. PROCEDURES: Ameroid constrictors were randomly allocated to 6 groups. Each group underwent 1, 2, 3, 4, 5, or 6 HPGP sterilizations. Ameroid constrictors were then incubated for 35 days in canine plasma and digitally imaged at predetermined times during incubation. One individual, who was unaware of the group to which each ameroid constrictor was assigned, measured the lumen area of the constrictor on each digital image. Mean lumen area was compared among groups. RESULTS: No ameroid constrictors were completely closed after 35 days of incubation in canine plasma. Mean lumen area after incubation did not differ among constrictors that underwent 1, 2, and 3 sterilizations. Constrictors that underwent 4 sterilizations were closed significantly more than were those that underwent 1, 2, or 3 sterilizations. Mean lumen area after incubation did not differ significantly between constrictors that underwent 5 and 6 sterilizations, although the final lumen areas for those constrictors were significantly smaller than those for constrictors that underwent 1, 2, 3, and 4 sterilizations. CONCLUSIONS AND CLINICAL RELEVANCE: Ameroid constrictors that underwent 5 and 6 HPGP sterilizations had a 9% to 12% decrease in lumen area, compared with that of constrictors that underwent ≤ 4 plasma sterilizations, and the use of such constrictors could increase the risk of portal hypertension and secondary acquired shunting or decrease the risk of persistent shunting.

Antibacterial activity of novel cationic peptides against clinical isolates of multi-drug resistant Staphylococcus pseudintermedius from infected dogs.

Staphylococcus pseudintermedius is a major cause of skin and soft tissue infections in companion animals and has zoonotic potential. Additionally, methicillin-resistant S. pseudintermedius (MRSP) has emerged with resistance to virtually all classes of antimicrobials. Thus, novel treatment options with new modes of action are required. Here, we investigated the antimicrobial activity of six synthetic short peptides against clinical isolates of methicillin-susceptible and MRSP isolated from infected dogs. All six peptides demonstrated potent anti-staphylococcal activity regardless of existing resistance phenotype. The most effective peptides were RRIKA (with modified C terminus to increase amphipathicity and hydrophobicity) and WR-12 (α-helical peptide consisting exclusively of arginine and tryptophan) with minimum inhibitory concentration50 (MIC50) of 1 µM and MIC90 of 2 µM. RR (short anti-inflammatory peptide) and IK8 "D isoform" demonstrated good antimicrobial activity with MIC50 of 4 µM and MIC90 of 8 µM. Penetratin and (KFF)3K (two cell penetrating peptides) were the least effective with MIC50 of 8 µM and MIC90 of 16 µM. Killing kinetics revealed a major advantage of peptides over conventional antibiotics, demonstrating potent bactericidal activity within minutes. Studies with propidium iodide and transmission electron microscopy revealed that peptides damaged the bacterial membrane leading to leakage of cytoplasmic contents and consequently, cell death. A potent synergistic increase in the antibacterial effect of the cell penetrating peptide (KFF)3K was noticed when combined with other peptides and with antibiotics. In addition, all peptides displayed synergistic interactions when combined together. Furthermore, peptides demonstrated good therapeutic indices with minimal toxicity toward mammalian cells. Resistance to peptides did not evolve after 10 passages of S. pseudintermedius at sub-inhibitory concentration. However, the MICs of amikacin and ciprofloxacin increased 32 and 8 fold, respectively; under similar conditions. Taken together, these results support designing of peptide-based therapeutics for combating MRSP infections, particularly for topical application.

Seroprevalence of Ehrlichia canis, Ehrlichia chaffeensis and Ehrlichia ewingii in dogs in North America.

BACKGROUND: This study evaluated the exposure of dogs to three different Ehrlichia spp. in the south and central regions of the United States where vector-borne disease prevalence has been previously difficult to ascertain, particularly beyond the metropolitan areas. METHODS: Dog blood samples (n = 8,662) were submitted from 14 veterinary colleges, 6 private veterinary practices and 4 diagnostic laboratories across this region. Samples were tested for E. canis, E. chaffeensis and E. ewingii specific antibodies using peptide microtiter ELISAs. RESULTS: Overall, E. canis, E. chaffeensis and E. ewingii seroprevalence was 0.8%, 2.8%, and 5.1%, respectively. The highest E. canis seroprevalence (2.3%) was found in a region encompassing Arkansas, Louisiana, Oklahoma, Tennessee and Texas. E. chaffeensis seroreactivity was 6.6% in the central region (Arkansas, Kansas, Missouri, and Oklahoma) and 4.6% in the southeast region (Georgia, Maryland, North Carolina, South Carolina, Tennessee and Virginia). Seroreactivity to E. ewingii was also highest in the central region (14.6%) followed by the southeast region (5.9%). The geospatial pattern derived from E. chaffeensis and E. ewingii seropositive samples was similar to previous reports based on E. chaffeensis seroreactivity in white-tailed deer and the distribution of human monocytic ehrlichiosis (HME) cases reported by the CDC. CONCLUSIONS: The results of this study provide the first large scale regional documentation of exposure to E. canis, E. chaffeensis and E. ewingii in pet dogs, highlighting regional differences in seroprevalence and providing the basis for heightened awareness of these emerging vector-borne pathogens by veterinarians and public health agencies.

Feline bartonellosis.

Bartonella infection is common among domestic cats, but the role of Bartonella species as feline pathogens requires further study. Most Bartonella species that infect cats are zoonotic. Cats are the mammalian reservoir and vector for Bartonella henselae, an important zoonotic agent. Cat fleas transmit Bartonella among cats, and cats with fleas are an important source of human B henselae infections. New information about Bartonella as feline pathogens has recently been published, and this article summarizes much of that information. Issues surrounding diagnosis and treatment of feline Bartonella infections are described, and prevention of zoonotic transmission of Bartonella is discussed.

Detection of antibodies against Leptospira serovars via microscopic agglutination tests in dogs in the United States, 2000-2007.

OBJECTIVE: To use results of microscopic agglutination tests (MATs) conducted at a commercial veterinary diagnostic laboratory to determine temporal and demographic distributions of positive serologic test results for leptospirosis in dogs and identify correlations among results for various Leptospira serovars. DESIGN: Serosurvey. STUDY POPULATION: MAT results for 33,119 canine serum samples submitted to a commercial veterinary diagnostic laboratory from 2000 through 2007. PROCEDURES: Electronic records of MAT results for dogs were obtained from a veterinary diagnostic laboratory. Seropositivity for antibodies against Leptospira serovars was determined by use of a cutoff titer of >or=1:1,600 to reduce the possible impact of postvaccinal antibodies on results. Correlations between results for all possible pairs of serovars were calculated by ordinal ranking of positive (>or=1:100) antibody titer results. RESULTS: 2,680 samples (8.1%; 95% confidence interval [CI], 7.8% to 8.4%) were seropositive for antibodies against Leptospira serovars. The highest percentage of positive MAT results was for the year 2007 (10.2%; 95% CI, 9.5% to 10.9%) and for the months of November and December during the study period. Antibodies were most common against serovars Autumnalis, Grippotyphosa, Pomona, and Bratislava. Seroprevalence of leptospirosis was lowest for dogs>10 years of age but was similar across other age strata. CONCLUSIONS AND CLINICAL RELEVANCE: Leptospirosis can affect dogs of small and large breeds and various ages. Although an increase in proportions of positive MAT results was evident in the fall, monthly and annual variations suggested potential exposure in all months. Because of the limitations of MAT results and the limited number of serovars used in the test, bacterial culture should be used to identify infective Leptospira serovars.

Spatial and spatio-temporal clustering of overall and serovar-specific Leptospira microscopic agglutination test (MAT) seropositivity among dogs in the United States from 2000 through 2007.

Leptospirosis is a re-emerging disease of dogs in the United States (U.S.). This paper reports the findings of a retrospective study conducted to determine if seroreactivity to Leptospira microscopic agglutination test (MAT) among dogs in the U.S. clustered in space and time. The study utilized canine sera submitted to a commercial laboratory for leptospiral MAT from January 2000 through December 2007. There were 31,869 serum samples submitted by veterinarians from 3156 zip code locations across the U.S. Results of MAT were considered positive at titers of > or = 1:1600. Spatial and spatial-temporal scan statistics were used to identify statistically significant clusters of seroreactivity to Leptospira (overall and individual serovars) using recorded test request dates and locations of the centroid of the zip code reported for each serum sample. There were 2469 positive MAT results with a titer > or = 1:1600 to at least one of seven Leptospira serovars. Two relevant spatial clusters of 26.3 and 246.5 km radius were identified (P=0.001). The primary cluster was located in the northeastern part of Illinois including Chicago and surrounding areas (232 [14.4%] of 1612 MAT positive; RR=1.95). The secondary cluster covered the central part of Texas (292 [12.62%] of 2314 MAT positive; RR=1.71). Eight space-time clusters of overall MAT positivity were identified (29-335 km radius; P=0.001-0.048 and RR=3.98-24.69) that covered different geographic locations for different time points. Spatial and space-time clusters for individual serovars were also identified for six serovars: eight each of Grippotyphosa and Pomona, seven of Bratislava, five of Autumnalis, and three each of Icterohaemorrhagiae and Canicola. In conclusion Leptospira seropositivity in dogs tended to have distinctive clusters in space and space-time. Most of the space-time clusters of overall Leptospira MAT seropositivity were associated with cluster events for individual serovars. Further investigation is warranted to explain individual serovar clusters detected in this study, as a complex interaction of incidental host, environment and reservoir host may be responsible for the occurrence of these serovar clusters.

Bartonellosis.

Bartonellosis is a constellation of clinical conditions affecting human beings and a variety of animals. Many Bartonella infections are zoonotic, with some of the most commonly reported zoonotic manifestations of infection including cat scratch disease, bacillary angiomatosis, endocarditis, and neuroretinitis. Companion animals serve as reservoirs for several zoonotic species of Bartonella, and may also serve as sentinels for zoonotic Bartonella species harbored by wildlife. This article provides an overview of bartonellosis of dogs and cats, and discusses public health implications of animal bartonellosis.

Failure to identify an association between serologic or molecular evidence of Bartonella infection and idiopathic rhinitis in dogs.

OBJECTIVE: To determine whether infection with or exposure to Bartonella spp was associated with idiopathic rhinitis in dogs. DESIGN: Case-control study. ANIMALS: 44 dogs with idiopathic nasal discharge and 63 age- and weight-matched control dogs without nasal discharge and no clinical signs of bartonellosis. Procedures-Serum was tested for antibodies against Bartonella henselae and Bartonella vinsonii subsp berkhoffii with indirect fluorescent antibody assays. Blood was tested for Bartonella DNA with a PCR assay. RESULTS: Results of the antibody and PCR assays were negative for all 44 dogs with idiopathic nasal discharge. One control dog had antibodies against B henselae; a second control dog had positive PCR assay results. We did not detect a significant association between assay results and group designation. CONCLUSIONS AND CLINICAL RELEVANCE: The present study failed to confirm an association between idiopathic rhinitis and exposure to or infection with Bartonella spp in dogs. Findings do not rule out the possibility that Bartonella infection may cause nasal discharge in some dogs, but the failure to find any evidence of exposure to or infection with Bartonella spp in dogs with idiopathic nasal discharge suggested that Bartonella infection was not a common cause of the disease.

Adverse events after vaccine administration in cats: 2,560 cases (2002-2005).

OBJECTIVE: To determine the incidence of vaccine-associated adverse events (VAAEs) diagnosed within 30 days of vaccination in cats and characterize risk factors for their occurrence. DESIGN: Retrospective cohort study. ANIMALS: 496,189 cats vaccinated at 329 hospitals. PROCEDURES: Electronic records were searched for VAAEs that occurred after vaccine administration classified by practitioners as nonspecific vaccine reaction, allergic reaction, urticaria, shock, or anaphylaxis. Clinical signs and treatments were reviewed. The association between potential risk factors and a VAAE occurrence was estimated via multivariate logistic regression. RESULTS: 2,560 VAAEs were associated with administration of 1,258,712 doses of vaccine to 496,189 cats (51.6 VAAEs/10,000 cats vaccinated). The risk of a VAAE significantly increased as the number of vaccines administered per office visit increased. Risk was greatest for cats approximately 1 year old; overall risk was greater for neutered versus sexually intact cats. Lethargy with or without fever was the most commonly diagnosed VAAE. No localized reactions recorded in the 30-day period were subsequently diagnosed as neoplasia when followed for 1 to 2 years. CONCLUSIONS AND CLINICAL RELEVANCE: Although overall VAAE rates were low, young adult neutered cats that received multiple vaccines per office visit were at the greatest risk of a VAAE within 30 days after vaccination. Veterinarians should incorporate these findings into risk communications and limit the number of vaccinations administered concurrently to cats.

Bartonella henselae exists as a mosaic of different genetic variants in the infected host.

Bartonella henselae is a fastidious bacterium associated with infections in humans and cats. The mechanisms involved in the long-term survival of bartonellae despite vigorous host immune responses are poorly understood. Generation of genetic variants is a possible strategy to circumvent the host specific immune responses. The authors have recently demonstrated the coexistence of different genetic variants within the progeny of three primary B. henselae isolates from Berlin by PFGE analysis. Aims of the present study were to determine whether coexistence of different variants is a common feature of B. henselae isolates worldwide and whether the genetic variants originally emerged in vivo. Thirty-four primary isolates from different geographical regions were analysed by subjecting multiple single-colony-derived cultures to PFGE analysis. Up to three genetic variants were detected within 20 (58.8 %) isolates, indicating that most primary isolates display a mosaic-like structure. The close relatedness of the genetic variants within an isolate was confirmed by multi-locus sequence typing. In contrast to the primary isolates, no genetic variants were detected within the progeny of 20 experimental clones generated in vitro from 20 primary isolates, suggesting that the variants were not induced in vitro during the procedure of PFGE analysis. Hence, the genetic variants within a primary isolate most likely originally emerged in vivo. Consideration of the mosaic structure of primary isolates is essential when interpreting typing studies on B. henselae.