Bacterial Infections In Dogs Research Articles

Genomic Analyses of Methicillin-Susceptible and Methicillin-Resistant Staphylococcus pseudintermedius Strains Involved in Canine Infections: A Comprehensive Genotypic Characterization.

Staphylococcus pseudintermedius is frequently associated with several bacterial infections in dogs, highlighting a One Health concern due to the zoonotic potential. Given the clinical significance of this pathogen, we performed comprehensive genomic analyses of 28 S. pseudintermedius strains isolated from canine infections throughout whole-genome sequencing using Illumina HiSeq, and compared the genetic features between S. pseudintermedius methicillin-resistant (MRSP) and methicillin-susceptible (MSSP) strains. Our analyses determined that MRSP genomes are larger than MSSP strains, with significant changes in antimicrobial resistance genes and virulent markers, suggesting differences in the pathogenicity of MRSP and MSSP strains. In addition, the pangenome analysis of S. pseudintermedius from canine and human origins identified core and accessory genomes with 1847 and 3037 genes, respectively, which indicates that most of the S. pseudintermedius genome is highly variable. Furthermore, phylogenomic analysis clearly separated MRSP from MSSP strains, despite their infection sites, showing phylogenetic differences according to methicillin susceptibility. Altogether our findings underscore the importance of studying the evolutionary dynamics of S. pseudintermedius, which is crucial for the development of effective prevention and control strategies of resistant S. pseudintermedius infections.

Assessing Illinois companion animal veterinarians' antimicrobial prescription practices and the factors that influence their decisions when treating bacterial infections in dogs and cats.

Judicious antimicrobial use in companion animal practice is critical for maintaining the effectiveness of antimicrobial agents against bacterial infections and reducing the selection of antimicrobial-resistant bacteria. This study aimed to provide insights into companion animal veterinarians' antimicrobial treatment recommendations for common bacterial infections in dogs and cats and describe the factors influencing their prescription choices. An online survey using QualtricsXM® software was administered between September and November 2022 to companion animal veterinarians who were Illinois State Veterinary Medical Association members. Descriptive and text analyses were conducted to assess the participants' responses. A total of 78 surveys were included in the analysis. Skin infections were ranked as the most common bacterial infections for which veterinarians prescribed antimicrobial agents, followed by ear, urinary tract, respiratory, and enteric infections. The severity of clinical symptoms and the results of bacterial culture and susceptibility tests were the most influential factors for veterinarians when making antimicrobial prescription choices. Veterinarians were aware of the current antimicrobial prescription guideline recommendations when prescribing antimicrobials empirically to nine hypothetical scenarios of bacterial infections. According to the results of the text analysis that assessed veterinarians' responses to an open-ended question, regarding their challenges when prescribing antimicrobial agents, the pairwise correlation of word frequencies within each response showed the highest correlations between the words 'owner' and 'compliance', 'administration' and 'route', 'cost' and 'culture', and 'patients' and 'acceptance'. The study results can support animal health stakeholders in the development of antimicrobial stewardship programmes to promote appropriate antimicrobial use and limit the emergence of antimicrobial resistance.

Prospective comparison of prostatic aspirate culture and cystocentesis urine culture for detection of bacterial infection in dogs with prostatic neoplasia.

The purpose of this study was to determine whether prostatic aspirate culture is a superior method to detect infection compared to culture of urine collected by cystocentesis in dogs with prostatic neoplasia. A prospective study was conducted and dogs with suspected or confirmed prostatic neoplasia were enrolled. Urinalysis was done and culture and antimicrobial susceptibility testing was performed on paired urine and prostatic aspirate samples collected at a single timepoint. Ten dogs with prostatic neoplasia were enrolled. All dogs had one or more clinical sign consistent with lower urinary tract disease. One dog (10%) had a positive urine culture, but negative prostatic aspirate culture, one dog (10%) had a positive prostatic aspirate culture, but negative urine culture, and one dog (10%) had both positive urine and prostatic aspirate cultures. Using prostatic aspirate culture as the reference standard, urine culture had a sensitivity for detecting infection of 87.5% (95% confidence interval 52.9 to 99.4) and specificity of 50% (92.6 to 97.4) in this population of dogs. Positive cultures were uncommon with both culture collection methods. Study results did not identify prostatic aspirate culture to be a more sensitive method of detecting prostatic infection than urine culture collected by cystocentesis in these dogs with prostatic neoplasia.

Activated Mesenchymal Stromal Cell Therapy for Treatment of Multi-Drug Resistant Bacterial Infections in Dogs.

New and creative approaches are required to treat chronic infections caused by increasingly drug-resistant strains of bacteria. One strategy is the use of cellular therapy employing mesenchymal stromal cells (MSC) to kill bacteria directly and to also activate effective host immunity to infection. We demonstrated previously that activated MSC delivered systemically could be used effectively together with antibiotic therapy to clear chronic biofilm infections in rodent models. Therefore, we sought in the current studies to gain new insights into the antimicrobial properties of activated canine MSC and to evaluate their effectiveness as a novel cellular therapy for treatment of naturally-occurring drug resistant infections in dogs. These studies revealed that canine MSC produce and secrete antimicrobial peptides that synergize with most classes of common antibiotics to trigger rapid bactericidal activity. In addition, activated canine MSC migrated more efficiently to inflammatory stimuli, and secreted factors associated with wound healing and fibroblast proliferation and recruitment of activated neutrophils. Macrophages incubated with conditioned medium from activated MSC developed significantly enhanced bactericidal activity. Clinical studies in dogs with chronic multidrug resistant infections treated by repeated i.v. delivery of activated, allogeneic MSC demonstrated significant clinical benefit, including infection clearance and healing of infected tissues. Taken together, the results of these studies provide new insights into antimicrobial activity of canine MSC, and their potential clinical utility for management of chronic, drug-resistant infections.

Bacterial infection in dogs with aspiration pneumonia at 2 tertiary referral practices.

BackgroundIn dogs, antimicrobial drugs are widely prescribed for aspiration pneumonia (AP) despite poor documentation of bacterial infection in AP (b‐AP) using bronchoalveolar lavage fluid (BALF) analysis. Interpretating discordant cytology and culture results is challenging, contributing to lack of a criterion standard, and highlighting differences between veterinary and human medical criteria for b‐AP.ObjectivesDetermine how many dogs with AP had BALF collection and differences in diagnosis of b‐AP using veterinary vs human medical criteria. Report findings of noninvasive markers (e.g. fever, band neutrophilia, radiographic severity score) in dogs with and without b‐AP.AnimalsRetrospective cohort study of client‐owned dogs (n = 429) with AP at 2 university veterinary hospitals. Twenty‐four dogs met enrollment criteria.MethodsInclusion criteria were radiographic diagnosis of AP, ≥1 risk factor, CBC findings, and BALF cytology and culture results. Veterinary medical b‐AP criteria were cytology findings compatible with sepsis with or without positive culture, or cytology findings not consistent with sepsis and positive culture (≥1.7 × 103 cfu/mL). Human medical b‐AP criteria required culture with ≥104 cfu/mL or > 7% cells with intracellular bacteria on cytology.ResultsOnly 24/429 dogs met all enrollment criteria; 379/429 dogs lacked BALF collection. Diagnosis of b‐AP differed using veterinary (79%) vs human (29%) medical criteria. Fever, band neutrophils and high radiographic scores were noted in dogs with and without b‐AP.Conclusions and Clinical ImportanceLack of routine BALF collection hampers definitive recognition of bacterial infection in AP. Differences in dogs meeting veterinary vs human medical definitions for b‐AP and usefulness of noninvasive markers warrant further study to improve understanding of the role of bacteria in AP.

Antibacterial Properties of Canine Platelet-Rich Plasma and Other Non-Transfusional Hemo-Components: An in vitro Study.

This in vitro study was carried out to evaluate the potential antibacterial properties of canine non-transfusional hemo-components. Therapeutic formulations commonly used for regenerative medicine purposes (platelet-rich plasma, platelet gel, platelet lysate, fibrin glue), considering both leukocyte-rich and leukocyte-poor formulations, but also platelet-poor plasma and activating substances (thrombin, calcium gluconate), were tested to detect elements with potential antimicrobial properties. The antibacterial effect was tested on different bacterial strains (Staphylococcus aureus subspecies aureus, Staphylococcus cohnii subspecies cohnii, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae subspecies pneumoniae) isolated from canine wounds and classified as susceptible, multidrug-, extensively, and pandrug-resistant bacteria toward a known panel of human and veterinary antibiotics. The evaluation was carried out by agar gel diffusion method (Kirby–Bauer) and micro-inhibition in broth using microplates and spectrophotometer reading. The study findings confirmed the hypothesized antibacterial properties of canine non-transfusional hemo-components. A more effective bacteriostatic effect was found against Gram-negative bacteria, drug-resistant too. The presence of leukocytes or platelets does not appear to be essential for the antibacterial effect. Further studies should be conducted to evaluate the exact mechanism of action of the antimicrobial activity. However, non-transfusional hemo-components could be a useful natural aid in controlling bacterial infections in dogs.

Bacterial infection before and after stent placement in dogs with tracheal collapse syndrome.

BackgroundDogs with tracheal stents often have positive airway bacterial cultures. The pathogenicity of these organisms and risk factors for infection have not been investigated.ObjectiveDescribe bacterial infection in dogs with tracheal collapse before and after tracheal stent placement.AnimalsFifty‐three client‐owned dogs.MethodsRetrospective review of medical records of dogs receiving tracheal stents with thoracic radiographs, tracheoscopy, and endotracheal lavage.ResultsThere was no difference between the overall prevalence of dogs with positive bacterial cultures before (31/38; 82%) or after stent placement (24/31; 77%) (P = .67). An increased number of geriatric (17/28; 61%) and traditional‐type collapse (TTC) (16/26; 62%) dogs had positive pathogenic airway infections before stent placement, compared to young (8/25; 32%; P = .04) and malformation‐type collapse (MTC) dogs (9/27; 33%; P = .04). After tracheal stent placement, geriatric dogs had a 52% reduction in pathogenic bacteria infection frequency (P = .02) and dogs with TTC had a 56% reduction in pathogenic bacteria infection frequency (P = .01). Significant risk factors for pathogenic infection included a history of pneumonia (OR = 3.6; 95% CI, 0.28‐43.36) and cardiac disease (OR = 1.25; 95% CI, 0.16‐9.92) in geriatric dogs, and hepatomegaly in young dogs (OR = 1.5; 95% CI, 0.12‐19.44).Conclusions and Clinical ImportanceTracheal stent placement does not increase the overall rate of pathogenic bacterial infection in dogs with tracheal collapse and can decrease the rate of subsequent pathogenic infections in geriatric dogs and dogs with TTC that require tracheal stenting. Airway culture and cytology should be performed in all dogs undergoing tracheal stent placement.

Local immune and microbiological responses to mucosal administration of a Liposome-TLR agonist immunotherapeutic in dogs

BackgroundNon-specific immunotherapeutics have been evaluated previously in dogs, primarily for cancer treatment. However, there remains a need for a more broadly targeted, general purpose immunotherapeutic capable of activating innate immune defenses for non-specific protection or early treatment of viral and bacterial infections. To address need, our group has developed a liposomal immune stimulant (liposome-TLR complexes, LTC) containing TLR 3 and 9 agonists specifically designed to activate mucosal immune defenses in sites such as nasal cavity and oropharynx, following topical delivery. In this study, we evaluated the local immune stimulatory properties of LTC in vitro and in healthy purpose-bred dogs, including activation of cellular recruitment and cytokine production. The ability of LTC treatment to elicit effective antiviral immunity was assessed in dogs following a canine herpesvirus outbreak, and the impact of LTC treatment on the local microbiome of the oropharynx was also investigated.ResultsThese studies revealed that LTC potently activated innate immune responses in vitro and triggered significant recruitment of inflammatory monocytes and T cells into the nasal cavity and oropharynx of healthy dogs. Administration of LTC to dogs shortly after an outbreak of canine herpesvirus infection resulted in significant reduction in clinical signs of infection. Interestingly, administration of LTC to healthy dogs did not disrupt the microbiome in the oropharynx, suggesting resiliency of the microflora to transient immune activation.ConclusionsTaken together, these results indicate that LTC administration mucosally to dogs can trigger local innate immune activation and activation of antiviral immunity, without significantly disrupting the composition of the local microbiome. Thus, the LTC immune stimulant has potential for use as a non-specific immunotherapy for prevention or early treatment of viral and bacterial infections in dogs.

Staphylococcal pyoderma: challenges and therapeutic considerations

Superficial staphylococcal pyoderma is a common diagnosis in companion animal practice. The majority of cutaneous bacterial infections in dogs and cats are associated with Staphylococcus pseudintermedius , Staphylococcus aureus or...

Serological and molecular detection of spotted fever group Rickettsia in a group of pet dogs from Luanda, Angola

BackgroundInfections with tick-borne rickettsiae can cause diseases well known in humans but still not so well characterized in dogs. Susceptibility to infection depends on the virulence of Rickettsia spp. and only a few of them have been described to cause disease in dogs. The aim of this study was to investigate the exposure to Rickettsia spp. among a group of pet dogs from Luanda, Angola.ResultsOut of 103 dogs included in the study, 62 (60.2%) were infested with ticks. Plasma specimens tested for serology by an immunofluorescence assay (IFA) revealed that six (5.8%) dogs had detectable immunoglobulin G (IgG) antibodies to spotted fever group Rickettsia (SFGR), with endpoint titers of 64 for two dogs, 128 for three dogs and 1024 for one dog. From the seropositive group of dogs, five (83%) of them were males, with their age ranging from 1 to 8 years old. Among the seropositive dogs, four (66.7%) were parasitized with ticks and no breed (or cross) was found to be associated with specific antibodies. Rickettsia spp. DNA was detected by nested-polymerase chain reaction (PCR) in two (1.9%) dogs that were found to be seronegative.ConclusionsSeroprevalence and molecular detection of Rickettsia spp. infection in this group of pet dogs from Luanda is low compared with other studies performed in the same type of hosts in other areas. Although many dogs were parasitized with ticks, a low prevalence of Rickettsia spp. could be related with the hypothesis of a low rickettsial prevalence in the infesting ticks. This study provides evidence that dogs in Luanda are exposed to Rickettsia spp., but further studies are needed to better characterize the bacterial infections in dogs and in their ectoparasites.

Antimicrobial use Guidelines for Treatment of Respiratory Tract Disease in Dogs and Cats: Antimicrobial Guidelines Working Group of the International Society for Companion Animal Infectious Diseases.

Respiratory tract disease can be associated with primary or secondary bacterial infections in dogs and cats and is a common reason for use and potential misuse, improper use, and overuse of antimicrobials. There is a lack of comprehensive treatment guidelines such as those that are available for human medicine. Accordingly, the International Society for Companion Animal Infectious Diseases convened a Working Group of clinical microbiologists, pharmacologists, and internists to share experiences, examine scientific data, review clinical trials, and develop these guidelines to assist veterinarians in making antimicrobial treatment choices for use in the management of bacterial respiratory diseases in dogs and cats.

Cefovecin Pharmacokinetics in the Red-Eared Slider

The administration of long-acting antimicrobial agents to treat susceptible bacterial infections in exotic animals is desirable, as it may reduce handling stress, increase therapeutic compliance, and decrease morbidity and mortality. Cefovecin sodium, a third-generation cephalosporin, is administered to treat susceptible bacterial infections in dogs and cats. Cefovecin has an extended dosing interval partially because of increased ability to bind with plasma protein. As such, investigations into the pharmacokinetics of this antibiotic have been conducted in zoo and exotic animal species, including terrestrial chelonians. The objective of this project was to determine the pharmacokinetic profile and protein binding of cefovecin in the red-eared slider (Trachemys scripta elegans). Eleven healthy red-eared sliders were administered 10mg/kg of cefovecin subcutaneously in the forelimb. Blood samples were obtained at 2, 4, 8, 12, 24, and 48 hours postadministration. Plasma cefovecin concentrations were analyzed by reversed-phase high-performance liquid chromatography, and protein binding was determined using ultrafiltration. Noncompartmental pharmacokinetic analysis was performed on the resulting data. Cefovecin was rapidly absorbed, with a maximal measured plasma drug concentration of 32.3 (±23.8)µg/mL occurring at 2.4 (±1.2) hours and a terminal-phase half-life of 6.8 (±1.2) hours, which was much shorter than the half-life observed in dogs and cats (approximately 5 and 7 days, respectively). No substantial side effects were noted in subjects following antibiotic administration. Although well tolerated in red-eared sliders, the rapid decline in plasma cefovecin concentration, presumably due to the absence of plasma protein binding, negates its use as a long-acting antibiotic in this species.

Multiplex cytokine analyses in dogs with pyometra suggest involvement of KC-like chemokine in canine bacterial sepsis

Clinical diagnostic criteria for sepsis (systemic inflammatory response syndrome caused by infection) are unspecific and, therefore, biomarkers for sepsis diagnosis are needed for appropriate treatment and patient survival. Pyometra, a common disease caused by bacterial infection of the uterus, results in sepsis in nearly 60% of cases in dogs. We used dogs with pyometra as a natural model for sepsis and collected serum samples from 39 dogs, of which 22 with pyometra and 17 healthy controls. Dogs with pyometra were further grouped into dogs with sepsis (n=18) and without sepsis (n=4). Serum concentrations of a panel of cytokines, including keratinocyte-derived chemokine (KC)-like, granulocyte-macrophages colony stimulating factor (GM-CSF), interleukin (IL)-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18, chemokine C-X-C motif ligand (CXCL)10 and tumor necrosis factor (TNF)-α, were measured using multiplex analyses. Serum C-reactive protein (CRP) levels were determined using an automated immunoturbidimetric assay. In addition to physical examination hematological and serum biochemical analyses were performed to evaluate the overall status of the dogs. Significantly higher concentrations of KC-like (757 vs 304pg/ml) were detected in dogs with pyometra as compared to healthy dogs. Within the pyometra group, dogs with sepsis compared to dogs without sepsis had a higher KC-like concentration (873 vs 300pg/ml). Hemoglobin levels were significantly lower in dogs with pyometra compared to healthy dogs, regardless of the presence or absence of sepsis, and correlated negatively with KC-like. KC-like concentrations correlated positively with CRP, number of hospitalization days, number of monocytes, concentrations of IL-8, and percentage band neutrophils. Our data suggest that bacterial infection triggers the expression of KC-like and further studies are warranted of KC-like as a possible biomarker for diagnosing sepsis and uterine bacterial infection in dogs.

New insights in Staphylococcus pseudintermedius pathogenicity: antibiotic-resistant biofilm formation by a human wound-associated strain.

BackgroundStaphylococcus pseudintermedius is an opportunistic pathogen recognized as the leading cause of skin, ear, and post-operative bacterial infections in dogs and cats. Zoonotic infections have also recently been reported causing endocarditis, infection of surgical wounds, rhinosinusitis, and catheter-related bacteremia. The aim of the present study is to evaluate, for the first time, the pathogenic potential of S. pseudintermedius isolated from a human infection. To this end, strain DSM 25713, which was recently isolated from a wound of a leukemic patient who underwent a bone marrow transplantation, was investigated for biofilm formation and antibiotic-resistance under conditions relevant for wound infection.ResultsThe effect of pH (5.5, 7.1, and 8.7) and the presence of serum (diluted at 1:2, 1:10, and 1:100) on biofilm formation was assessed through a crystal violet assay. The presence of serum significantly reduced the ability to form biofilm, regardless of the pH value tested. In vitro activity of eight antibiotics against biofilm formation and mature 48 h-old biofilms was comparatively assessed by crystal violet assay and viable cell count, respectively. Antibiotics at sub-inhibitory concentrations reduced biofilm formation in a dose-dependent manner, although cefoxitin was the most active, causing a significant reduction already at 1/8xMIC. Rifampicin showed the highest activity against preformed biofilms (MBEC90: 2xMIC). None of the antibiotics completely eradicated the preformed biofilms, regardless of tested concentrations. Confocal and electron microscopy analyses of mature biofilm revealed a complex “mushroom-like” architecture consisting of microcolonies embedded in a fibrillar extracellular matrix.ConclusionsFor the first time, our results show that human wound-associated S. pseudintermedius is able to form inherently antibiotic-resistant biofilms, suggestive of its pathogenic potential, and consistent with recent reports of zoonotic infections.

Pathogenic Escherichia coli and lipopolysaccharide enhance the expression of IL-8, CXCL5, and CXCL10 in canine endometrial stromal cells

Chemokines play a central role in cellular communication in response to bacterial infection. However, the knowledge of the chemokine responses to bacterial infections in dogs remains limited. Uterine bacterial infection (pyometra) is one of the most common bacterial diseases in dogs and causes sepsis in most of the cases. We have shown previously that dogs with pyometra have higher messenger RNA (mRNA) levels of chemokines in uterus. To assess whether the stromal part of the endometrium expresses chemokines in response to bacterial infection, we cultured endometrial stromal cells isolated from healthy dogs and exposed them to either live pathogenic Escherichia coli, isolated from the uterus of a dog with pyometra, or lipopolysaccharide. Changes in the mRNA expression of ELR+ CXC chemokines, IL-8, CXCL5, CXCL7, and ELR− CXC chemokine, CXCL10, were measured after 24 hours using quantitative real-time polymerase chain reaction. Levels of IL-8, CXCL5, and CXCL10 were upregulated in endometrial stromal cells exposed to E coli and lipopolysaccharide, whereas the level of CXCL7 was decreased or unaffected. In addition, levels of IL-8 and CXCL5, but not CXCL7 or CXCL10, were significantly higher in dogs with pyometra than those in healthy dogs. Our findings show that pathogenic uterine-derived E coli induces a CXC chemokine response both in cultured endometrial stromal cells within 24 hours and in pyometra-affected uteri from dogs. Stromal cells could therefore play an important role in early neutrophil and T cell recruitment to the site of inflammation during gram-negative bacterial infection of the uterus. Further studies are needed to clarify the role of chemokines in host response to bacterial infection in dogs and the possibility of using chemokines as diagnostic parameters for bacterial infection in this species.

Studies on the therapy with quinolones in some bacterial diseases in dogs and cats.

Normal 0 MicrosoftInternetExplorer4 Currently , the most used antibacterial substances include fluoroquinolones ( ciprofloxacin, enrofloxacin , marbofloxacin ), which can be administered both orally and injectable, being active against most aerobic Gram- negative bacteria and moderately active against aerobic Gram - positive bacteria. In this study it was evaluated the effectiveness and frequency of quinolones use in the treatment of various bacterial infections in dogs and cats, respectively urinary infections and dermatitis. The stud y was conducted on 22 dogs and cats with bacterial dermatitis and urinary disorders. Following pathogen susceptibility tests, it was found that the isolated bacterial species were sensitive to quinolones , which were considered the first choice of therapeutic schemes. The results confirmed that the use of quinolones is an alternative to the current antibacterial battle .

Evaluation of therapeutic efficacy of medical shampoo containing terbinafine hydrochloride and chlorhexidine in dogs with dermatophytosis complicated with bacterial infection

This study evaluated the efficacy and safety of medical shampoo containing terbinafine hydrochloride and chlorhexidine gluconate in dogs with dermatophytos is complicated with bacterial infection. Although several studies in dogs and cats with fungal dermatitis have found that oral administration of terbinafine is effective for controlling clinical signs, the topical form of terbinafine has rarely been studied in dogs and cats. Therefore, this study evaluated the efficacy of medical shampoo containing terbinafine hydrochloride and chlorhexidine gluconate in dogs with dermatophytos is complicated with bacterial infection. Eight dogs infected with Microsporum spp. complicated with bacterial infection were enrolled in this study. These dogs were further blindly divided into Group 1 (no treatment, fourdogs) and group 2 (treated with medical shampoo with terbinafine/chlorhexidine, four dogs). Clinical improvement was evaluated using bacterial and fungal cultural evaluation combined with clinical evaluation. This study found that the medical shampoo has sufficient efficacy to treat varying degrees of dermatophytosis complicated with bacterial infection in dogs, although the speed of improvement differed according to the degree and type of infection. Our study also found that combined therapy using antifungal and antibacterial agents can improve clinical signs more effectively and rapidly, suggesting that concurrent bacterial infection plays a significant role in the development of dermatitis.

The use of antibiotics to treat bacterial infections in dogs and cats

Bacterial skin disease in the dog and cat is common and treatment can be frustrating. The process of using history, clinical signs, cytology and bacteriology to decide upon appropriate and effective antibiotic therapy is described. Some alternatives to antibiotics are discussed. Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Bactericidal properties of pradofloxacin against veterinary pathogens

Pradofloxacin is a new veterinary 8-cyano-fluoroquinolone developed for use against bacterial infections in dogs and cats involving both aerobic and anaerobic bacteria. The minimal bactericidal concentrations have been determined against clinical isolates of Staphylococcus pseudintermedius, Staphylococcus aureus, Escherichia coli, Pasteurella multocida, Streptococcus canis, Proteus spp., Fusobacterium spp., Porphyromonas gingivalis and Prevotella species. A subset of these species was selected, and the in vitro rate of kill by pradofloxacin was determined. For 27 of the 30 tested aerobic strains the pradofloxacin MBC was within two doubling dilutions of the MIC. For the remaining strains, the MIC and MBC were within three to four doubling dilutions. Pradofloxacin also demonstrated bactericidal activity against all anaerobic strains, and the MBC was equal to the MIC for four of the strains, within 1 doubling dilution for three strains, within 2 dilutions for a further 3 strains and within 3 dilutions for the remaining five strains. As pradofloxacin concentration was increased, a faster rate of killing was observed; bactericidal effects were seen in all cases at concentrations ≤0.25μg/mL. The bactericidal activity against the anaerobic strains was marked, of particular relevance was the complete absence of regrowth even at 48h at concentrations as low as 0.125μg/mL. In conclusion, pradofloxacin exhibits clear bactericidal activity in terms of MBC and kill kinetics against aerobic and anaerobic clinical isolates from dogs and cats at concentrations that are greatly exceeded within the systemic circulation after administration of the recommended therapeutic doses to the target animals. It is expected that such a rapid rate of kill will play a significant role in clinical efficacy. These data demonstrate the complete and rapid killing of anaerobic bacteria by a veterinary 8-cyano-fluoroquinolone.

Differential expression of Toll-like receptor 4 (TLR4) in healthy and infected canine endometrium

This study provides the first report into immunohistochemical localization of Toll-like receptor (TLR) in the canine reproductive tract. TLR4 was investigated in endometrium during the estrous cycle and in pyometra. Pyometra is the most important pathological condition of the uterus due to bacterial infection in dogs. To protect against invading pathogens, the female reproductive tract has evolved immune mechanisms. TLRs are the cellular components of the afferent arm of the innate immune system. The expression of TLR4 was significantly higher in the endometrial stroma compared to the endometrial surface epithelium and glandular epithelium in proestrus. The glandular epithelium and stroma at the diestrous stage expressed TLR4 significantly higher than surface epithelium. Furthermore, when compared to other healthy groups, the glandular epithelium at diestrus also higher expressed TLR4 than other stages. The expression of TLR4 in the surface epithelium was higher in dogs with pyometra compared with all other groups. And, the surface epithelium of dogs suffering from pyometra also expressed TLR4 more intensely than the glandular epithelium. The innate immunity of infected canine endometrium response to bacterial infection is intensely extremely increased by the expression of TLR4. Furthermore, the different levels of TLR4 expression seems related to physiological changes in distinct cell types of endometrium, leukocytes populations, cytokines and sex hormones.