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24-hour evaluation of dental plaque bacteria and halitosis after consumption of a single placebo or dental treat by dogs
Abstract
OBJECTIVE To determine whether consumption of a single dental treat with specific mechanical properties and active ingredients would provide a 24-hour effect on dental plaque bacteria and halitosis in dogs.
ANIMALS 10 dogs of various breeds from a privately owned colony that had received routine dental scaling and polishing 4 weeks before the study began.
PROCEDURES Dogs were randomly assigned to receive 1 placebo or dental treat first. A 4-week washout period was provided, and then dogs received the opposite treatment. Oral plaque and breath samples were collected before and 0.5, 3, 12, and 24 hours after treat consumption. Volatile sulfur compounds (VSCs) concentration was measured in breath samples. Total aerobic, total anaerobic, Porphyromonas gulae, Prevotella intermedia–like, Tannerella forsythia , and Fusobacterium nucleatum bacterial counts (measured via bacterial culture) and total live bacterial counts, total live and dead bacterial counts, and bacterial vitality (measured via quantitative real-time PCR assay) were assessed in plaque samples.
RESULTS Compared with placebo treat consumption, dental treat consumption resulted in a significant decrease in breath VSCs concentration and all plaque bacterial counts, without an effect on bacterial vitality. Effects of the dental treat versus the placebo treat persisted for 12 hours for several bacterial counts and for 24 hours for breath VSCs concentration.
CONCLUSIONS AND CLINICAL RELEVANCE Although clinical benefits should be investigated in larger scale, longer-term studies, results of this study suggested that feeding the evaluated dental treat may help to decrease oral bacterial growth in dogs for 12 hours and oral malodor for 24 hours. A feeding interval of 12 hours is therefore recommended.
... Several studies (Quirynen et al., 2006;Jeusette et al., 2016;Oba et al., 2021) investigated the effectiveness of dog treats at promoting canine oral health. One study (Jeusette et al., 2016) found that a specific dental treat containing vitamin C and zinc sulfate reduced oral bacterial growth and malodor in dogs for up to 12 and 24 hours, respectively. ...
... Several studies (Quirynen et al., 2006;Jeusette et al., 2016;Oba et al., 2021) investigated the effectiveness of dog treats at promoting canine oral health. One study (Jeusette et al., 2016) found that a specific dental treat containing vitamin C and zinc sulfate reduced oral bacterial growth and malodor in dogs for up to 12 and 24 hours, respectively. This is significant as PD, a common and widespread oral disease in dogs, has been associated with other health issues (Poppi et al., 2023). ...
The global growth in canine population has led to a thriving market for pet food, especially treats sector. This review explores the growing market for canine treats, revealing a deep emotional connection between pet owners and their pets, evident in feeding practices. With six distinct categories of treats, meat-based treats dominate due to perceptions of naturalness and health, high-lighting an increased focus on dog nutrition and a preference for raw, locally sourced alternatives. However, labeling discrepancies and a lack of clear nutritional information present a challenge for owners looking for quality treats. Variations in nutritional values and sensory qualities of treats require informed choices to prevent excessive calorie consumption and potential health implications. Effective communication between the veterinarian and pet owners is crucial to adapting treats recommendations based on the individual needs of dogs, taking into account potential health risks and obesity. In conclusion, the use of treats provides several benefits such as positive reinforcement, promoting dental health and providing nutritional support for senior dogs. However, certain aspects that may limit their use should be considered, such as the risk of microbiological contamination, accidents related to incorrect administration, obesity and environmental impacts associated with treat production.
... Despite the importance of regular oral care procedures, many dog owners fail to brush their dog's teeth or arrange for routine cleaning by a veterinary professional (Miller and Harvey, 1994;Enlund et al., 2020). However, because most owners give treats to their pets (Morelli et al., 2020), dental chews may serve as an alternative and play a significant role in promoting periodontal health and reducing oral malodor Brown and McGenity, 2005;Clarke et al., 2011;Quest, 2013;Jeusette et al., 2016;Carroll et al., 2020;Ruparell et al., 2020;Oba et al., 2021). Studies have shown that incorporating dental chews into a dog's feeding regime results in significant reduction in plaque and calculus accumulation, as well as gingivitis, and promotes a positive shift in the oral microbiota (Carroll et al., 2020;Ruparell et al., 2020;Oba et al., 2021). ...
... Furthermore, all interventions led to a significant reduction in both the total bacterial load and VSC-producing bacterial load in plaque when compared with the control group (Croft et al., 2022). Those data agreed with previous studies showing that dental chew consumption slows the buildup of plaque as well as oral VSC concentrations (Quest, 2013;Jeusette et al., 2016;Mateo et al., 2020). ...
Halitosis in dogs is an initial indication of periodontitis, highlighting its significance as a vital marker for underlying problems. Moreover, the oral microbial population has a significant influence on periodontal disease. Measuring the oral microbiota may be used in addition to breath odor, dental plaque, and gingivitis scoring to assess the impact of dental chews on oral health. In this study, we aimed to determine the differences in breath odor, oral health outcomes, and oral microbiota of adult dogs consuming a novel dental chew compared with control dogs consuming only a diet. Twelve healthy adult female beagle dogs were used in a crossover design study. Treatments (n = 12/group) included: diet only (control) or the diet + a novel dental chew. Each day, one chew was provided 4 h after mealtime. On days 1, 7, 14, 21, and 27, breath samples were analyzed for total volatile sulfur compound concentrations using a halimeter. On day 0 of each period, teeth were cleaned by a veterinary dentist blinded to treatments. Teeth were scored for plaque, calculus, and gingivitis by the same veterinary dentist on day 28 of each period. After scoring, subgingival and supragingival plaque samples were collected for microbiota analysis using Illumina MiSeq. All data were analyzed using SAS (version 9.4) using the Mixed Models procedure, with P < 0.05 being significant. Overall, the dental chews were well accepted. Dogs consuming the dental chews had lower calculus coverage, thickness, and scores, lower gingivitis scores, and less pocket bleeding than control dogs. Breath volatile sulfur compounds were lower in dogs consuming the dental chews. Bacterial alpha-diversity analysis demonstrated that control dogs had higher bacterial richness than dogs fed dental chews. Bacterial beta-diversity analysis demonstrated that samples clustered based on treatment. In subgingival and supragingival plaque, control dogs had higher relative abundances of potentially pathogenic bacteria (Pelistega, Desulfovibrio, Desulfomicrobium, Fretibacterium, Helcococcus, and Treponema) and lower relative abundances of genera associated with oral health (Neisseria, Actinomyces, and Corynebacterium). Our results suggest that the dental chew tested in this study may aid in reducing periodontal disease risk in dogs by beneficially shifting the microbiota population and inhabiting plaque buildup.
... Studies have demonstrated that within minutes after polishing, approximately one million organisms are deposited per square millimeter of enamel surface. 1 In this context, prophylaxis performed by the pet owner of the animal in the home environment (homecare), in association with specialized care in the clinic, constitutes an effective procedure for the control of PD. 1 The use of physical methods (such as regular tooth brushing, and provision of abrasive foods and toys) together with chemical methods (such as oral antiseptic-containing dentifrices) should be considered as a critical measure for microbial burden reduction and to control proliferation and establishment of these microbial colonies. [5][6][7][8] Solutions, creams, pastes, gels, and foams are the most commonly encountered pharmaceutical dosage forms for use in toothpastes or mouthwashes for the oral cavity. 1,7,8 However, the exposure time of the oral mucosa to the drugs in these products is short, whether due to the washing and rinsing process, or the presence of a continuous salivary flow and the mechanical movements of the mouth and tongue. ...
... [5][6][7][8] Solutions, creams, pastes, gels, and foams are the most commonly encountered pharmaceutical dosage forms for use in toothpastes or mouthwashes for the oral cavity. 1,7,8 However, the exposure time of the oral mucosa to the drugs in these products is short, whether due to the washing and rinsing process, or the presence of a continuous salivary flow and the mechanical movements of the mouth and tongue. 9 Therefore, ointments with mucoadhesive properties, called orabases, are more suitable for local release of drugs into the mucosa. ...
Herbal therapies are used worldwide to treat a variety of health conditions, including dental conditions in veterinary medicine. In this context, the use of medicinal plant-based formulations as potential therapeutics and preventatives in veterinary dentistry is worth highlighting. The objective of the present study was to develop a mucoadhesive ointment formulation, named orabase, that contained pomegranate extract for use in the oral cavity of dogs, with the aim of improving their oral hygiene. The hydroalcoholic extracts of pomegranate peels was incorporated into the orabase in 3 different concentrations. The formulations were subjected to in vitro microbiological testing by a modified disc-diffusion method to study the susceptibility of microorganisms collected from the oral cavities of the dogs. The samples were taken from the buccal mucosa of dogs having the same management and diet. The most effective formulation was submitted to physicochemical tests to evaluate the functionality of the product, namely pH, swelling index, spreadability, and mechanical properties (hardness, cohesiveness, and adhesiveness). The formulation containing 25.0% w/w of the extract was considered most suitable for the intended use as it showed antiseptic activity and demonstrated a swelling index of approximately 35% in the first 20 minutes of the test, high spreadability, and suitable mechanical properties. The results suggest that the product obtained from pomegranate peel extract is a viable option for use to improve oral hygiene, helping to reduce the bacterial component of dental plaque in dogs.
... Bacteria of the genera Actinomyces, Porphyromonas, Fusobacterium, Neisseria, Staphylococcus, Streptococcus and Veillonella were isolated in most samples analyzed by Dent and Marsh [19], Kim et al. [22], Zambori et al. [23] and Jeussete et al. [24], suggesting that representatives of these genera can be considered as characteristic or indigenous members of the microbial community in dental plaque. This was confirmed in the present study, which verified the presence of the genera Staphylococcus, Streptococcus, Actinomyces and Neisseria (as shown in Figure 1). ...
... This was confirmed in the present study, which verified the presence of the genera Staphylococcus, Streptococcus, Actinomyces and Neisseria (as shown in Figure 1). As expected, the genera Porphyromonas, Fusobacterium and Veillonella, found in other works [19,22,23,24] were not isolated as result of the methodology applied and the nutritional and strict anaerobic needs of these bacteria. ...
... The articles evaluating chlorhexidine (16.67%) evaluated the bacteriostatic action in the prevention and treatment of diseases related to plaque formation, and the different routes of application: liquid solution, in the form of mouthwashes and gels, sprays, and mucoadhesive or dental chews. The lowest chlorhexidine level evaluated was 0.12% (Jeusette et al., 2016) and the highest was 20% (Kozlovsky et al., 1992). The absorption of chlorhexidine is minimal, and it has low toxicity. ...
The expected contribution of this study is to have a complete view of the available products used in the prevention of periodontal disease in dogs. The related studies in databases including Web of Science, Science Direct, and Scopus, from 1992 to 2021, were retrieved, and 36 studies were used in the systematic review. Among the continents researched, North America proved to have the most developed research relating to the subject. The active ingredients researched, isolated, or compared to each other, were polyphosphates > phytogenics > chlorhexidine > enzymes > vitamin C > xylitol = adelmidrol = and probiotics. The form of use of active products was chewable > solution via water = via food > biscuits > paste and gels > mucoadhesive. Gingivitis index was the most studied parameter used to assess oral health. According to the evidence recovered in the systematic review was possible to conclude that products with different active agents and forms of use demonstrated positive effects in the prevention of periodontal diseases. The results of this study summarize the most frequently used active ingredients and application forms for cleaning dogs' teeth and provide a roadmap for future research on the maintenance of oral health in dogs.
... Currently, chewing materials are mainly recommended to improve oral health, as food texture is considered to have an influence (Logan, 2006). A range of chewing materials have been shown to be beneficial: several raw hide products (Lage et al., 1990;Stookey, 2009), chew sticks (Gorrel et al., 1999;Hennet et al., 2006;Quest, 2013;Jeusette et al., 2016) and raw beef bones (Marx et al., 2016) reduced plaque and calculus. Therefore the AAHA dental care guidelines recommend dental chews for home dental care (Holmstrom et al., 2013). ...
Chewing is a behavioural element of feeding, but dogs also chew on or dissect non-edible items. This can cause considerable problems to owners if directed at household objects. Nevertheless, the provision of chewing material, associated risks and relationships with chewing behaviour and other owner-dog activities have not been investigated so far. The aim of this online survey was to explore how dog owners manage the chewing behaviour of their dog and whether there are relationships to other owner-dog interactions. Of our self-selected participants (1439 filled in the entire questionnaire), 94% provided their dogs with edible chewing material (e.g., rawhide, dried innards, meat), 83% provided inedible chew toys, 73% provided chew toys filled with food and 51% provided hard chewing material (e.g. wood, antlers). Edible materials were provided four to six times a week by the average dog owner. Regarding risks, 67% of respondents stated that their dog never had a problem caused by the use of chewing material, whereas veterinary treatment due to a problem with chewing material was reported by 3.6%. Chewing daily on soft household objects was observed in 2.5% of dogs (other common objects for daily chewing: resting places 2.2 %, clothes/shoes 1.4%); dogs up to one year of age did this more frequently (p < 0.001). Chewing on objects was not substantially related to reported motivation of the dog to play or the frequency of activities with the dog (all rs < 0.2), but was reported to occur in contexts that may cause negative emotional states such as leaving the dog alone (rs = 0.63, p < 0.001) or changes in routine activities (rs = 0.47, p < 0.001). The average reported frequency of provision of chewing material correlated positively (rs ≥ 0.2) with motivation of the dog to play, chewing on objects, human-dog play and calm activities such as petting. Dog owners think that chewing material is important for puppies and even more for adult dogs (p < 0.001). However, it remains to be investigated how motivated dogs are for chewing on different types of materials and whether chewing, as proposed by dog professionals, reduces stress. This seems particularly important for assessing the trade-off between risks and benefits of different chewing materials and its impact on dog welfare.
... 32,34,35 In a previous study when dogs were given a similar dental chew as used in this study, also containing vitamin C and zinc salts but designed with a different shape, the results showed an antimicrobial effect persisting for 12 to 24 hours after administration of one chew. 36 Sodium phosphate salts (ie, sodium hexametaphosphate) are calcium chelators shown to prevent calculus formation in dogs. 37 They act by inhibiting calcium hydroxyapatite crystal growth and dental plaque mineralization. ...
The aim of this study was to determine the clinical efficacy of a dental chew with mechanical and chemical properties in 2 toy dog breeds. Eight Yorkshire terriers and 9 Chihuahuas participated in a crossover design trial. Gingivitis, dental plaque, calculus, and volatile sulfur compounds (VSC) in the breath were assessed after 4 weeks and 9 weeks, respectively. When dogs were fed 1 dental chew per day they had significantly reduced gingivitis (−20%, P < .001), accumulation of plaque (−15%, P < .001), calculus (−35%, P = .001), and VSC concentration (−19%, P < .001) compared to when receiving no chew. A significant breed effect was observed on all the average dental indices and VSC concentration.
... Feeding a dental chew six days per week has been shown to maintain low breath sulfur for up to 21 months (15). A once-daily chew regimen appears to control oral malodor as it preserved a low, flat diurnal pattern of breath sulfur (18). All the chews that improved halitosis also reduced accumulation of dental deposits, just like severity of gingivitis (6,(15)(16)(17). ...
Halitosis represents a cause of social problems despite of the availability of commercial aids to counteract it. A dietary approach represents an innovative solution. The aim of this clinical evaluation was to assess the efficacy of a commercially available nutraceutical diet in a cohort of dogs suffering from chronic halitosis and manifesting at least one symptom among drooling, gingivitis, ulcers, stomatitis, laryngitis, tartar, dental caries, abscess, lingual lesions. Dogs were fed the nutraceutical diet over a period of 30 days, receiving a veterinary inspection before and at the end of the evaluation. Most of symptoms resulted in significant improvement at the end of the evaluation period. We hypothesize that halitosis may be a consequence of a daily intake of food contaminated with oxytetracycline.
Background:
The pathogenesis and treatment of cutaneous malodour in dogs have not been investigated previously. Staphylococcus and Corynebacterium spp. are associated with human axillary malodour.
Hypothesis:
Staphylococcus and Corynebacterium spp. are associated with cutaneous malodour in dogs, and treatment with a topical essential oil-based product will improve malodour and reduce the abundance of odour-causing bacteria.
Animals:
Twenty seven bloodhound dogs from a south Texas boarding facility were enrolled in this study.
Methods and materials:
Skin swabs were taken from the axilla and dorsum of 27 dogs at initiation of the study. Mean malodour scores were used to assign dogs to control or malodour groups. The malodourous dogs were randomly assigned to a treatment or placebo group, received four weekly topical applications of the spot-on or placebo, and samples were recollected. Next-generation sequencing (NGS) and real-time quantitative PCR (qPCR) were performed on all swabs.
Results:
Psychrobacter and Pseudomonas spp. were significantly more abundant (P < 0.001, P = 0.006; respectively), and overall bacterial diversity was reduced (P = 0.0384) on the skin of malodourous dogs. Staphylococcus and Corynebacterium spp. were not associated with malodour. The topical essential oil-based product significantly (P = 0.0078) improved malodour in the treatment group and shifted their bacterial community structure.
Conclusions and clinical importance:
A novel association of bacterial genera with malodour in bloodhound dogs, identified by NGS, highlights future targets for odour control. The topical treatment significantly reduced malodour. The interaction between the topical treatment and cutaneous microbiota should be further investigated and may be useful in other dermatological conditions involving microbiota.
Bovine foot rot (BFR) is an infectious disease of the interdigital skin and subcutaneous tissues of beef and dairy cattle that occurs under a variety of management and environmental settings. The anaerobic, gram-negative bacteria Fusobacterium necrophorum, Porphyromonas levii, and Prevotella intermedia are commonly isolated from lesions. A multitude of host, agent, and environmental factors contribute to the development of BFR. Initiation of systemic antimicrobial therapy early in the course of disease commonly leads to resolution. Delays in treatment may result in extension of infection into deeper bone, synovial structures, or ligamentous structures, and the prognosis for recovery is reduced.
Background
It has been shown that the prevalence of both clinical attachment loss (CAL) >1 mm and pocket probing depth (PPD) >4 mm is relatively high even in younger dogs, but also that only a minority of the dogs have such clinical signs of periodontal disease (PD) in more than a few teeth. Hence, a minority of dogs carry the major PD burden. These epidemiological features suggest that screening for PD in larger groups of dogs, allowing for rapid assessment of treatment planning, or for the selection of dogs with or without PD prior to be included in experimental trials, should be possible. CAL is the central variable in assessing PD extent and severity while PPD is the central variable used in treatment planning which make these two variables obvious in a screening protocol with the dual aim of disease identification and treatment planning. The main purpose of the present study in 98 laboratory Beagle dogs was to construct a fast, simple and accurate screening tool, which is highly sensitive for the identification of dogs with PD.ResultsExamination of the maxillary P4, P3, P2, I1 and C would, in this population, result in the identification of 85.5% of all dogs and 96% of all teeth positive for CAL ¿1mm, and 58.9% of all dogs and 82.1% of all teeth positive for PD ¿4 mm.Examination of tooth pairs, all C¿s, maxillary I2, M2 and the mandibular P4 would, in this population result in identification of 92.9% of all dogs and 97.3% of all teeth positive for PD ¿4 mm, and 65.5% of all dogs and 83.2% of all teeth positive for CAL ¿1mm. The results presented here only pertain to the present study population.Conclusions
This screening protocol is suitable for examination of larger groups of laboratory Beagle dogs for PD and our findings indicate that diseased dogs are identified with a high degree of sensitivity. Before this screening can be used in clinical practice, it has to be validated in breeds other than Beagle dogs and in populations with larger age variation.
Purebred dog health is thought to be compromised by an increasing occurence of inherited diseases but inadequate prevalence data on common disorders have hampered efforts to prioritise health reforms. Analysis of primary veterinary practice clinical data has been proposed for reliable estimation of disorder prevalence in dogs. Electronic patient record (EPR) data were collected on 148,741 dogs attending 93 clinics across central and south-eastern England. Analysis in detail of a random sample of EPRs relating to 3,884 dogs from 89 clinics identified the most frequently recorded disorders as otitis externa (prevalence 10.2%, 95% CI: 9.1-11.3), periodontal disease (9.3%, 95% CI: 8.3-10.3) and anal sac impaction (7.1%, 95% CI: 6.1-8.1). Using syndromic classification, the most prevalent body location affected was the head-and-neck (32.8%, 95% CI: 30.7-34.9), the most prevalent organ system affected was the integument (36.3%, 95% CI: 33.9-38.6) and the most prevalent pathophysiologic process diagnosed was inflammation (32.1%, 95% CI: 29.8-34.3). Among the twenty most-frequently recorded disorders, purebred dogs had a significantly higher prevalence compared with crossbreds for three: otitis externa (P = 0.001), obesity (P = 0.006) and skin mass lesion (P = 0.033), and popular breeds differed significantly from each other in their prevalence for five: periodontal disease (P = 0.002), overgrown nails (P = 0.004), degenerative joint disease (P = 0.005), obesity (P = 0.001) and lipoma (P = 0.003). These results fill a crucial data gap in disorder prevalence information and assist with disorder prioritisation. The results suggest that, for maximal impact, breeding reforms should target commonly-diagnosed complex disorders that are amenable to genetic improvement and should place special focus on at-risk breeds. Future studies evaluating disorder severity and duration will augment the usefulness of the disorder prevalence information reported herein.
Periodontal disease is the most widespread oral disease in dogs which if left untreated results in significant pain to the pet and loss of dentition. The objective of this study was to identify bacterial species in canine plaque that are significantly associated with health, gingivitis and mild periodontitis (<25% attachment loss). In this survey subgingival plaque samples were collected from 223 dogs with healthy gingiva, gingivitis and mild periodontitis with 72 to 77 samples per health status. DNA was extracted from the plaque samples and subjected to PCR amplification of the V1-V3 region of the 16S rDNA. Pyrosequencing of the PCR amplicons identified a total of 274 operational taxonomic units after bioinformatic and statistical analysis. Porphyromonas was the most abundant genus in all disease stages, particularly in health along with Moraxella and Bergeyella. Peptostreptococcus, Actinomyces, and Peptostreptococcaceae were the most abundant genera in mild periodontitis. Logistic regression analysis identified species from each of these genera that were significantly associated with health, gingivitis or mild periodontitis. Principal component analysis showed distinct community profiles in health and disease. The species identified show some similarities with health and periodontal disease in humans but also major differences. In contrast to human, healthy canine plaque was found to be dominated by Gram negative bacterial species whereas Gram positive anaerobic species predominate in disease. The scale of this study surpasses previously published research and enhances our understanding of the bacterial species present in canine subgingival plaque and their associations with health and early periodontal disease.
Oral disorders of the dog represent for veterinarians a medical challenge and an important field of interest from the economical point of view. Although many epidemiological studies on dental diseases in beagles bred under controlled conditions have been realized, information on frequency of these alterations in populations of pet dogs, especially in Central Europe, is far from complete. The aim of our study was to assess the prevalence of the most common oral diseases in dogs in a Czech urban region. A total number of 408 dogs, presented at a private Czech urban veterinary hospital for different reasons, were analyzed. Site specificity and severity of dental diseases were assessed using modified indexing systems. Dental alterations could be found in 348 out of 408 dogs (85.3%). The most frequent diseases were (i) pe riodontitis (60.0% of 408 dogs), (ii) calculus (61.3%), (iii) missing teeth (33.8%), and (iv) abnormal attrition (5.9%). Furthermore, single cases of caries, tumors and enamel hypoplasia could be observed. Periodontitis occurred preferentially in the upper jaw of small dogs and increased with age. The labial/buccal side of teeth was affected more severely than the lingual/palatinal side. Differences between left and right side could not be observed. Malocclusion and insufficient oral hygiene care seem to predis- pose to periodontitis. As periodontitis, dental calculus occurred preferentially in small dogs and increased with age. The prevalence of calculus formation did not differ between left and right side. However, the upper jaw showed a higher degree of affection than the mandible. On the labial/buccal side of the teeth, a thicker calculus layer could be observed than lingually/palatinally. Interestingly, the degree of calculus formation and of periodontitis did not correlate in all cases, supporting the hypothesis that supragingival calculus per se is not an irritant. The pattern of tooth loss was the same between left and right side and between upper and lower jaw. Most commonly, the first premolars were missing followed by incisors and other premol ars and molars. Tooth loss for other reasons than periodontitis and single cases of tooth agenesis has not been detected in our study. (Abnormal) tooth wear was detected only in older dogs and affected mostly canines and premolars of large breeds. Age estimation based on dental attrition should be carried out with care, because tooth wear depends on keeping conditions and feeding of the dog. Our study confirmed the high prevalence of oral diseases in dogs. Veterinarians could improve the effectiveness of treatment concentrating their diagnostic efforts on age groups and types of teeth at highest risk, as assessed in this and other reports.
We analyzed the distribution of 11 periodontitis-related bacterial species in dental plaque collected from 176 Japanese dogs divided into young (less than 2 years of age), middle-aged (2-7 years of age) and elderly (more than 8 years of age) groups using a polymerase chain reaction method. Clinical examination revealed that no dogs in the young group were affected by periodontitis, whereas the rates for gingivitis and periodontitis were high in the middle-aged and elderly groups. In addition, the total numbers of bacterial species in the middle-aged and elderly groups were significantly greater than in the young group. Our findings suggest that age is an important factor associated with the distribution of periodontitis-related bacteria and periodontal conditions in dogs.
Halitosis (bad breath) is a common condition that is socially crippling for vast parts of the population and results from malodorous volatile sulfur compounds, which are by-products of oral bacteria. In this doubled-blind, randomized study, 75 subjects with halitosis were evaluated. The participants were treated with or without abrasive microcapsules (candy) containing zinc gluconate 0.5%, propolis 2%, and a combination of both (zinc 0.25% and propolis 1%). The halitosis was assessed by a Halimeter, a portable instrument that measures the emission of volatile sulfur compounds at different time exposures to the treatments. Breezy candy (which is sugar-free) was found effective in the treatment of halitosis for up to 4 hours. While treatment with regular candy (group 1, traditional candy without abrasive particles) showed reduction in malodor of 10%, Breezy candy showed reduction of up to 60% in malodor (P < .0001). Since this was not a longitudinal study, the total duration of the effect was not assessed. Breezy candy in the abrasive form with zinc additive had the best potential to positively affect malodor when treating patients with halitosis. The combined effect of abrasion by microcapsules with zinc supplement represents a novel and successful approach for the treatment of halitosis. (Quintessence Int 2011;42:313?317).
A rapid PCR approach was developed to detect Porphyromonas gulae strains from subgingival samples of dogs with and with periodontitis. The presence of P. gulae was observed in 92% and 56%, respectively, in dogs with and without periodontitis. The new primer pair was specific to detect this microorganism, and this technique could be used to evaluate a correlation between periodontitis and P. gulae in companion animals.
To estimate the prevalence and describe the extent and severity of periodontal disease and associated periodontal parameters in beagle dogs.
A full-mouth, site-specific examination was performed in 98 beagle dogs. Focus was placed on clinical attachment loss, pocket depth and bleeding on probing.
The prevalence of clinical attachment loss greater than equal to 1 mm was 20 per cent in the one-year-old dogs, increasing to 84 per cent of the dogs aged more than three years. The number of sites affected with clinical attachment loss greater than equal to 1 mm showed a skewed distribution. The prevalence of clinical attachment loss greater than equal to 4 mm was only seven per cent. A probing pocket depth of 4+ mm was observed in 44 to 81 per cent of the dogs, depending on age. Also, the distribution of the number of deepened pockets/dog was skewed. The teeth most prone to clinical attachment loss greater than equal to 1 mm were the P2, the P3 and the P4 of the maxilla. The teeth most prone to pocket depth greater than equal to 4 mm were the maxillary canines.
Periodontal disease in terms of clinical attachment loss greater than equal to 1 mm and pocket depth greater than equal to 4 mm is common in beagle dogs, but the major disease burden is carried by only a few dogs. The prevalence increases with increased age but is high already at the age of two years.
A study was undertaken to determine the effect of a new dental hygiene chew on periodontal health in the dog. The textural properties of this chew are different from previously tested dental hygiene products. The accumulation of dental deposits, development of oral malodor, and development of gingivitis were assessed in two groups of dogs; one fed a dry diet only, and the other fed the same dry diet supplemented by the daily addition of the new dental hygiene chew. Daily addition of the chew to the dry diet was effective in reducing plaque and calculus accumulation on the tooth surfaces, and also reduced the severity of gingivitis and oral malodor as compared to feeding the dry diet only.
Zinc ions, chlorhexidine (CHX) and cetylpyridinium chloride (CPC) are all known to inhibit production of volatile sulfur compounds (VSCs). The objective was to examine the anti-VSC dose-response effects of each of the above agents. Oral malodor was induced in 13 test subjects using the cysteine challenge method. The oral VSC response to rinses with 6 mm l-cysteine (pH 7.2) before and 1, 2 and 3 h after rinsing with zinc ions (Zn2+: 0.1, 0.3 and 1.0%), CHX and CPC (0.025 and 0.2%) was measured. Mouth air was analysed for VSC by gas chromatography (GC) according to current methodology. Zinc had a marked dose- and time-dependent anti-VSC effect. Zinc at 1% concentration had a somewhat unpleasant taste, whereas the lowest concentration was found acceptable. Chlorhexidine maintained a moderate anti-VSC effect over time. At 3 h, 0.2% CHX was the most effective agent but tasted relatively unpleasant. Cetylpyridinium at a concentration of 0.2% was only marginally more effective than 0.025% CHX over the 3 h, while 0.025% CPC had no better anti-VSC effect than water at both 2 h and 3 h. It was concluded that the three test agents demonstrated different anti-VSC kinetics. Although Zn had the best anti-VSC effect at 1 h, 0.2% CHX was at least as effective as 1% Zn at 3 h, most likely as a result of its unique substantivity.
This study evaluated the effectiveness of a newly developed dental hygiene chew for dogs, with and without a natural antimicrobial additive, compared with a reference diet. Efficacy was determined by measuring the severity of gingivitis and the accumulation of dental plaque and calculus in dogs after 4-weeks of being fed the different dietary regimens. Dogs fed a single daily dental chew had significantly less gingivitis (P = 0. 02), plaque (P = 0. 0004), and calculus (P = 0.0001) compared with dogs in the control group that were fed an identical diet but received no chews. The inclusion of the antimicrobial agent did not improve the efficacy of the product. The dental hygiene chews tested in this study have potential to help reduce the incidence of periodontal disease in dogs.
Dental plaque samples from (i) subjects with no apparent oral disease, (ii) mentally retarded subjects with periodontal disease, and (iii) subjects with active caries were collected in three transport media viz. a dithiothreitol poised balanced mineral salt solution designated as reduced transport fluid (RTF), VMG II, and modified Stuart medium (SBL). The samples were dispersed by sonic treatment, diluted in the respective medium in which they were collected, and cultured on MM10 sucrose agar. The efficiency of the transport media in the survival of dental plaque flora was determined by comparing the quantitative recovery (expressed as percentage of the initial viable count) from the specimens stored for various lengths of time. The data showed a great variation in the recovery of the oral bacterial flora from the plaque samples. VMG II and SBL served better than RTF as storage media for non-disease-associated dental plaque cultured under strict anaerobic conditions. Recoveries of bacteria from periodontal plaque specimens stored in RTF were higher than SBL and VMG II under identical conditions. The organisms present in the carious plaque samples appeared to survive much better in RTF and VMG II than in SBL as determined by conventional anaerobic culturing technique. However, VMG II showed a higher recovery of organisms from these specimens with an increase in the storage period, suggesting multiplication of the plaque flora. RTF did not allow the growth of oral bacterial flora under all experimental conditions. On the basis of the relative performance of these media it is suggested that RTF is a satisfactory medium for the transport of oral bacteria present in the samples.
It has been recognized for some time that bacterial species exist in complexes in subgingival plaque. The purpose of the present investigation was to attempt to define such communities using data from large numbers of plaque samples and different clustering and ordination techniques. Subgingival plaque samples were taken from the mesial aspect of each tooth in 185 subjects (mean age 51 +/- 16 years) with (n = 160) or without (n = 25) periodontitis. The presence and levels of 40 subgingival taxa were determined in 13,261 plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments were made at 6 sites per tooth at each visit. Similarities between pairs of species were computed using phi coefficients and species clustered using an averaged unweighted linkage sort. Community ordination was performed using principal components analysis and correspondence analysis. 5 major complexes were consistently observed using any of the analytical methods. One complex consisted of the tightly related group: Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. The 2nd complex consisted of a tightly related core group including members of the Fusobacterium nucleatum/periodonticum subspecies, Prevotella intermedia, Prevotella nigrescens and Peptostreptococcus micros. Species associated with this group included: Eubacterium nodatum, Campylobacter rectus, Campylobacter showae, Streptococcus constellatus and Campylobacter gracilis. The 3rd complex consisted of Streptococcus sanguis, S. oralis, S. mitis, S. gordonii and S. intermedius. The 4th complex was comprised of 3 Capnocytophaga species, Campylobacter concisus, Eikenella corrodens and Actinobacillus actinomycetemcomitans serotype a. The 5th complex consisted of Veillonella parvula and Actinomyces odontolyticus. A. actinomycetemcomitans serotype b, Selenomonas noxia and Actinomyces naeslundii genospecies 2 (A. viscosus) were outliers with little relation to each other and the 5 major complexes. The 1st complex related strikingly to clinical measures of periodontal disease particularly pocket depth and bleeding on probing.
Red complex bacteria (Treponema denticola, Tannerella forsythia and Porphyromonas gingivalis) play a major role in the aetiology of periodontal disease in humans. This study was designed to evaluate the association of such bacteria with periodontal disease in dogs.
Seventy-three subgingival samples taken from dogs ranging from 2 months to 12 years (median age 4 years) were tested for red complex bacteria using a polymerase chain reaction assay.
Thirty-six of 73 (49 · 3%) dogs were found to be positive for T. forsythia and P. gingivalis. Dogs with gingivitis or periodontitis were more likely to be infected with T. forsythia and P. gingivalis [odds ratio (OR) 5 · 4 (confidence interval (CI) 1 · 9-15 · 6), P = 0 · 002] than healthy animals. Only 3 (4 · 1%) of 73 samples were positive for red complex bacteria, but the association with periodontal disease was not significant. Conclusion And Clinical Relevance The results indicate that involvement of red complex bacteria in periodontal disease in dogs is similar to that observed in humans. Only the concurrent presence of T. forsythia and P. gingivalis were correlated to periodontal disease in dogs in this study.
An independent study was conducted to determine and quantify the oral care benefits of a daily edible dental chew in dogs as measured by plaque and calculus control, gingival indices, and oral malodor. A "clean mouth" test model was used comparing a commercial dry diet and a commercial dry diet plus one dental chew per day. The dental chew tested was representative of a retail canine dental chew. The test dental chew was a green-colored dental dog chew with a flexible texture that can be readily chewed by dogs. They are made with a knuckle bone shape on one end and a toothbrush shape on the other end. Sixty adult dogs were allocated in either control or test groups based on plaque stratification and studied for 28-days. The test group (30 dogs) received a dry diet and 1 dental chew each day. The control group (30 dogs) received the same dry diet only. At the end of the study, measurements of plaque and calculus accumulation and evaluations of oral malodor and gingival heath were performed. Adding a dental chew to the diet resulted in statistically significant reductions in plaque and calculus accumulation, and oral malodor while improving gingival indices.
Background and objectives:
Differentiation of live and dead cells is an important challenge when using molecular diagnosis for microbial identification. This is particularly relevant when bacteria have been exposed to antimicrobial agents. The objective of this study was to test a method using quantitative real-time polymerase chain reaction (qPCR) combined with propidium monoazide (PMA), developed for the selective quantification of viable P. gingivalis, A. actinomycetemcomitans, F. nucleatum and total bacteria in an in vitro biofilm model after antimicrobial treatment.
Material and methods:
PMA-qPCR method was tested in an in vitro biofilm model, using isopropyl alcohol as the antimicrobial agent. Matured biofilms were exposed for 1, 5, 10 and 30 min to isopropyl alcohol by immersion. Biofilms were disrupted and PMA added (final concentration of 100 μm). After DNA isolation, qPCR was carried out using specific primers and probes for the target bacteria. The differentiation of live and dead cells was tested by analysis of variance.
Results:
When PMA was used in the presence of viable target bacterial cells, no statistically significant inhibition of qPCR amplification was detected (p > 0.05 in all cases). Conversely, after immersion in isopropyl alcohol of the biofilm, PMA resulted in a significant total reduction of qPCR amplification of about 4 log10 . P. gingivalis showed a vitality reduction in the biofilm of 3 log10 , while A. actinomycetemcomitans and F. nucleatum showed a 2 log10 reduction.
Conclusion:
These results demonstrate the efficiency of PMA for differentiating viable and dead P. gingivalis, A. actinomycetemcomitans and F. nucleatum cells, as well as total bacteria, in an in vitro biofilm model, after being exposed to an antimicrobial agent. Hence, this PMA-qPCR method may be useful for studying the effect of antimicrobial agents aimed at oral biofilms.
Background and objectives:
One of the major disadvantages of DNA-based microbial diagnostics is their inability to differentiate DNA between viable and dead microorganisms, which could be important when studying etiologically relevant pathogens. The aim of this investigation was to optimize a method for the selective detection and quantification of only viable Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis cells by combining quantitative real-time polymerase chain reaction (qPCR) and propidium monoazide (PMA).
Material and methods:
Three different concentrations of PMA (10, 50 or 100 μm) were added to suspensions of 10(6) (CFU)/mL of viable/dead A. actinomycetemcomitans and P. gingivalis cells. After DNA isolation, qPCR was carried out using specific primers and probes for the tested bacteria. PMA was further tested with different mixtures containing varying ratios of viable and dead cells. The efficacy of PMA to detect viable/dead cells was tested by analysis of variance.
Results:
For these specific bacterial pathogens, 100 μm PMA resulted in a significant reduction of qPCR amplification with dead cells (10(6) CFU/mL), while with viable cells no significant inhibition was detected. PMA was also effective in detecting selectively viable cells by qPCR detection, when mixtures of varying ratios of viable and dead bacteria were used.
Conclusions:
This study demonstrated the efficiency of PMA for differentiating viable and dead A. actinomycetemcomitans and P. gingivalis cells. This method of PMA-qPCR may be useful for monitoring new antimicrobial strategies and for assessing the pathogenic potential of A. actinomycetemcomitans and P. gingivalis in different oral conditions when using molecular diagnostic methods.
The type and frequency of dental diseases and disorders were determined in 162 randomly selected dogs available for necropsy. There were 82 males and 80 females ranging in age from seven months to 14 years. There were 150 purebred dogs (50 different breeds) and 12 mongrels. Only four breeds were represented by ten individuals or more: Poodles (Miniature and Toy), German Shepherds, Dachshunds, and Boxers. The oral cavity was examined macroscopically, and missing teeth, dental caries, dental calculus (tartar), and gingival epulides were recorded as to frequency and distribution. After macroscopic examination of the oral cavity, the head of each dog was sawed at the midline, fixed in formalin, and radiographed. The frequency, severity, and distribution of marginal periodontitis, periapical destruction, and root resorption were recorded. The most frequently noted disease was periodontitis, which increased in frequency and severity with increasing age of the dogs. The disease varied markedly among and within different breeds. Small dogs were more often affected with periodontitis than large dogs. Periapical destruction was a common finding. Caries and root resorption were less common. Missing teeth was a frequent disorder regardless of age. Most of the dogs had dental calculus.
Periodontal disease is an oral disease common in middle-aged dogs and cats, with halitosis being the most common sign. There
are many commercial products containing chlorhexidine, cetylpyridinium chloride (CPC), zinc salts and essential oils for controlling
halitosis and periodontal disease. CPC is a quaternary ammonium compound and has a broad spectrum antimicrobial activity.
In this study, oral spray (OS) and gel (OG) containing CPC was applied to the dogs’ teeth twice daily for 3 weeks, and their
effect in controlling periodontal disease and halitosis was examined. In the 3-week study, OS and OG were significantly effective
in controlling plaque, calculus, and halitosis. Therefore, the OS and OG containing CPC were effective in controlling periodontal
disease and halitosis in dogs.
To evaluate the efficacy of a novel toothpaste containing zinc ions and o-cymen-5-ol to reduce volatile sulfur compounds (VSCs) in in vitro models and to elucidate the mode of action for any activity observed.
Three models were employed, a chemical neutralisation model to evaluate the chemical reactivity of toothpaste slurries to VSCs, a biofilm perfusion model to measure activity in an orally-relevant biofilm and a planktonic bacterial model to measure antimicrobial effects.
The models showed that zinc ions were able to react chemically with hydrogen sulfide to remove this odorous component of halitotic breath. This activity was confirmed within a complex biofilm model, with over 90% of hydrogen sulfide removed from perfusate gas by a slurry of the test toothpaste.
This work provides a mode of action for the clinically observed reduction in VSCs seen for up to 12 hours post brushing with this novel toothpaste.
Sixteen toy breed dogs completed a parallel, 70-day two-period, cross-over design clinical study to determine the effect of a vegetable dental chew on gingivitis, halitosis, plaque, and calculus accumulations. The dogs were randomly assigned into two groups. During one study period the dogs were fed a non-dental dry diet only and during the second study period were fed the same dry diet supplemented by the daily addition of a vegetable dental chew. Daily administration of the dental chew was shown to reduce halitosis, as well as, significantly reduce gingivitis, plaque and calculus accumulation and therefore may play a significant role in the improvement of canine oral health over the long-term.
The aim of this study was to evaluate the effects of ZnCl(2) on plaque-growth and vitality pattern of dental biofilm and to determine the optimum zinc concentration for the inhibition of plaque formation.
Data were collected from nine volunteers for whom a special-designed acrylic appliance was prescribed after a careful dental check up. The volunteers rinsed twice daily for 2min with ZnCl(2) of 2.5, 5, 10, 20mM as treatment and double distilled water (DDW) as control in respective assigned test weeks. The plaque index (PI) was assessed after 48h of appliance wearing. The glass discs with the adhered biofilm were removed from the splints and stained with two fluorescent dyes. The biofilm thickness (BT) and bacterial vitality of the whole biofilm as well as the mean bacterial vitality (BV) of the inner, middle and outer layers of biofilm were evaluated under confocal laser scanning microscope (CLSM).
PI, BT and BV of biofilms treated by various concentrations of ZnCl(2) were reduced significantly when compared with the DDW group (p<0.05). PI, BT and BV of the 2.5mM ZnCl(2) group was significantly higher than groups of 5, 10, 20mM ZnCl(2) (p<0.05). The mean BV of the 3 layers (inner, middle and outer layers) showed that 2.5mM ZnCl(2) was the lowest concentration to inhibit BV in the outer layer, 5mM was the lowest concentration to extend this inhibition of BV to the middle layer, and none of the concentrations investigated in this study has shown any effect on bacteria inhibition in the inner layer.
Zinc ions exhibited possible inhibitory effects on plaque formation, and have a promising potential to be used as an antibacterial agent in future dentifrices and mouthrinses.
The loads and locations of bacterial types associated with oral malodour on the tongue surface and gingival crevice were investigated in man and the dog respectively. In the human study, samples were taken from 50 subjects with brushes at the dorsal anterior, dorsal middle, dorsal posterior, dorsal posterior to the circumvallate papillae (DPCP), lateral posterior and ventral posterior (VP) surfaces, and cultured appropriately. Malodour was assessed by trained judges. Mean volatile sulfur compound (VSC) producing bacterial counts (colony forming units/brush × 10(5)) were found to be highest (88.94) and lowest (0.33) at the DPCP and VP sites respectively. Anaerobic, gram-negative and VSC counts at DPCP surfaces increased with malodour intensity, whereas aerobic and S. salivarius counts decreased. The prevalence and populations of the VSC producing Porphyromonas and Prevotella species were determined in the dental plaque from 34 dogs. Porphyromonas gulae and Prevotella intermedia were present in 68% and 44% of dogs, and 47% and 23% of plaque samples respectively. P. gulae and Prev. intermedia counts increased with plaque quantity (P < 0.05) and gingivitis (P < 0.1). The close association observed between canine periodontal disease and measurements of oral malodour is supported.
Naturally occurring periodontal disease affects >75% of dogs and has been associated with cardiac lesions and presumptive endocarditis. However, the relationships between periodontal disease and chronic kidney disease (CKD) in dogs have not been studied. In a retrospective longitudinal study the incidence of azotemic CKD was compared between a cohort of 164,706 dogs with periodontal disease and a cohort of age-matched dogs with no periodontal disease from a national primary care practice. These dogs contributed 415,971 dog-years of follow-up from 2002 to 2008. Hazard ratios and 95% confidence intervals from Cox regression were used to compare the incidence of azotemic CKD in dogs with stage 1, 2, or 3/4 periodontal disease to dogs with no periodontal disease. The hazard ratio for azotemic CKD increased with increasing severity of periodontal disease (stage 1 hazard ratio=1.8, 95% confidence interval: 1.6, 2.1; stage 2 hazard ratio=2.0, 95% confidence interval: 1.7, 2.3; stage 3/4 hazard ratio=2.7, 95% confidence interval: 2.3, 3.0; P(trend)=<0.0001) after adjustment for age, gender, neuter status, breed, body weight, number of hospital visits, and dental procedures. Increasing severity of periodontal disease was also associated with serum creatinine >1.4 mg/dl and blood urea nitrogen >36 mg/dl, independent of a veterinarian's clinical diagnosis of CKD.
There are few in vitro models available in the scientific literature for study of the structure, formation and development of the subgingival biofilm. The purpose of this study was to develop and validate an in vitro biofilm model, using representative selected bacteria from the subgingival microbiota.
Six standard reference strains were used to develop biofilms over sterile ceramic calcium hydroxyapatite discs coated with saliva within the wells of presterilized polystyrene tissue culture plates. The selected species represent initial (Streptococcus oralis and Actinomyces naeslundii), early (Veillonella parvula), secondary (Fusobacterium nucleatum) and late colonizers (Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans). The structure of the biofilm obtained was studied using a vital fluorescence technique in conjunction with confocal laser scanning microscopy. The biofilm bacterial kinetics were studied by terminal restriction fragment length polymorphism analysis.
After 12 h, initial and early colonizers were the first microorganisms detected adhering to the calcium hydroxyapatite discs. The intermediate colonizer F. nucleatum was not detected in the model until 24 h of incubation. Late colonizers A. actinomycetemcomitans and P. gingivalis could be measured inside the biofilm after 48 h. The biofilm reached its steady state between 72 and 96 h after inoculation, with bacterial vitality increasing from the hydroxyapatite surface to the central part of the biofilm.
An in vitro biofilm model was developed and validated, demonstrating a pattern of bacterial colonization and maturation similar to the in vivo development of the subgingival biofilm.
This clinical investigation was conducted to evaluate the impact of a unique soft rawhide product on the formation of dental calculus in a housed colony of beagle dogs. The study was designed as a 2-way crossover test with 4-week test periods and a 1-week wash-out period between the test periods. All dogs were provided a dry commercial diet and tap water ad libitum. The dogs were stratified into two groups of matched pairs on the basis of their calculus and plaque formation rates. Each test period was initiated immediately following a complete supragingival scaling and crown polishing and the dogs in the test group received a soft rawhide treat about 4-hours after their normal feeding. Examinations were performed about 20 to 24-hours after the last feeding by an experienced clinical examiner using conventional indices. The data indicated that once daily feeding of the soft rawhide chew product resulted in statistically significant reductions in the formation of dental calculus (28.0%), dental plaque (19.0%), and gingivitis (46.0%).
To test the hypothesis that increased severity of periodontal disease in dogs is associated with an increased risk of cardiovascular-related events, such as endocarditis and cardiomyopathy, as well as markers of inflammation.
Historical cohort observational study.
59,296 dogs with a history of periodontal disease (periodontal cohort), of which 23,043 had stage 1 disease, 20,732 had stage 2 disease, and 15,521 had stage 3 disease; and an age-matched comparison group of 59,296 dogs with no history of periodontal disease (nonperiodontal cohort).
Cox proportional hazard regression models were used to estimate the risk of cardiovascular-related diagnoses and examination findings in dogs as a function of the stage of periodontal disease (1, 2, or 3 or no periodontal disease) over time while controlling for the effect of potential confounding factors.
Significant associations were detected between the severity of periodontal disease and the subsequent risk of cardiovascular-related conditions, such as endocarditis and cardiomyopathy, but not between the severity of periodontal disease and the risk of a variety of other common noncardiovascular-related conditions.
The findings of this observational study, similar to epidemiologic studies in humans, suggested that periodontal disease was associated with cardiovascular-related conditions, such as endocarditis and cardiomyopathy. Chronic inflammation is probably an important mechanism connecting bacterial flora in the oral cavity of dogs with systemic disease. Canine health may be improved if veterinarians and pet owners place a higher priority on routine dental care.
To evaluate the anti-malodor efficacy of toothpaste containing 0.2% zinc sulphate after a single brushing and following extended use.
Two clinical studies of common design but different malodor assessment were carried out. In the first study, volatile sulphur compounds (VSC) were measured with a Halimeter; in the second, breath freshness was assessed by odor judges (organoleptic score). At the start of the study, suitable subjects were assigned at random to either the test toothpaste (containing 0.2% zinc sulphate) or the placebo toothpaste (no zinc), and oral breath assessments were made before and two hours after brushing. The same assessments were repeated after allocated toothpastes had been used twice daily for four weeks. Prior to the before-brushing breath assessment, subjects were asked to refrain from eating/drinking and oral hygiene for 12 hours.
The results at the start and end of the study showed a significant reduction in mean Halimeter scores and a significant improvement in organoleptic scores two hours after brushing for subjects using the test toothpaste with zinc sulphate. In addition, at the end of the study, subjects using the test toothpaste with zinc sulphate had reduced Halimeter scores and improved organoleptic scores 12 hours after the last brushing compared to the placebo group. The differences between the test and placebo values ranged from 16% to 46% and were all statistically significant (p < 0.0001).
Both clinical studies independently support the efficacy of the test toothpaste with 0.2% zinc sulphate in reducing oral malodor after a single brushing, and after four weeks for 12 hours after brushing compared to a toothpaste without zinc ions.
Dental plaque samples from (i) subjects with no apparent oral disease, (ii) mentally retarded subjects with periodontal disease, and (iii) subjects with active caries were collected in three transport media viz. a dithiothreitol poised balanced mineral salt solution designated as reduced transport fluid (RTF), VMG II, and modified Stuart medium (SBL). The samples were dispersed by sonic treatment, diluted in the respective medium in which they were collected, and cultured on MM10 sucrose agar. The efficiency of the transport media in the survival of dental plaque flora was determined by comparing the quantitative recovery (expressed as percentage of the initial viable count) from the specimens stored for various lengths of time. The data showed a great variation in the recovery of the oral bacterial flora from the plaque samples. VMG II and SBL served better than RTF as storage media for non-disease-associated dental plaque cultured under strict anaerobic conditions. Recoveries of bacteria from periodontal plaque specimens stored in RTF were higher than SBL and VMG II under identical conditions. The organisms present in the carious plaque samples appeared to survive much better in RTF and VMG II than in SBL as determined by conventional anaerobic culturing technique. However, VMG II showed a higher recovery of organisms from these specimens with an increase in the storage period, suggesting multiplication of the plaque flora. RTF did not allow the growth of oral bacterial flora under all experimental conditions. On the basis of the relative performance of these media it is suggested that RTF is a satisfactory medium for the transport of oral bacteria present in the samples.
Subjects using 30 mM zinc phenolsulphonate as a mouthwash retained 12 per cent of the zinc. Salivary zinc concentration was increased by using mouthwashes containing 17-35 mM zinc as the sulphate, phenolsulphonate or citrate. For 17 mM zinc sulphate or phenolsulphonate, the effect lasted 3-4 h. Zinc retained in the mouth gave visible fluorescence after rinsing with 8-hydroxyquinoline and was particularly evident on the tongue, cheek mucosa and dental plaque. The concentration of zinc in plaque was increased 13-19-fold 1 h after using 31 or 18 mM zinc phenolsulphonate. A 3-fold increase was still present 6 h later for the 31 mM mouthwash. Zinc salts inhibited acid production from [14C]-glucose in vitro by plaque at concentrations which were found in plaque in vivo after using the mouthwashes. The effect of zinc on the metabolic activity of plaque may reduce the growth rate of plaque-bacteria and so decrease plaque growth.
The prevalence and populations of Porphyromonas and Prevotella species were determined in three samples of dental plaque from each of 34 healthy dogs. Porphyromonas gingivalis was present in 68 per cent of the dogs and 47 per cent of the plaque samples. The counts of P gingivalis increased with age (P < 0.1), the amount of plaque (P < 0.05) and the degree of gingivitis (P < 0.1). Prevotella intermedia was present in 44 per cent of the dogs and 23 per cent of the plaque samples. The counts of P intermedia were correlated with the amount of plaque (P < 0.05) and the degree of gingivitis (P < 0.1). Porphyromonas canoris, P salivosa, P cangingivalis, P cansulci, P crevioricanis and Prevotella denticola were isolated from only 9, 6, 3, 3, 3 and 3 per cent of the dogs, respectively. Porphyromonas gingivicanis was not isolated from any of the animals sampled. There was a highly significant (P < 0.001) correlation between the amount of plaque, the extent of gingivitis and the age of the dog.
It has been recognized for some time that bacterial species exist in complexes in subgingival plaque. The purpose of the present investigation was to attempt to define such communities using data from large numbers of plaque samples and different clustering and ordination techniques. Subgingival plaque samples were taken from the mesial aspect of each tooth in 185 subjects (mean age 51 +/- 16 years) with (n = 160) or without (n = 25) periodontitis. The presence and levels of 40 subgingival taxa were determined in 13,261 plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments were made at 6 sites per tooth at each visit. Similarities between pairs of species were computed using phi coefficients and species clustered using an averaged unweighted linkage sort. Community ordination was performed using principal components analysis and correspondence analysis. 5 major complexes were consistently observed using any of the analytical methods. One complex consisted of the tightly related group: Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. The 2nd complex consisted of a tightly related core group including members of the Fusobacterium nucleatum/periodonticum subspecies, Prevotella intermedia, Prevotella nigrescens and Peptostreptococcus micros. Species associated with this group included: Eubacterium nodatum, Campylobacter rectus, Campylobacter showae, Streptococcus constellatus and Campylobacter gracilis. The 3rd complex consisted of Streptococcus sanguis, S. oralis, S. mitis, S. gordonii and S. intermedius. The 4th complex was comprised of 3 Capnocytophaga species, Campylobacter concisus, Eikenella corrodens and Actinobacillus actinomycetemcomitans serotype a. The 5th complex consisted of Veillonella parvula and Actinomyces odontolyticus. A. actinomycetemcomitans serotype b, Selenomonas noxia and Actinomyces naeslundii genospecies 2 (A. viscosus) were outliers with little relation to each other and the 5 major complexes. The 1st complex related strikingly to clinical measures of periodontal disease particularly pocket depth and bleeding on probing.
In 28 dogs, oral malodor was assessed organoleptically (0-3 scale) and by measurement of volatile sulfur components (VSC), using two positions ('intraoral' and 'tooth surface') for sampling VSC. Significant correlations were found between: intraoral and tooth surface VSC collection positions (p < 0.0001) and between organoleptic and tooth surface VSC data (p < 0.0001). VSC measurement is a sensitive, repeatable and non-subjective method of assessing oral malodor in dogs.
A study to compare the effect of a dental hygiene chew with or without 0.2% chlorhexidine on the development of gingivitis and the accumulation of dental deposits was performed using 11 small dogs. Confirming previous data, the daily addition of a standard chew to a dry diet resulted in significantly less gingivitis and calculus after 3 weeks compared with feeding the dry diet alone. Addition of chlorhexidine to the chew made no difference to the degree of gingivitis or the amount of calculus that accumulated, but did result in significantly less plaque accumulation after 3 weeks. The abrasiveness of the chew, rather than the antibacterial activity of chlorhexidine, is likely to have contributed the most to the maintenance of oral health in dogs with mild gingivitis.
The objective of the study was to investigate the long-term effects of feeding a dental hygiene chew that has been shown to be effective in promoting periodontal health in dogs in short-term studies. Oral malodor, calculus, and plaque scores were still significantly lower after 21 months in the group that was receiving the dental hygiene chew, although gingivitis scores no longer differed significantly. There were no reports of any adverse reactions during the study. The results of the study support that feeding of the dental hygiene chew six days per week reduces accumulation of dental deposits, helps maintain periodontal health, and increases the time interval between professional periodontal intervention.
Tooth brushing is considered a superior technique for reducing plaque accumulation. Other methods of maintaining oral hygiene have been investigated since many owners may not be willing or able to brush their dog's teeth. Following a professional teeth cleaning procedure, 11 dogs were offered a rawhide dental chew BID for 7-days, while 11 other dogs were fed the same diet without receiving the chew device. Dogs in the treatment group had significantly less plaque formation during the trial period compared with dogs in the control group. The rawhide dental chew provided in the study reported here decreases plaque formation in the short-term and may be beneficial in the prevention of progressive periodontal disease associated with attachment loss if provided on a long-term basis.
The clinical and microbiological effects of zinc ascorbate gel applied orally in cats were evaluated during a 42-day study period. Cats were divided randomly into two equal groups, with the treatment group (18 cats) receiving zinc ascorbate gel and the control group (18 cats) receiving a placebo (0.9% sterile saline). Clinical parameters evaluated biweekly included halitosis, plaque, calculus, and gingivitis. Aerobic and anaerobic bacterial cultures were obtained from cats in the treatment group at Days 0 and 42. There was a significant decrease in plaque, gingivitis, and anaerobic periodontal pathogens in treatment group cats. Halitosis and calculus scores were not significantly different in treatment group compared with control group cats. The results of this study suggest that zinc ascorbate gel used as an oral antiseptic improves feline oral health, and may be most effective in decreasing bacterial growth, plaque formation, and gingivitis when applied following a professional teeth cleaning procedure.
To evaluate the in vitro and in vivo antimicrobial activity of four commercial 0.12% chlorhexidine mouthrinses.
The in vitro antimicrobial activity test consisted in a modified contact test where 20 selected bacterial species were tested during 1 min with each test product. After the contact, the inoculum was cultured, and the results were expressed in terms of survival/resistance and the percentage of survival as compared to a saline control. The in vivo test consisted of a double-blind, randomized, crossover salivary bacterial counts study. 10 volunteers rinsed during 1 min with each tested product. Saliva samples were obtained before rinsing, and after 5 min, and 1, 3, 5 and 7 h. These samples were cultured both aerobically and anaerobically. Percentages of survival, in regard to baseline, were calculated for each time point. Comparisons among products were tested using anova and selected paired t-test.
The in vitro contact test showed no survival in any tested species with CHX+CPC, while three species (Lactobacillus casei, Streptococcus mitis and Peptostreptococcus micros) were resistant to the other three products. CHX and CHX+NaF demonstrated additional resistant species (three and four species, respectively). The in vivo salivary bacterial counts test showed higher reductions of CHX+CPC and CHX+ALC in aerobic and anaerobic bacteria, lasting for 5 h. Significant differences were detected at multiple time points, when these two products were compared both with the control and the other tested products.
Important differences in activity, among 0.12% CHX products, were detected by both in vitro and in vivo tests. The formulation with alcohol was more active than those without alcohol, excepting the formulation with CHX+CPC, in which the reformulation and addition of CPC not only compensate but rather increase the antimicrobial activity.
The etiology of human periodontal disease has been the focus of considerable research, yet relatively little is known about the causative agents of companion animal periodontitis. In humans, Porphyromonas gingivalis, a black-pigmented anaerobic bacteria (BPAB), has been implicated as the primary periopathogen. It has been demonstrated that BPAB are also found in companion animal periodontal pockets. While some animal BPAB have been individually identified, a study to identify the most frequently isolated subgingival BPAB has not been completed using genetic tools. The objective of this work was to identify the types and relative frequencies of pigmented anaerobic bacteria found in the periodontal pockets of dogs. Porphyromonas salivosa, Porphyromonas denticanis (a novel species) and Porphyromonas gulae were found to be the most frequently isolated BPAB associated with canine periodontitis.
The purpose of this clinical cross-over study was to examine the antibacterial and plaque-inhibiting properties of two chlorhexidine solutions compared with a negative control.
Twenty-one volunteers refrained from all oral hygiene measures, but rinsed instead twice daily with 10 ml of a conventional chlorhexidine solution (0.2%; CHX), a chlorhexidine solution with anti-discolouration system (ADS) (0.2%, alcohol-free chlorhexidine solution (CSP)) or a placebo solution (Pla). Plaque index (PI), plaque area (PA) and bacterial vitality were assessed after 24 h (PI1, vital flora (VF)1) and 96 h (PI2; VF2, PA). After a 10-day wash-out period, a new test cycle was started.
Results for Pla were 0.94, 1.59, 27.4 (PI1, PI2, PA) and 79% and 72% (VF1 and VF2). CSP significantly reduced the parameter PI1, PI2 and PA to 0.67 (p=0.012), 1.0 and 15.7 (p<0.001). VF1 and VF2 (63% and 53%) were not significantly affected. The corresponding figures of CHX were 0.42, 0.43, 6.77, 33 and 16%, which were all significantly lower (all p<0.001). On comparing the two chlorhexidine solutions, CHX showed significantly higher reductions of all parameters.
The results suggest that the 0.2% alcohol-containing solution showed superiority in inhibiting plaque re-growth and reducing bacterial vitality compared with the solution with ADS.
Over a period of 4-months, feeding small breed dogs with a regular dry diet supplemented with a daily oral hygiene chew resulted in a significant reduction in plaque deposition (17.3%) and calculus accumulation (45.8%) compared with dogs fed the dry diet alone. Daily feeding of a specifically designed oral hygiene chew that reduces plaque and calculus accumulation may play a role in the severity of canine periodontal disease.
Porphyromonas gulae is black-pigmented anaerobic bacteria associated with canine periodontitis. There is little information available about the specific identify and relative occurrence of pigmented anaerobes in companion animals. Our aim was to clarify the factor involved in the adherence and colonization of the organism in the oral cavity. Fimbrial protein was purified from P. gulae ATCC 51700. The molecular mass of this protein was approximately 41kDa as estimated by SDS-PAGE. An antibody against 41-kDa fimbrial protein from P. gingivalis ATCC 33277 reacted with fimbrillin of P. gulae ATCC 51700. Immunogold electron microscopy revealed that the anti-41kDa fimbrial serum bound to fimbria on the cell surface of P. gulae ATCC 51700. Thus, fimbrial protein of P. gulae ATCC 51700 had the same size and antigenicity as 41-kDa fimbriae of P. gingivalis ATCC 33277. The nucleotide sequence of the fimA gene from P. gulae ATCC 51700 showed 94% homology with that of P. gingivalis ATCC 33277. Moreover, the deduced amino acid sequences have 96.8% identity. P. gulae has adherent ability to gingival epithelial cells. The properties of P. gulae fimbriae are similar to those of P. gingivalis fimbriae. We suggest that the surface structure of P. gulae may play a role in the colonization of this organism in periodontal pockets in companion animals.
Differences in antimicrobial activity of four commercial 0.12% chlorhexidine mouthrinse formulations: an in vitro contact test and salivary bacterial counts study
- D Herrera
- S Roldán
- Santa Cruz
Herrera D, Roldán S, Santa Cruz I, et al. Differences in antimicrobial activity of four commercial 0.12% chlorhexidine
mouthrinse formulations: an in vitro contact test and salivary
bacterial counts study. J Clin Periodontol 2003;30:307–314.
A macroscopic and radiological investigation of dental diseases of the dog
- S E Hamp
- S E Ollon
- K Farso-Madsen
Hamp SE, Ollon SE, Farso-Madsen K, et al. A macroscopic and
radiological investigation of dental diseases of the dog. Vet Radiol 1984;25:86-92.
A macroscopic and radiological investigation of dental diseases of the dog
- Hamp