Identification of bacteria associated with feline chronic gingivostomatitis using culture-dependent and culture-independent methods

Feline chronic gingivostomatitis (FCGS) is a chronic inflammatory disease of the oral cavity that causes severe pain and distress. There are currently no specific treatment methods available and little is known regarding its aetiology, although bacteria are thought to play a major role. The purpose of this study was to identify the oral bacterial flora in normal and diseased cats. Oral swabs were obtained from the palatoglossal folds of eight cats (three normal and five FCGS) and were subjected to microbiological culture. Pasteurella pneumotropica and Pasteurella multocida subsp. multocida were the most prevalent species identified by culture methods in the normal and FCGS samples, respectively. Bacteria were also identified using culture-independent methods (bacterial 16S rRNA gene sequencing). For the normal samples, 158 clones were analysed and 85 clones were sequenced. Capnocytophaga canimorsus (10.8% of clones analysed) was the predominant species. Uncultured species accounted for 8.2% of clones analysed, and 43.7% of clones analysed represented potentially novel species. For the FCGS samples, 253 clones were analysed and 91 clones were sequenced. The predominant species was P. multocida subsp. multocida (51.8% of clones analysed). Uncultured species accounted for 8.7% of clones analysed, and 4.7% of clones analysed represented potentially novel species. It is concluded that the oral flora in cats with FCGS appears to be less diverse than that found in normal cats. However, P. multocida subsp. multocida is found to be significantly more prevalent in FCGS than in normal cats and consequently may be of aetiological significance in this disease.     

Correlation between metabolomic profile constituents and feline pancreatic lipase immunoreactivity

BackgroundFeline pancreatic lipase immunoreactivity (fPLI) is commonly used to diagnose pancreatitis in cats (FP). Untargeted metabolomics has been extensively applied in human and veterinary medicine, but no metabolomic studies regarding FP have been conducted.ObjectivesTo identify metabolites significantly associated with increased fPLI.AnimalsForty‐nine client‐owned cats: 11 clinically healthy and 38 with various clinical conditions.MethodsAnalytical cross‐sectional study with convenience sampling. A panel of 630 metabolites belonging to 26 biochemical classes was quantified in plasma using a commercial metabolomic assay. The correlation between plasma metabolite concentrations and serum fPLI was evaluated using Spearman''s rank correlation coefficient (R _s) with Bonferroni correction. Multivariable analysis then was performed to control for glomerular filtration rate, liver damage, and blood glucose concentration. The accuracy of selected metabolites in discriminating between cats with normal (≤3.5 μg/L) and increased (>5.3 μg/L) fPLI was estimated using the area under the receiver operating characteristic curve (AUROC).ResultsFour hundred and seven of 630 metabolites (64.6%) were quantified in all cats. When controlled for potential confounders only 3 sphingolipids were significantly positively correlated with fPLI: 2 cerebrosides: HexCer(d18:1/24:0); (R _s = .56), and HexCer(d18:1/24:1); (R _s = .58) and 1 sphingomyelin: SM C18:0 (R _s = .55). Their AUROCs in identifying cats with increased fPLI were 82% (95% confidence interval [CI 95%], 70%‐94%), 84% (CI 95%, 72%‐96%), and 78% (CI 95%, 65%‐92%), respectively.Conclusions and Clinical ImportanceSelected sphingolipids are moderately positively correlated with fPLI and appear to have fair to moderate diagnostic accuracy in discriminating between cats with normal and increased fPLI.     

The influence of oral bacteria on tissue levels of Toll-like receptor and cytokine mRNAs in feline chronic gingivostomatitis and oral health

Feline chronic gingivostomatitis (FCGS) is an inflammatory disease of the oral cavity that causes severe pain and distress in affected cats. Treatment methods are currently very limited. The aims of this study were to assess the feline innate immune response by investigating the levels of cytokine and Toll-like receptor (TLR) mRNAs in tissue biopsies of cats with and without FCGS, and to relate this to the presence or absence of putative oral pathogens identified previously within these cats. Mucosal biopsies were collected from 28 cats with FCGS and eight healthy cats. The levels of TLR (TLR2, TLR3, TLR4, TLR7, TLR9) and cytokine (IL-1β, IL-4, IL-6, IL-10, IL-12, TNF-α, IFN-γ) mRNA was determined using quantitative PCR. In the FCGS group a statistically significant increase was seen in TLR2, TLR7, TNF-α, IFN-γ, IL-1β and IL-6 mRNA levels compared to the healthy group. In cats where Tannerella forsythia was present, statistically significant increases were seen in TLR2, TLR4, TLR7, TLR9, TNF-α and IL-1β mRNA levels compared to cats where this putative pathogen was absent. Statistically significant increases in mRNA expression were also seen in cats harbouring feline calicivirus (FCV) (TLR2, IL-1β, IL-6, IFN-γ) and Porphyromonas circumdentaria (TLR2, TLR3) compared to cats where these putative pathogens were absent. Pasteurella multocida subsp. multocida and Pseudomonas sp. did not significantly alter the expression of any TLR or cytokine mRNAs when compared to animals who tested negative for these species, while cats colonised with P. multocida subsp. septica demonstrated a statistically significant reduction in the expression of TLR7, TNF-α and IFN-γ mRNAs compared to cats free of this species. The expression of mRNA for several TLRs and cytokines is elevated in FCGS. A positive correlation was observed between clinical disease severity and the presence of FCV (p = 0.001; Rho = 0.58). Although the number of cats harbouring T. forsythia was low by comparison, 80% of samples in which it was present were from cases with the highest clinical disease severity. Positive correlations with clinical disease severity were seen for TLR2 (p = 0.00086), TLR7 (p = 0.049), TNF-α (p = 0.027), IFN-γ (p = 0.0015), IL-1β (p = 0.004) and IL-6 (p = 0.00001) mRNAs. The putative pathogens FCV and T. forsythia may be important in stimulating a host immune response to FCGS and may play a role in the pathogenesis of this disease.     

The Effect of Experimental Methyl Mercury Poisoning on the Distribution of Acid Phosphatase During Autolysis in Cat Liver

Seven healthy male cats weighing between 2.35 and 4.30 kg were daily fed a diet which contained Hg²⁰³ labelled methyl mercury hydroxide in liver homogenate. Eight additional cats were kept as controls on a similar diet without methyl mercury hydroxide. The daily amount of methyl mercury hydroxide fed to the cats, expressed as inorganic mercury, varied between 3.75 and 4.33 mg per cat. When the animals had developed neurological symptoms typical of methyl mercury poisoning, they were decapitated and their livers removed for the determination of the mercury content, the distribution of acid phosphatase during autolysis at 37°G, the pH and the total bacterial count before and after a 24 hr. period of autolysis. Similar determinations except for the mercury were made from the livers of the control animals. The total amount of methyl mercury hydroxide fed to the cats varied between 29.14 and 40.12 mg Hg++ per kg of body weight, and the mercury content in their livers between 102.5 and 128.7 mg Hg++ per kg of liver wet weight.The share of un sedimentable activity of acid phosphatase out of the total immediately after decapitation was found to be significantly (P < 0.001) greater in the livers of the methyl mercury-fed cats than in the livers of the control animals. After 12 to 24 hrs. of autolysis at 37 °G the unsedimentable activity accounted for 100 % of the total acid phosphatase activity in the livers of 6 of the 7 methyl mercury-fed animals, while in the livers of the controls the corresponding percentages varied after 24 hrs. of incubation between 42 and 73, the mean being 57.4 ± 11.4 %. The mean pH of the livers of the methyl mercury fed animals was found to be significantly higher before (P< 0.001) and after (P < 0.001) a 24 hr. incubation at 37 °G than the corresponding mean pH values of the control animals.Because of the injection of antibiotics given to the cats before sacrifice the total bacterial count of the livers, which was checked before and after a 24 hr. incubation at 37°G, was found to be zero.     

A water-soluble β-glucan improves growth performance by altering gut microbiome and health in weaned pigs

Beta-glucan has been shown to have a beneficial effect on gastrointestinal health. This experiment was conducted to investigate the effects of β-glucan isolated from Agrobacterium sp. ZX09 on growth performance and intestinal health of weaning pigs. A total of 108 weaned pigs (21 d of age; 6.05 ± 0.36 kg) were randomly divided into 3 groups (6 pens/group; 6 pigs/pen), and the groups were each treated with the following diets: 1) basal diet, 2) basal diet supplemented with 20 mg/kg olaquindox, 3) basal diet supplemented with 200 mg/kg β-glucan, for 21 d. Compared with the control group, pigs fed with 200 mg/kg β-glucan had greaterBW, average daily gain and duodenal villus height to crypt depth ratio (P < 0.05). Olaquindox increased the duodenal or jejunal villus height of pigs compared with β-glucan. Compared with the control group, β-glucan tended to increase the occludin mRNA expression in the jejunum (0.05 < P < 0.10). Beta-glucan enriched the beneficial microbiota in the ileum of pigs (P < 0.05). In conclusion, β-glucan may promote growth performance by improving intestinal health and increasing beneficial microbiota of weaned pigs. The study results will provide valuable theoretical guidance for the utilization of β-glucan in weaned pigs.     

Treatment of refractory feline sporotrichosis with a combination of intralesional amphotericin B and oral itraconazole

OBJECTIVE To describe the use of intralesional amphotericin B in localised lesions for the treatment of 26 cats from Rio de Janeiro, Brazil, with sporotrichosis refractory to oral itraconazole. DESIGN Uncontrolled intervention study. METHOD The 26 cats in this study were diagnosed with sporotrichosis, confirmed by isolation of Sporothrix schenckii, and presented residual localised skin lesions refractory to treatment with oral itraconazole for a minimum period of 8 weeks. The animals received weekly applications of intralesional amphotericin B in conjunction with oral itraconazole. In cases of owner unavailability, a maximum of 2 weeks between the infiltrations was accepted. RESULTS Twenty-two (84.6%) of the 26 treated cats achieved clinical remission, 16 (72.7%) of which were cured, and in the remaining six (27.3%) the lesions recurred at the same site. Lack of clinical response was observed in one animal and three owners abandoned treatment. CONCLUSION The proposed therapeutic regimen is an adjunctive treatment option for cats with sporotrichosis presenting as residual skin lesions refractory to itraconazole.     

Salmeterol or doxycycline do not inhibit acute bronchospasm and airway inflammation in cats with experimentally-induced asthma

The objective of this study was to determine if inhaled salmeterol, a long-acting β(2)-adrenergic agonist, and oral doxycycline, a tetracycline antibiotic displaying matrix metalloproteinase (MMP) inhibitory activity, reduce airway inflammation and obstruction in cats with experimentally-induced asthma. Eight Ascaris suum (AS)-sensitised cats were enrolled in a prospective study in which they underwent four AS-challenges at 1 month intervals. The challenged animals were given no treatment or were treated on 4 consecutive days with either: (1) oral prednisolone (1mg/kg twice daily), (2) inhaled salmeterol (50 μg twice daily), or (3) oral doxycycline (5mg/kg twice daily), according to a randomised cross-over design. Inhibition of allergen-induced early (EAR) and late (LAR) asthmatic reactions were assessed by barometric whole-body plethysmography. Cytology and measurement of MMP-2 and -9 activities were carried out on bronchoalveolar lavage fluid (BALF). Although none of the treatments prevented the EAR, prednisolone treatment inhibited the LAR. Relative to untreated cats, the eosinophil percentage and MMP-2 activity in BALF were significantly reduced following prednisolone treatment (P<0.05). Short-term therapy with either salmeterol or doxycycline had no effect on the EAR or LAR or on airway inflammation. Given the chronic nature of this disease in cats, long-term therapy may be required to produce more favourable functional and clinical outcomes.     

Prevalence of feline coronavirus types I and II in cats with histopathologically verified feline infectious peritonitis

Feline coronaviruses (FCoV) vary widely in virulence causing a spectrum of clinical manifestations reaching from subclinical course to fatal feline infectious peritonitis (FIP). Independent of virulence variations they are separated into two different types, type I, the original FCoV, and type II, which is closely related to canine coronavirus (CCV). The prevalence of FCoV types in Austrian cat populations without FIP has been surveyed recently indicating that type I infections predominate. The distribution of FCoV types in cats, which had succumbed to FIP, however, was fairly unknown. PCR assays have been developed amplifying parts of the spike protein gene. Type-specific primer pairs were designed, generating PCR products of different sizes. A total of 94 organ pools of cats with histopathologically verified FIP was tested. A clear differentiation was achieved in 74 cats, 86% of them were type I positive, 7% type II positive, and 7% were positive for both types. These findings demonstrate that in FIP cases FCoV type I predominates, too, nonetheless, in 14% of the cases FCoV type II was detected, suggesting its causative involvement in cases of FIP.     

Basic biological characterization of feline morbillivirus

Feline morbillivirus (FmoPV) is an emerging virus that was recently discovered in domestic cats with chronic nephritis. Despite the potential role of FmoPV in chronic nephritis, little is known about its biological characteristics. In this study, we established a quantitative assay of FmoPV by using an indirect immunofluorescence technique. Viral titers of FmoPV were determined in one week. Treatment with polybrene^® or trypsin which was previously used in virus isolation did not augment the virus titers. FmoPV was notably stable at 4°C, retaining high titers for at least 12 days. Heat-treatment at 60°C and 70°C effectively inactivated FmoPV in 10 and 2 min, respectively. The biological characteristics of FmoPV reported here will be beneficial for establishing an efficient virus isolation method and will provide important information to take a measure to reduce the risk of FmoPV infection.     

Spermine protects intestinal barrier integrity through ras-related C3 botulinum toxin substrate 1/phospholipase C-γ1 signaling pathway in piglets

Weaning stress can cause tight junctions damage and intestinal permeability enhancement, which leads to intestinal imbalance and growth retardation, thereby causing damage to piglet growth and development. Spermine can reduce stress. However, the mechanism of spermine modulating the intestinal integrity in pigs remains largely unknown. This study aims to examine whether spermine protects the intestinal barrier integrity of piglets through ras-related C3 botulinum toxin substrate 1 (Rac1)/phospholipase C-γ1 (PLC-γ1) signaling pathway. In vivo, 80 piglets were categorised into 4 control groups and 4 spermine groups (10 piglets per group). The piglets were fed with normal saline or spermine at 0.4 mmol/kg BW for 7 h and 3, 6 and 9 d. In vitro, we investigated whether spermine protects the intestinal barrier after a tumor necrosis factor α (TNF-α) challenge through Rac1/PLC-γ1 signaling pathway. The in vivo study found that spermine supplementation increased tight junction protein mRNA levels and Rac1/PLC-γ1 signaling pathway gene expression in the jejunum of piglets. The serum D-lactate content was significantly decreased after spermine supplementation (P < 0.05). The in vitro study found that 0.1 μmol/L spermine increased the levels of tight junction protein expression, Rac1/PLC-γ1 signaling pathway and transepithelial electrical resistance, and decreased paracellular permeability (P < 0.05). Further experiments demonstrated that spermine supplementation enhanced the levels of tight junction protein expression, Rac1/PLC-γ1 signaling pathway and transepithelial electrical resistance, and decreased paracellular permeability compared with the NSC-23766 and U73122 treatment with spermine after TNF-α challenge (P < 0.05). Collectively, spermine protects intestinal barrier integrity through Rac1/PLC-γ1 signaling pathway in piglets.     

Management of feline diabetes mellitus.

Up to one quarter of diabetic cats can be well controlled with oral hypoglycemic drugs, although at least 75% require insulin therapy. Most available insulins provide good clinical control but only moderate glycemic control. Because mild to moderate hyperglycemia is well tolerated by cats receiving insulin but hypoglycemia can be life threatening, conservative insulin dosing is recommended. Clinical signs and water intake indicate whether a dose adjustment is required, but serial blood glucose measurements are usually needed to determine the direction of the adjustment. Starting doses of 0.3 to 0.5 IU/kg administered twice daily (rounded down to the nearest whole unit) are usually safe. Dose adjustments should not exceed 1 IU per cat every 2 to 4 weeks unless clinical hypoglycemia has occurred. Cats with clinical hypoglycemia need to be reassessed to see if they are in remission. If not, a 50% to 75% reduction in dose is advised. Approximately 30% of cats go into diabetic remission 1 to 4 months after an adequate treatment protocol is instituted.     

Inflammation and wound healing in cats with chronic gingivitis/stomatitis after extraction of all premolars and molars were not affected by feeding of two diets with different omega‐6/omega‐3 polyunsaturated fatty acid ratios

Feline chronic gingivitis/stomatitis (FCGS) is a painful inflammatory disease in cats. Extraction of teeth, including all premolars and molars, has been shown to be the therapy of choice in cats not responding sufficiently to home care (e.g. tooth brushing) and/or medical treatment (corticosteroids and/or antibiotics). In this study, we hypothesize that a cat food with an omega‐6 polyunsaturated fatty acid (ω6 PUFA) to ω3 PUFA ratio of 10:1 reduces inflammation of the FCGS and accelerates soft tissue wound healing of the gingiva after dental extractions, compared to a cat food with a ω6:ω3 PUFA ratio of 40:1. The cats were fed diets with chicken fat and fish oil as sources of fatty acids. In one diet, part of the fish oil was replaced by safflower oil, resulting in two diets with ω6:ω3 PUFA ratios of 10:1 and 40:1. This double‐blinded study in two groups of seven cats revealed that dietary fatty acids influence the composition of plasma cholesteryl esters and plasma levels of inflammatory cytokines. The diet with the 10:1 ratio lowered PGD₂, PGE₂ and LTB₄ plasma levels significantly, compared to the diet with the 40:1 ratio (p = 0.05, p = 0.04, and p = 0.02 respectively). However, feeding diets with dietary ω6:ω3 PUFA ratios of 10:1 and 40:1, given to cats with FCGS for 4 weeks after extraction of all premolars and molars, did not alter the degree of inflammation or wound healing.     

Peripheral and Central Venous Blood Glucose Concentrations in Dogs and Cats with Acute Arterial Thromboembolism

Background

Acute limb paralysis because of arterial thromboembolism (ATE) occurs in cats and less commonly in dogs. ATE is diagnosed based on physical examination findings and, occasionally, advanced imaging.

Hypothesis/Objectives

Peripheral, affected limb venous glucose concentration is decreased in ATE, whereas its systemic concentration is within or above reference interval.

Animals

Client‐owned cats and dogs were divided into 3 respective groups: acute limb paralysis because of ATE (22 cats and 9 dogs); acute limb paralysis secondary to orthopedic or neurologic conditions (nonambulatory controls; 10 cats and 11 dogs); ambulatory animals presented because of various diseases (ambulatory controls; 10 cats and 9 dogs).

Methods

Prospective observational, clinical study. Systemic and local (affected limb) blood glucose concentrations were measured. Their absolute and relative differences (ΔGlu and %ΔGlu, respectively) were compared among groups.

Results

ΔGlu and %ΔGlu were significantly higher in the ATE cats and dogs groups, compared to both of their respective controls (P < .0001 and P < .001, respectively). No significant differences were observed between the control groups. Receiver operator characteristics analysis of ΔGlu and %ΔGlu as predictors of ATE had area under the curve of 0.96 and 0.99 in cats, respectively, and 1.00 and 1.00, in dogs, respectively. ΔGlu cutoffs of 30 mg/dL and 16 mg/dL, in cats and dogs, respectively, corresponded to sensitivity and specificity of 100% and 90% in cats, respectively, and 100% in dogs.

Conclusions and Clinical Importance

ΔGlu and %ΔGlu are accurate, readily available, diagnostic markers of acute ATE in paralyzed cats and dogs.     

Serologic and Urinary PCR Survey of Leptospirosis in Healthy Cats and in Cats with Kidney Disease

Background

Although there is serologic evidence of exposure of cats to Leptospira spp., clinical disease is rarely reported in cats.

Objective

To compare the seropositivity and urinary polymerase chain reaction (PCR) status for Leptospira spp. between healthy (H) cats and cats with kidney disease (KD), to investigate the serovars potentially involved, and to evaluate potential risk factors.

Animals

Two hundred and forty client‐owned cats.

Methods

Cats were prospectively recruited and classified based on physical examination, complete blood count, serum biochemistry profile, and urinalysis (125 H and 115 KD cats). Leptospira spp. serology (titers ≥1 : 100 considered positive) and urinary PCR were performed in all cats. Data assessing risk factors, obtained from a questionnaire, were evaluated using logistic regression models.

Results

Seropositivity for Leptospira spp. was statistically different between groups: 7.2% (9/125) and 14.9% (17/114) in the H and KD, respectively (P = .05). The proportion of PCR‐positive cats was not. The most common serovars detected serologically were Pomona (n = 16) and Bratislava (n = 8). Risk factors for seropositivity included outdoor and hunting lifestyles (P = .03 and P < .001, respectively), the presence of another cat in the household (P < .01), and the sampling period, with the greatest number of cases identified between June and August (P =.02).

Conclusions

Seropositivity was significantly greater in KD cats, suggesting that the role of Leptospira spp. in KD in cats should be further investigated. The detection of urinary shedding of leptospires in several cats identifies a potential role in the transmission of the organism.     

Pulmonary lesions in cats with diabetes mellitus

Diabetes mellitus (DM) is a common endocrinopathy of cats and humans. Although few studies have examined the effects of DM on the pulmonary system, changes in pulmonary function and immunology in humans with type I and II diabetes, and pulmonary lesions in a murine diabetic model have been documented. Our objective was to determine whether pulmonary lesions occurred in cats with DM. Medical records and necropsy evaluations of 42 cats with DM were compared with those of 45 age-matched, nondiabetic cats for the presence of clinical evidence of respiratory disease and pulmonary histopathological findings at the time of necropsy. No statistical difference was noted in the presence of clinical evidence of respiratory disease between cats with diabetes and control cats. Nevertheless, there was a significant association between the presence of abnormal pulmonary histopathology and DM (P = .018, odds ratio = 3 inclusive of all cats; P = .005, odds ratio = 5 when non-DM cats with overt clinical evidence of respiratory disease were excluded). Pulmonary abnormalities detected by histopathological examination in cats with diabetes included congestion and edema, histiocytosis, pneumonia, smooth muscle hypertrophy, fibrosis, mineralization, neoplasia, and type II pneumocyte hyperplasia. The observed association between DM and pulmonary lesions in cats, independent of clinical evidence of respiratory disease, emphasizes the need for careful assessment of the respiratory tract in sick cats with diabetes.     

Total sialic acid: An acute phase reactant in cats with a possible role in feline coronavirus infection

The aims of this study were to validate a colorimetric method to measure total sialic acid (TSA) in feline serum and to investigate the serum concentration of TSA in clinically healthy cats seronegative (n = 9) and seropositive (n = 48) for feline coronavirus (FCoV), and in cats affected by feline infectious peritonitis (FIP, n = 28), tumors (n = 20), or inflammation (n = 16). The correlation between TSA and α₁-acid glycoprotein (AGP) was also investigated. The method employed in this study is precise and accurate at TSA levels (in mg/L) commonly encountered in feline serum. No significant differences between seropositive (385.6 ± 192.2 mg/L) and seronegative (433.5 ± 179.0 mg/L) cats were detectable, suggesting that the simple infection by FCoVs does not influence TSA levels. Compared with seropositive controls, the concentration of TSA was higher in cats with FIP (556.7 ± 268.3 mg/L, P = 0.003), tumors (522.5 ± 294.4 mg/L, P = 0.028), and inflammation (546.8 ± 208.3 mg/L, P = 0.018). The discriminating power of TSA for FIP is moderate (area under the ROC curve = 0.65) and the likelihood ratio is higher than 3.0 only at high TSA levels. Consequently, TSA could support a diagnosis of FIP only at extremely high serum concentration (> 800 mg/L) or when the pre-test probability of FIP is high. No correlations were found between the TSA and AGP concentrations in cats with FIP, suggesting that sialylated proteins other than AGP are present. Both the antibody titre and the degree of AGP sialylation were negatively correlated with TSA levels, suggesting that increased TSA may contribute to reduce the burden of FCoVs.