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1.  “Alas poor Yorick”: What retrospective analysis of canine skulls can tell us about the impact of environmental factors on health 
Necropsies and extensive histological evaluation for clinical and sub-clinical disease of approximately three hundred Portuguese Water dogs are available as part of an ongoing study to assess their state of health at end of life. Throughout life these dogs enjoyed a variety of lifestyles and environments. Here we carry out retrospective quantitative assessments of life-time dietary input and physical activity for each dog. To do this, collagens from skull vault bone and from dentine have been analyzed for ratios of stable isotopes to determine differences in diet that individual dogs experienced during late or early life respectively. Robustness of skull bone (weight/unit of skull size) was used as a relative indicator of the amount of physical activity experienced during a dog’s lifetime. These environmental parameters were correlated with the frequency and severity of specific disease processes determined at necropsy. Both measures were shown to exert significant low-level (r < 25%) differential effects on specific diseases. The value of retrospective analysis of environmental influences is discussed.
PMCID: PMC3891401  PMID: 24436781
Portuguese Water Dog; Necropsy; Histopathology; Exercise; Nutrition; Stable Isotope; Skull; Dentine; Collagen
2.  Cervical lung lobe herniation in dogs identified by fluoroscopy 
The Canadian Veterinary Journal  2013;54(10):955-959.
This study aimed to determine the frequency of cervical lung lobe herniation (CLLH) in dogs evaluated fluoroscopically and to identify associated characteristics. Reports of diagnostic procedures and patient summaries from 2008 to 2010 were reviewed retrospectively. Signalment, body weight, duration of cough, presence of heart murmur and airway collapse, and radiographic findings were compared between dogs with and without CLLH. Of the 121 dogs that were examined, CLLH occurred in 85 (70%). The extra-thoracic trachea kinked during herniation in 33 (39%) dogs with CLLH. Collapse of the intra-thoracic trachea (assessed fluoroscopically or bronchoscopically) and collapse of major bronchi (assessed fluoroscopically) were strongly associated with CLLH. Although redundant dorsal tracheal membrane on radiographs was associated with CLLH, extra-thoracic tracheal collapse, assessed fluoroscopically or bronchoscopically, was not. No other associations were found. Cervical lung lobe herniation was present in most dogs evaluated during cough and was associated with intra-thoracic large airway collapse, but not duration of cough.
PMCID: PMC3781426  PMID: 24155415
3.  Clinical and mycological analysis of dog’s oral cavity 
Brazilian Journal of Microbiology  2013;44(1):139-143.
The oral microbiota of humans and animals is made up of a wide variety of yeasts and bacteria, but microbiota of dogs is not totally described. Although such identification is an important step to establish the etiopathogenesis and adequate therapy for the periodontal disease The aim of this study was to evaluate and correlate oral alterations with the presence of yeasts in oral cavity of female dogs. After clinical evaluation samples from healthy and from dogs with oral diseases were obtained from three different oral sites by swabs, curettes, millimeter periodontal probes and HA membrane tip in cellulose ester. Yeast identification was performed through macroscopic and microscopic colony features and biochemical tests. Dental calculus was the most prevalent occurrence in the oral cavity of 59 females. However, the isolation of yeasts was significantly higher (p < 0.05) in animals suffering from halitosis. Eleven yeast species were identified, namely: Malassezia pachydermatis, Rhodotorula spp., Candida albicans, C. catenulata, C. famata, C. guilliermondii, C. parapsilosis, C. intermedia, Trichosporon asahii, T. mucoides and Cryptococcus albidus. It could be concluded that the yeasts are part of the microbiota from the different sites of the oral cavity of the female canines studied without causing any significant alterations except halitosis.
PMCID: PMC3804190  PMID: 24159296
oral microbiota; yeasts; M. pachydermatis. Candida spp.; halitosis
4.  Periodontal tissue reaction to customized nano-hydroxyapatite block scaffold in one-wall intrabony defect: a histologic study in dogs 
This study evaluated histologically the tissue responses to and the effects of a customized nano-hydroxyapatite (n-HA) block bone graft on periodontal regeneration in a one-wall periodontal-defect model.
A customized block bone for filling in the standardized periodontal defect was fabricated from prefabricated n-HA powders and a polymeric sponge. Bilateral 4×4×5 mm (buccolingual width×mesiodistal width×depth), one-wall, critical-size intrabony periodontal defects were surgically created at the mandibular second and fourth premolars of five Beagle dogs. In each dog, one defect was filled with block-type HA and the other served as a sham-surgery control. The animals were sacrificed following an 8-week healing interval for clinical and histological evaluations.
Although the sites that received an n-HA block showed minimal bone formation, the n-HA block was maintained within the defect with its original hexahedral shape. In addition, only a limited inflammatory reaction was observed at sites that received an n-HA block, which might have been due to the high stability of the customized block bone.
In the limitation of this study, customized n-HA block could provide a space for periodontal tissue engineering, with minimal inflammation.
PMCID: PMC3349047  PMID: 22586523
Bone substitutes; Guided tissue regeneration; Periodontal disease; Tissue engineering; Tissue scaffolds
5.  The histological appearance of peroral gastric biopsies in clinically healthy and vomiting dogs. 
A survey of the histology of gastric biopsies in 501 dogs, consisting of 19 clinically healthy dogs and 482 vomiting dogs is presented. Whole stomachs of four young clinically healthy laboratory dogs were used as controls. Eleven percent of forceps biopsies were unsuitable for examination; all suction biopsies were of good quality. Slight to severe gastritis was found in 168 vomiting dogs (35%), whereas five dogs (26%) of the clinically healthy group showed a mainly slight gastritis. Superficial and diffuse gastritis were the most prominent findings in the 168 dogs with gastritis. A single type of gastritis was found in 114 dogs, a combination of different types in 54 dogs. Gastric atrophy was seen in 23 (5%) vomiting dogs and in three (15%) clinically healthy dogs, atrophy with a slight to severe fibrosis in 21 (4%) vomiting dogs, and in 84 (17%) vomiting dogs and two (11%) healthy dogs, gastric fibrosis was present. Carcinomas were seen in 26 vomiting dogs, of which 17 also had gastritis. A differential diagnosis of granulomatous gastritis/carcinoma had to be made in one case. Seven dogs showed a lymphosarcoma, and in six other dogs a differential diagnosis of lymphosarcoma and/or gastritis was made. One adenomatous polyp was seen. In one clinically healthy dog an adenomyoma was diagnosed. Ulceration was found in 24 dogs, but only five of these lacked other lesions. Other biopsy findings were pseudopyloric metaplasia, hyperplasia, cysts, calcification and edema. Some dogs showed "antralization".
PMCID: PMC1255403  PMID: 3349402
6.  Histological Analysis of the Effect of Accelerated Portland Cement as a Bone Graft Substitute on Experimentally-Created Three-Walled Intrabony Defects in Dogs 
Background and aims
Recent literature shows that accelerated Portland cement (APC) is a non-toxic material that may have potential to promote bone healing. The objective of this study was to histologically evaluate periodontal healing focusing on new bone regeneration following implantation of APC into intra-bony defects in dogs.
Materials and methods
Three-wall intra-bony periodontal defects were surgically created at the mesial aspect of the first molar in both sides of mandible in six dogs. One side was randomly filled with the material and other received a flap operation only. The animals were euthanized eight weeks post-surgery when block sections of the defect sites were collected and prepared for qualitative histological analysis.
Compared to control group, stimulation of growth of new bone tissue in the cavity con-taining APC was significantly prominent in three of six cases, showing osteoid formation with osteoblastic rimming and new bone trabeculla. New bone formation was observed just close to cavity containing APC. Connective tissue proliferation and downgrowth of epithelium were signif-icantly less than those of control group.
Our results are encouraging for the use of APC as a bone substitute, but more comprehensive study are necessary before warranting clinical use.
PMCID: PMC3529889  PMID: 23277848
Accelerated Portland cement; bone substitutes; osteogenesis
7.  Unusual presentation of alveolar echinococcosis as prostatic and paraprostatic cysts in a dog 
Alveolar echinococcosis (AE) is caused by the larval stage (metacestode) of Echinococcus multilocularis. The domestic dog can act as a definitive host and harbor adult cestodes in its small intestine or become an aberrant intermediate host carrying larval stages that may cause severe lesions in the liver, lungs and other organs with clinical signs similar to AE in humans.
Case presentation
A case of canine AE, affecting the liver and prostate with development of multilocular hydatid paraprostatic cysts and possible lung involvement is described in an 8–year-old neutered male Labrador retriever dog.
The dog presented with progressive weight loss, acute constipation, stranguria and a suspected soft tissue mass in the sublumbar region. Further evaluation included computed tomography of the thorax and abdomen, which revealed cystic changes in the prostate, a paraprostatic cyst, as well as lesions in the liver and lungs. Cytological examination of fine-needle aspirates of the liver, prostate and paraprostatic cyst revealed parasitic hyaline membranes typical of an Echinococcus infection and the presence of E. multilocularis-DNA was confirmed by PCR.
The dog was treated with albendazole and debulking surgery was considered in case there was a good response to antiparasitic treatment. Constipation and stranguria resolved completely. Six months after the definitive diagnosis, the dog was euthanized due to treatment-resistant ascites and acute anorexia and lethargy.
To the authors’ knowledge, this is the first publication of an E. multilocularis infection in a dog causing prostatic and paraprostatic cysts. Although rare, E. multilocularis infection should be considered as an extended differential diagnosis in dogs presenting with prostatic and paraprostatic disease, especially in areas where E. multilocularis is endemic.
PMCID: PMC3765175  PMID: 23938212
Echinococcus multilocularis; Prostatic cyst; Paraprostatic cysts; Cytology; Radiology; PCR; Canine alveolar echinococcosis; Dog
8.  Histopathological and parasitological study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum 
The aim of this study was to provide a systematic pathological and parasitological overview of the gastrointestinal tract (GIT), including the stomach, duodenum, jejunum, ileum, caecum and colon, of dogs naturally infected with Leishmania.
Twenty mongrel dogs naturally infected with Leishmania (Leishmania) infantum and obtained from the Control Zoonosis Center of the Municipality of Ribeirão das Neves, Belo Horizonte Metropolitan area, Minas Gerais (MG) state, Brazil, were analyzed. The dogs were divided into two groups: Group 1 comprised nine clinically normal dogs and group 2 comprised 11 clinically affected dogs. After necropsy, one sample was collected from each GIT segment, namely the stomach, duodenum, jejunum, ileum, caecum and colon. Furthermore, paraffin-embedded samples were used for histological and parasitological (immunohistochemistry) evaluation and a morphometrical study were carried out to determine the parasite load (immunolabeled amastigote forms of Leishmania). The Friedman and the Mann Whitney tests were used for statistical analysis. The Friedman test was used to analyze each segment of the GIT within each group of dogs and the Mann Whitney test was used to compare the GIT segments between clinically unaffected and affected dogs.
The infected dogs had an increased number of macrophages, plasma cells and lymphocytes, but lesions were generally mild. Parasite distribution in the GIT was evident in all intestinal segments and layers of the intestinal wall (mucosal, muscular and submucosal) irrespective of the clinical status of the dogs. However, the parasite load was statistically higher in the caecum and colon than in other segments of the GIT.
The high parasite burden evident throughout the GIT mucosa with only mild pathological alterations led us to consider whether Leishmania gains an advantage from the intestinal immunoregulatory response (immunological tolerance).
PMCID: PMC3269393  PMID: 22166041
9.  Genetic evaluation of Addison's disease in the Portuguese Water Dog 
Addison's disease, also known as hypoadrenocorticism, has been reported in many individual dogs, although some breeds exhibit a greater incidence than the population as a whole. Addison's is presumed to be an autoimmune mediated hereditary defect but the mode of inheritance remains unclear. In particular, the heritability and mode of inheritance have not been defined for the Portuguese Water Dog although Addison's is known to be prevalent in the breed.
The analyses present clear evidence that establishes Addison's disease as an inherited disorder in the Portuguese Water Dog with an estimate of heritability of 0.49 (± 0.16); there were no differences in risk for disease across sexes (p > 0.49). Further, the complex segregation analysis provides suggestive evidence that Addison's disease in the Portuguese Water Dog is inherited under the control of a single, autosomal recessive locus.
The high heritability and mode of inheritance of Addison's disease in the Portuguese Water Dog should enable the detection of segregating markers in a genome-wide scan and the identification of a locus linked to Addison's. Though the confirmation of Addison's disease as an autosomal recessive disorder must wait until the gene is identified, breeders of these dogs may wish to keep the present findings in mind as they plan their breeding programs to select against producing affected dogs.
PMCID: PMC1481556  PMID: 16670022
10.  Bartonella vinsonii subsp. berkhoffii and Related Members of the Alpha Subdivision of the Proteobacteria in Dogs with Cardiac Arrhythmias, Endocarditis, or Myocarditis 
Journal of Clinical Microbiology  1999;37(11):3618-3626.
Cardiac arrhythmias, endocarditis, or myocarditis was identified in 12 dogs, of which 11 were seroreactive to Bartonella vinsonii subspecies berkhoffii antigens. Historical abnormalities were highly variable but frequently included substantial weight loss, syncope, collapse, or sudden death. Fever was an infrequently detected abnormality. Cardiac disease was diagnosed following an illness of short duration in most dogs, but a protracted illness of at least 6 months' duration was reported for four dogs. Valvular endocarditis was diagnosed echocardiographically or histologically in eight dogs, two of which also had moderate to severe multifocal myocarditis. Four dogs lacking definitive evidence of endocarditis were included because of seroreactivity to B. vinsonii antigens and uncharacterized heart murmurs and/or arrhythmias. Alpha proteobacteria were not isolated from the blood by either conventional or lysis centrifugation blood culture techniques. Using PCR amplification and DNA sequencing of a portion of the 16S rRNA gene, B. vinsonii was identified in the blood or heart valves of three dogs. DNA sequence alignment of PCR amplicons derived from blood or tissue samples from seven dogs clustered among members of the alpha subdivision of the Proteobacteria and suggested the possibility of involvement of one or more alpha proteobacteria; however, because of the limited quantity of sequence, the genus could not be identified. Serologic or molecular evidence of coinfection with tick-transmitted pathogens, including Ehrlichia canis, Babesia canis, Babesia gibsonii, or spotted fever group rickettsiae, was obtained for seven dogs. We conclude that B. vinsonii subsp. berkhoffii and closely related species of alpha proteobacteria are an important, previously unrecognized cause of arrhythmias, endocarditis, myocarditis, syncope, and sudden death in dogs.
PMCID: PMC85709  PMID: 10523564
11.  Evaluation of dental socket healing after using of porous titanium granules: Histologic and histomorphometric assessment in dogs 
Dental Research Journal  2012;9(5):600-606.
Different methods have been suggested to preserve bone architecture following traumatic events such as teeth extraction. The purpose of the study was to histologically and histomorphometrically evaluate the dental socket healing after applying porous titanium granules (PTG) in dogs.
Materials and Methods:
Four healthy male dogs were involved in the present 6-weeks experimental animal study. Three sockets were surgically created in each side of dog's mandible. One of the sockets in one side was randomly filled by PTG and covered by a resorbable membrane (Tigran + membrane group). Another socket was left unfilled and just covered by the same membrane (membrane group) and the last one was left unfilled and uncovered as the control group. The dogs were killed at two time intervals (2 weeks and 6 weeks, two dogs at each time point). All samples were histologically evaluated under an optical microscope for a new bone formation. Data were analyzed by SPSS ver. 16 and Kruskal–Wallis and Mann–Whitney tests were used to compare data in different groups (α = 0.05).
There was a significant difference between the Tigran + membrane and the control group in 2 and 6 weeks in the mean amount of total regenerated bone (P < 0.05). The mean amounts of woven, lamellar, and total regenerated bone showed significant differences between 2 weeks and 6 weeks for all three groups (P < 0.05).
It can be assumed that the use of Tigran bone substitute with membrane can promote the bone regeneration in bone defects.
PMCID: PMC3612198  PMID: 23559926
Bone healing; dental socket; membrane; porous titanium granules
12.  Borrelia burgdorferi migrates into joint capsules and causes an up-regulation of interleukin-8 in synovial membranes of dogs experimentally infected with ticks. 
Infection and Immunity  1997;65(4):1273-1285.
Twenty 6-week-old specific-pathogen-free beagles were infected with Borrelia burgdorferi by tick challenge, and five uninfected dogs served as controls. During the study, all dogs were monitored for infection, clinical signs, and antibody response against B. burgdorferi. During episodes of lameness or postmortem, synovial fluids from each dog were examined for volume, cell number, polymorphonuclear leukocyte (PMN) content, cell viability, and chemotactic activity. Twenty-five tissues collected postmortem from each dog were tested for interleukin-8 (IL-8) mRNA, tumor necrosis factor alpha (TNF-alpha) mRNA, presence of live spirochetes, and histopathological changes. Thirteen infected dogs (group A), which seroconverted rapidly (maximum titers within 50 to 90 days), developed acute and severe mono- or oligoarthritis almost exclusively in the limb closest to the tick bite (median incubation period, 66 days). Synovial fluids of the arthritic joints collected during episodes of lameness had significantly elevated volume, cell count, PMN proportion, cell viability, and chemotactic activity for PMNs. The remaining joints of the same animals contained synovial fluids with elevated chemotactic activity and cell viability. Twelve dogs tested positive for IL-8 mRNA in multiple tissues (synovia, pericardium, and peritoneum), and 10 dogs expressed TNF-alpha mRNA, but only in the tributary lymph nodes of the inflamed joints. Histological examinations revealed severe poly- or oligoarthritis and moderate to severe cortical hyperplasia in draining lymph nodes of the inflamed joints in all 13 dogs. Seven infected dogs with mild or no clinical signs (group B) seroconverted slowly (peak titers after 90 days), and only some joint fluids showed chemotactic activity, which on average was lower than that in inflamed and noninflamed joints from dogs in group A. Four dogs expressed IL-8 mRNA (in the synovia and pericardium), and three dogs had TNF-alpha mRNA in tributary lymph nodes. Histologically, nonsuppurative arthritis was found in multiple joints, and mild to moderate cortical hyperplasia was found in draining lymph nodes. Five uninfected dogs without lameness (group C) had normal synovial fluids and tissues. In all infected dogs, live spirochetes were demonstrated more frequently in tissues of the somatic quadrant closest to the tick bite than in tissues further from the site of infection, suggesting that dissemination of B. burgdorferi occurs more by migration than by blood-borne spread. From these studies employing a canine model of B. burgdorferi infection, we conclude that IL-8 is involved in the pathogenesis of acute Lyme arthritis.
PMCID: PMC175128  PMID: 9119462
13.  Failure of nebulized irritant, acidic, or hypotonic solutions or external mechanical stimulation of the trachea to consistently induce coughing in healthy, awake dogs 
A useful approach for evaluating antitussive drugs in humans is to determine the sensitivity of the cough reflex to a standard challenge. The purpose of this study was to determine if methods used to induce coughing in humans would be effective when used on awake, untrained, healthy dogs for future application in therapeutic trials involving dogs with spontaneous disease. Methods tested were: mechanically stimulating the trachea by digital compression as well as by vibration from an electric shaver, neck massager, and palm sander (11 dogs), and administering nebulized irritant (3000 μM capsaicin), acidic (1 M citric acid), and hypotonic (deionized water) solutions using face masks (4 dogs). The threshold for success was defined as induction of at least 2 moderate or strong coughs in at least 75% of the dogs. None of the methods tested was successful. Digital compression induced soft (n = 2) or moderate (n = 1) coughing in 3 of 11 dogs tested. Nebulization of citric acid induced 1 soft cough in 1 of 4 dogs. It was concluded that coughing cannot be successfully induced in awake, healthy dogs using methods that are successful in humans. Other strategies must be developed so that cough sensitivity can be objectively and non-invasively measured in dogs for clinical research purposes.
PMCID: PMC3122971  PMID: 22211000
Twelve control dogs receiving a single intravenous injection of mercuric chloride, 3.0 mg. per kg., all died within 4 to 11 days afterwards with marked nitrogen retention and extensive necrosis and calcification of the epithelium lining the proximal convoluted tubules. Three dogs of comparable age and weight were reduced to a standard hypoproteinemic state by repeated plasmapheresis. Each dog then received the same dose of mercuric chloride as the controls. None of these dogs became sick, none showed any elevation of non-protein nitrogen, and the kidneys— both in the gross and histologically—appeared normal when they were examined 8 to 45 days later. As tested thus far intensive plasmapheresis following the injection of mercuric chloride has been without effect in preventing the classical changes of mercuric chloride injury observed in the control dogs. The simplest explanation for these phenomena is that mercuric chloride acts on a more or less specific substance (presumably fabricated or concentrated in the renal cortex) which is depleted in the standard hypoproteinemic state. Other possibilities are mentioned. These findings are in sharp contrast to the results of similar experiments with uranium nitrate. The hypoproteinemic state appears to render the animals more susceptible to uranium injury (3). This probably indicates that the mode of action of the two heavy metals is different.
PMCID: PMC2135285  PMID: 19871254
15.  The effects of the Ilizarov distraction technique on bone and muscle in a canine model: a preliminary report. 
This study examined the functional and morphological changes experienced by bone and muscle during Ilizarov distraction osteogenesis. Although extensive research has been conducted in the area of regenerate bone formation, the effect of limb lengthening on the biomechanical properties of bone and muscle has not been thoroughly addressed. In this study, an Ilizarov external fixator was applied to one tibia of nine skeletally mature dogs, and distracted 3 cm over thirty days. The contralateral tibia served as control. Histology and weekly radiographs assessed muscle morphology and bone growth. The contractile capabilities of the gastrocnemius muscles from the experimental and control limbs were measured prior to sacrifice, and the bending stiffness of the tibias of five dogs was determined. All dogs experienced loss of knee extension secondary to muscle contracture and/or stiffness about the joint. These dogs did not bear weight on the experimental limb. In one dog, spontaneous resolution of the muscle contracture allowed partial weight bearing during the last three weeks of consolidation. Despite 3 cm distraction, tibial lengthening ranged from 1.7 to 3 cm. Biomechanical testing revealed a significant reduction in the bending stiffness of the lengthened bones when compared with control values (p < 0.003). The weight of the lengthened muscles was 35% less than control values, a finding consistent with the histology which showed mild muscle fiber degeneration in all dogs. The contractile capabilities of the lengthened muscles were reduced to 29-80% of control values (p < 0.005). In contrast, the lengthened muscle from the weight bearing dog retained 85% of the weight and 104% of the maximum contractile force of the control muscle.
PMCID: PMC2378139  PMID: 9129270
16.  An Unusual Case of Relay Pentobarbital Toxicosis in a Dog 
Journal of Medical Toxicology  2011;7(3):236-239.
Sodium pentobarbital and phenytoin are common constituents of veterinary euthanasia solutions in the United States. Relay, or secondary, barbiturate toxicosis has been reported in carnivorous animals that have fed from the carcasses of euthanized livestock. This case report presents barbiturate toxicosis in a dog. A 2-year-old female spayed Australian shepherd presented comatose 2 h after ingesting an unknown substance on the beach. The material was retrieved from the stomach by gastric lavage and visually identified as fish or other animal tissue. The dog recovered with symptomatic and supportive therapy and was released on the third day of hospitalization. Tissue found on the beach near where the dog walked and a urine sample from the dog were analyzed by gas chromatography/mass spectrometry. Both samples were positive for pentobarbital and phenytoin. The tissue was consistent with mammalian blubber based on gross and histological examination. Three weeks previously, a juvenile humpback whale had stranded on the beach where the dog had ingested the unknown substance. The whale had been euthanized with a barbiturate solution, necropsied, and removed from the beach. It was not definitively determined that the pentobarbital-containing blubber ingested by the dog was from the euthanized whale, but that was the most likely source. Although attempts were made to remove the whale’s remains from the beach, practical considerations made complete removal challenging, if not impossible.
PMCID: PMC3550208  PMID: 21660622
Barbiturates; Dog; Pentobarbital; Relay toxicosis; Secondary poisoning
17.  Effect of enamel matrix derivative on bone formation around intraosseous titanium implant: An experimental study in canine model 
Dental Research Journal  2012;9(6):790-796.
The purpose of this study was to perform a histological, histomorphometrical, and immunohistochemical evaluation of the effect of Enamel matrix derivative (EMD) on bone formation around titanium dental implant.
Materials and Methods:
In this animal study, 12 implants (10 × 3.8 mm) were inserted in the tibia bone of three dogs of Iranian breed. Two implants were placed in each tibia with EMD only on the left side. The dogs were sacrificed 2, 4, and 6 weeks after implantation. Following decalcification of the implants’ surrounding tissue and preparation of 4 μm thick sections, they were stained with hematoxylin and eosin (H and E) and immunohistochemical (IHC) stain for osteopontin (OPN) marker. Histomorphometric evaluation was performed via measurement of the percentage of the woven, lamellar, and total generated bone. Light microscopy osteoblastic intensity of OPN in osteoblasts and bone matrix was also evaluated Data were analyzed by Wilcoxon signed Ranks, and Mc Nemar tests.
In both control and EMD-applied groups, bone formation was recognized around the implants at the 4th week postimplantation. The percentage of total generated bone in the test group was higher than the control group, although being not statistically significant (P value = 0.917). Osteoclasts exhibited significantly higher proliferation activity compared the control group when stimulated by EMD (P value = 0.027). On average, the staining intensity in osteoblasts and extracellular matrix of bone, in EMD-applied subjects was higher than those of the controls (P value = 0.167 and P value = 0.414, respectively).
EMD enhanced bone formation around dental implants, but this increase was not significant.
PMCID: PMC3612232  PMID: 23559960
Bone formation; dental implant; enamel matrix protein; osteopontin
18.  Helicobacter pylori gastric infection in gnotobiotic beagle dogs. 
Infection and Immunity  1990;58(8):2606-2612.
Establishment of infection with Helicobacter pylori and gastritis in nonhuman species is currently only successful in gnotobiotic piglets. This study was designed to determine whether H. pylori will colonize the gastrointestinal tract of gnotobiotic dogs. Gnotobiotic beagle pups were derived by standard methods. Group A (five dogs) was orally challenged with 3 x 10(8) H. pylori at 7 days of age. Group B (two dogs) received only peptone water but was contact-exposed beginning on day 23 postinfection (p.i.). Necropsy was performed on dogs on day 30 p.i. H. pylori colonized the stomach of all dogs (groups A and B). Urease map analysis correlated with the microbiologic findings and indicated that the density of colonization was less than that observed in human tissue. Organisms were also recovered from the pharynx, esophagus, duodenum, and rectum of 1, 2, 2, and 1 dog, respectively. All group A and one group B dog developed serum immunoglobulin G specific for H. pylori by day 30 p.i. Gross lesions were restricted to the stomach and consisted of small (less than 1 mm) lymphoid follicles. Microscopically, there were focal to diffuse lymphoplasmacytic infiltrates with follicle formation and mild to moderate infiltration of neutrophils and eosinophils in the gastric lamina propria. With the Warthin-Starry silver stain, organisms were seen on the surface of the gastric epithelial cells, beneath the mucus layer. We conclude that H. pylori colonizes the stomachs of gnotobiotic dogs for at least 1 month and the lesions resemble those seen in humans. H. pylori is transmissible by contact from infected to noninfected dogs.
PMCID: PMC258862  PMID: 2370111
19.  Genetic regulation of canine skeletal traits: trade-offs between the hind limbs and forelimbs in the fox and dog 
Genetic variation in functionally integrated skeletal traits can be maintained over 10 million years despite bottlenecks and stringent selection. Here, we describe an analysis of the genetic architecture of the canid axial skeleton using populations of the Portuguese Water Dog Canis familiaris) and silver fox (Vulpes vulpes). Twenty-one skeletal metrics taken from radiographs of the forelimbs and hind limbs of the fox and dog were used to construct separate anatomical principal component (PC) matrices of the two species. In both species, 15 of the 21 PCs exhibited significant heritability, ranging from 25% to 70%. The second PC, in both species, represents a trade-off in which limb-bone width is inversely correlated with limb-bone length. PC2 accounts for approximately 15% of the observed skeletal variation, ~30% of the variation in shape. Many of the other significant PCs affect very small amounts of variation (e.g., 0.2–2%) along trade-off axes that partition function between the forelimbs and hind limbs. These PCs represent shape axes in which an increase in size of an element of the forelimb is associated with a decrease in size of an element of the hind limb and vice versa. In most cases, these trade-offs are heritable in both species and genetic loci have been identified in the Portuguese Water Dog for many of these. These PCs, present in both the dog and the fox, include ones that affect lengths of the forelimb versus the hind limb, length of the forefoot versus that of the hind foot, muscle moment (i.e., lever) arms of the forelimb versus hind limb, and cortical thickness of the bones of the forelimb versus hind limb. These inverse relationships suggest that genetic regulation of the axial skeleton results, in part, from the action of genes that influence suites of functionally integrated traits. Their presence in both dogs and foxes suggests that the genes controlling the regulation of these PCs of the forelimb versus hind limb may be found in other tetrapod taxa.
PMCID: PMC2367254  PMID: 18458753
20.  Bilaterally Asymmetric Effects of Quantitative Trait Loci (QTLs): QTLs That Affect Laxity in the Right Versus Left Coxofemoral (Hip) Joints of the Dog (Canis familiaris) 
In dogs hip joint laxity that can lead to degenerative joint disease (DJD) is frequent and heritable, providing a genetic model for some aspects of the human disease. We have used Portuguese water dogs (PWDs) to identify Quantitative trait loci (QTLs) that regulate laxity in the hip joint.A population of 286 PWDs, each characterized by ca. 500 molecular genetic markers, was analyzed for subluxation of the hip joint as measured by the Norberg angle, a quantitative radiographic measure of laxity. A significant directed asymmetry was observed, such that greater laxity was observed in the left than the right hip. This asymmetry was not heritable. However, the average Norberg angle was highly heritable as were the Norberg angles of either the right or left hips. After correction for pedigree effects, two QTLs were identified using the metrics of the left and right hips as separate data sets. Both are on canine chromosome 1 (CFA1), separated by about 95 Mb. One QTL, associated with the SSR marker FH2524 was significant for the left, but not the right hip. The other, associated with FH2598, was significant for the right but not the left hip. For both QTLs, some extreme phenotypes were best explained by specific interactions between haplotypes.
PMCID: PMC2778498  PMID: 14708095
quantitative trait loci (QTLs); dog; hip laxity; bilateral asymmetry; Norberg angle; Canine genetics; hip dysplasia
21.  Histopathological analysis of corticosteroid-antibiotic preparation and propolis paste formulation as intracanal medication after pulpectomy: an in vivo study 
Intracanal medication in pulpectomy therapy is used between appointments with the objective of reducing pain and inflammatory processes in pulp and periapical tissues. Propolis has been known as a natural antibiotic and has been subject of medical and dental research due to its therapeutic properties such as antibiotic, analgesic and anti-inflammatory effects.
The aim was to carry out an in vivo evaluation of the periapical tissue response to propolis paste when used as an intracanal medication in the teeth of dogs after pulpectomy.
Material and Methods
72 dog's incisors were selected for the experiment. After biomechanical preparation the root canal was filled with a corticosteroid-antibiotic preparation, experimental propolis paste, non-medicament (negative control) or non-pulpectomy at all (positive control). The medications were left inside the root canal for 7, 14 or 28 days. At the end of the experimental period histological sections were prepared and all laboratories processes for Harris hematoxylin and eosin staining was proceeded followed by the analysis using an optical microscope. Sections were classified according to a score representing the inflammatory events observed: the presence of polymorphonuclear neutrophils, polymorphonuclear eosinophils, lymphocytes and plasma cells, macrophages and/or giant cells, fibrous condensation and abscesses.
There were statistically significant differences between the tissue reactions caused by the two substances being tested, after different experimental periods, with the periapical tissue that was in contact with propolis paste exhibiting fewer inflammatory reactions in comparison to corticosteroid-antibiotic preparation.
The low tissue responses from propolis paste suggest that this material could be considered as an option for root canal medication after pulpectomy.
PMCID: PMC3928772  PMID: 22437678
Endodontics; Propolis; Pulpectomy; Dental pulp cavity
22.  Cliniconeuropathologic findings of familial frontal lobe epilepsy in Shetland sheepdogs 
We examined an epileptic focus by electroencephalography (EEG) by using an international 10-20 electrode system in 11 Shetland sheep dogs affected with familial idiopathic epilepsy. We also performed an evaluation of the amino acids in the cerebrospinal fluid (CSF) and a pathologic examination of the brains of 8 dogs that died from status epilepticus. Continuous electroencephalography demonstrated that an epileptic focus was initially detected in the frontal lobe, particularly the internal area, and that paroxysmal foci developed diffusely in other lobes of affected dogs with recurrent convulsions. The EEG analyses indicated spike and sharp wave complexes, which were considered to be paroxysmal discharges. An increased value for glutamate or aspartate was found in the CSF of some epileptic dogs. Histologically, acute neuronal necrosis and astrocytosis were distributed predominantly in the cingulate cortex and internal area of frontal cortex, less frequently in other areas of the cerebrum. The results of this study suggest that, initially, the dogs have an epileptic focus in the frontal lobe, and that the focus extends gradually to other areas of the cerebrum. Based on the distribution of neuronal necrosis and astrocytosis, acute neuronal damage may be related to the superexcitation of neurons following epilepsy.
PMCID: PMC226980  PMID: 11858647
23.  Hepatocellular carcinoma in a young dog 
The Canadian Veterinary Journal  2013;54(9):845-848.
A 25-month-old Chihuahua dog with no clinical signs was evaluated for high serum liver enzymes. Ultrasonography and computed tomography revealed a mass in the left hepatic medial lobe. The histological diagnosis reached using resected tissues was hepatocellular carcinoma (HCC). To the authors’ knowledge, this is the youngest dog diagnosed with HCC.
PMCID: PMC3743567  PMID: 24155487
24.  Pathogenesis and phylogenetic analyses of canine distemper virus strain ZJ7 isolate from domestic dogs in China 
Virology Journal  2011;8:520.
A new isolate of canine distemper virus (CDV), named ZJ7, was isolated from lung tissues of a dog suspected with CDV infection using MDCK cells. The ZJ7 isolate induced cytopathogenic effects of syncytia in MDCK cell after six passages. In order to evaluate pathogenesis of ZJ7 strain, three CDV sero-negative dogs were intranasally inoculated with its virus suspension. All infected dogs developed clinical signs of severe bloody diarrhea, conjunctivitis, ocular discharge, nasal discharge and coughing, fever and weight loss at 21 dpi, whereas the mock group infected with DMEM were normal. The results demonstrated that CDV-ZJ7 strain isolated by MDCK cell was virulent, and the nucleotide and amino acid sequences of strain ZJ7 had no change after isolation by MDCK cell when compared with the original virus from the fresh tissues. Molecular and phylogenetic analyses for the nucleocapsid (N), phosphoprotein (P) and receptor binding haemagglutinin (H) gene of the ZJ7 isolate clearly showed it is joins to the Asia 1 group cluster of CDV strains, the predominant genotype in China.
PMCID: PMC3229531  PMID: 22087872
Canine distemper virus (CDV); MDCK; Genotype; Phylogenetic analysis; Pathogenesis; Virulence
25.  A blinded randomised controlled trial to determine the effect of enteric coating on enzyme treatment for canine exocrine pancreatic efficiency 
Enzyme treatment is the mainstay for management of exocrine pancreatic insufficiency (EPI) in dogs. ‘Enteric-coated’ preparations have been developed to protect the enzyme from degradation in the stomach, but their efficacy has not been critically evaluated. The hypothesis of the current study was that enteric coating would have no effect on the efficacy of pancreatic enzyme treatment for dogs with EPI.
Thirty-eight client-owned dogs with naturally occurring EPI were included in this multicentre, blinded, randomised controlled trial. Dogs received either an enteric-coated enzyme preparation (test treatment) or an identical preparation without the enteric coating (control treatment) over a period of 56 days.
There were no significant differences in either signalment or cobalamin status (where cobalamin deficient or not) between the dogs on the test and control treatments. Body weight and body condition score increased in both groups during the trial (P<0.001) but the magnitude of increase was greater for the test treatment compared with the control treatment (P<0.001). By day 56, mean body weight increase was 17% (95% confidence interval 11-23%) in the test treatment group and 9% (95% confidence interval 4-15%) in the control treatment group. The dose of enzyme required increased over time (P<0.001) but there was no significant difference between treatments at any time point (P=0.225). Clinical disease severity score decreased over time for both groups (P=0.011) and no difference was noted between groups (P=0.869). No significant adverse effects were reported, for either treatment, for the duration of the trial.
Enteric coating a pancreatic enzyme treatment improves response in canine EPI.
PMCID: PMC3412697  PMID: 22839732
Dog; Pancreas; Malabsorption; Diarrhoea; Lipase; Trypsin

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