A pathological study of the salivary glands of rabid dogs in the Philippines

Rabies is a zoonotic disease caused by the rabies virus. While the salivary glands are important as exit and propagation sites for the rabies virus, the mechanisms of rabies excretion remain unclear. Here, we investigated the histopathology of the salivary glands of rabid dogs and analyzed the mechanism of excretion into the oral cavity. Mandibular and parotid glands of 22 rabid dogs and three control dogs were used. Mild to moderate non-suppurative sialadenitis was observed in the mandibular glands of 19 of the 22 dogs, characterized by loss of acinar epithelium and infiltration by lymphoplasmacytic cells. Viral antigens were detected in the mucous acinar epithelium, ganglion neurons and myoepithelium. Acinar epithelium and lymphocytes were positive for anti-caspase-3 antibodies and TUNEL staining. In contrast, no notable findings were observed in the ductal epithelial cells and serous demilune. In the parotid gland, the acinar cells, myoepithelium and ductal epithelium all tested negative. These findings confirmed the path through which the rabies virus descends along the facial nerve after proliferation in the brain to reach the ganglion neurons of the mandibular gland, subsequently traveling to the acinar epithelium via the salivary gland myoepithelium. Furthermore, the observation that nerve endings passing through the myoepithelium were absent from the ductal system suggested that viral proliferation and cytotoxicity could not occur there, ensuring that secretions containing the virus are efficiently excreted into the oral cavity.     

Salivary gland necrosis in dogs: a retrospective study of 19 cases

Salivary gland necrosis has been described in dogs and is characterised by enlarged, hard, painful salivary glands, retching and vomition or regurgitation. The cause has yet to be determined. A retrospective study of 19 dogs with the same clinical signs was undertaken for breed, age, gender, history and presenting signs, diagnostic evaluations and findings, treatment and outcome. An underlying association was identified in 16 of the 19 dogs. This included Spirocerca lupi infestation (seven dogs), megaoesophagus (three dogs) and oesophagitis, oesophageal diverticulum, giardiasis and suspected autoimmune sialadenitis. Almost all associated lesions involved the oesophagus. Where the associated disease was successfully treated, the salivary glands returned to normal and all clinical signs resolved. It is hypothesised that an afferent vagal reflex may be involved, and that the mechanism of disease is similar to the neural pathogenesis suggested for hypertrophic osteopathy; in this instance, the efferent targets are the salivary glands rather than the limbs.     

Idiopathic phenobarbital‐responsive hypersialosis in the dog: an unusual form of limbic epilepsy?

Three unusual cases of salivary gland enlargement and hypersialosis in the dog that responded to anticonvulsant therapy are reported. Presenting complaints included weight loss, hypersalivation, retching and vomiting of several weeks' duration. Two dogs were presented with enlarged painful mandibular salivary glands. The third dog exhibited bizarre behaviour (including jaw chattering) and developed enlarged painful mandibular salivary glands during hospitalisation. Fine needle aspirate cytology and biopsies from the enlarged salivary glands revealed no significant pathological changes. In one dog, an electroencephalogram revealed changes consistent with epilepsy. Hypersialism and salivary gland enlargement resolved completely during phenobarbital administration in all cases. Two dogs were successfully weaned off treatment six months after diagnosis. The remaining dog relapsed after eight months, but normalised with the addition of oral potassium bromide. It is hypothesised that the syndrome idiopathic hypersialosis may in fact be an unusual form of limbic epilepsy.     

Phenobarbitone-responsive hypersialism in two dogs

Two dogs were examined for hypersialism associated with non-painful, symmetrical enlargement of both mandibular salivary glands. These signs were accompanied by inappetence and weight loss. The underlying cause of the problem could not be determined despite extensive investigations including fine needle aspiration biopsy of affected salivary glands. Hypersialism and mandibular salivary gland enlargement resolved completely during pheno-barbitone administration.     

Quantitation of histochemical staining of salivary gland mucin using image analysis in cats and dogs

Two different, computer-based, image analysis methods were employed to complement subjective assessment of patterns of mucin staining (by periodic acid Schiff/alcian blue, aldehyde fuchsin/alcian blue and potassium hydroxide-alcian blue/phenylhydrazine hydrocholoride) in digitised images of sections of major and minor salivary glands from cats and dogs. Image analysis based on red, green and blue (RGB) staining was not suitable for quantitation of histochemical staining of mucin in salivary glands. Image analysis based on hue, saturation and lightness (HSL) allowed quantitative assessment of staining by different stain components and of mixed staining, and enabled comparison of staining of different glands in dogs and cats. Quantitative analysis based on HSL allowed differentiation of differences in staining patterns of major and minor cat and dog salivary glands, and between the species; such differences would have been impossible to distinguish on subjective grounds alone. Quantitative assessment of normal salivary gland histochemistry allows comparison with staining patterns in disease.     

Immunohistochemical localization of CB1 receptor in canine salivary glands

CB1 is a member of the G-protein-linked receptor superfamily that is present in the central nervous system as well as in certain peripheral neuronal and non-neuronal tissues. Recently, the presence of CB1 was found in the ductal system of the major salivary glands of laboratory animals, but no data are available for domestic mammals. Thus, in the present study, we examined the presence and distribution of CB1 in the major salivary glands of dogs using immunohistochemical techniques. CB1 was found in the parotid and mandibular glands of adult dogs; positive immunoreaction was localized to the cells of the striated ducts, with a peculiar localization on or near the apical membrane. This particular localization may be explained by the characteristics of this receptor as membrane-associated. The acinar structures were completely negative for CB1. We conclude that CB1 is involved in the control of dog salivary secretion via endogenous substances, likely endocannabinoids. The localization of CB1 highlights that endocannabinoids promote qualitative and/or quantitative changes of the primary saliva in the ductal system.     

Pharyngeal mucoceles in dogs: 14 cases

This report describes the clinical features of a series of dogs with pharyngeal salivary mucoceles. A retrospective study of 14 dogs with pharyngeal mucocele was performed. Medical records from 1983 to 2003 were reviewed for information regarding signalment, clinical signs, diagnosis, surgical procedures, and short-term and long-term outcome. Miniature and toy poodles were common breeds in the study population, and 79% of the dogs were male. The most common presenting sign was dyspnea (50%). Diagnosis was by fine-needle aspirate, which revealed a mucoid substance in 93% of dogs. Histopathology of the excised salivary glands revealed lymphoplasmacytic inflammation in all dogs that had histopathology performed. Forty-three percent of the dogs had a cervical mucocele on the same side as the pharyngeal mucocele. Surgical therapy was performed in 13 dogs, which consisted of excision of the mandibular and sublingual salivary glands, excision of the mucocele, or marsupulization of the mucocele. Only two dogs had recurrence of the pharyngeal mucocele. In this study, pharyngeal mucoceles occurred in predominantly small dogs that frequently presented with respiratory signs. Surgical treatment was successful in most dogs.     

Histology of salivary gland infarction in the dog.

Salivary gland infarction was found in two adult dogs. The main changes were ischaemic necrosis, capsular fibrosis and regenerative hyperplasia of surviving ductal epithelium. Necrosis of arterial tunica media and thrombosis were found only in the infarcted parts of the salivary glands. The lesions appeared to be confined to the salivary glands; no cause of the infarction was found.     

CT sialography in the dog - a cadaver study

To document computed tomographic topography of salivary glands and their ducts in dogs, a retrograde filling with methylcellulose and iodinated contrast medium was performed in three cadavers. Demarcation of the parotid, mandibular and zygomatic glands was achieved. Surrounding structures were imaged without beam hardening artefacts. Landmarks for the parotid, mandibular and zygomatic glands were the external acoustic canal, the mandibular angle and the pterygopalatine fossa, respectively. Sialograms of the parotid, mandibular and zygomatic ducts were achieved, whereas neither the sublingual glands nor their ducts could be contrasted. The images provide a three-dimensional visualization of the salivary glands and their ducts.     

Pharyngeal Mucoceles in Dogs

Eight cases of pharyngeal mucoceles were diagnosed in 49 dogs with salivary mucoceles over a 7 year period. Five of the eight pharyngeal mucoceles were seen in Miniature Poodles. The presenting complaint in all eight dogs was labored breathing, with or without coughing upon excitement or exercise. Diagnosis was made by aspirating a thick mucoid fluid from the pharyngeal swelling. Treatment consisted of resection of the mandibular and sublingual salivary glands on the affected side and aspiration of the mucocele in all eight dogs, and resection of redundant pharyngeal tissue in five dogs. Time to follow-up ranged from 1.5 to 7.5 years. There were no postsurgical complications or recurrences of the mucocele.     

First case of salivary mucocele originating from the minor salivary gland of the soft palate in a dog

We found a case of salivary mucocele that originated in the minor salivary gland (palatine gland) of the soft palate in a dog. At first admission, the soft palate swelled remarkably. Computed tomography (CT) revealed cystic radiolucency inside a large quantity of liquid in the soft palate, and most of the airway was occupied. Marsupialization was performed, but since a recurrence was observed one month later, the salivary mucocele was removed. There has been no report of salivary mucocele arising from the minor salivary gland of the soft palate in dogs. To our knowledge, this case is the first. Complete removal, including minor salivary glands surrounding the lesion, is necessary for treatment of salivary mucocele in dogs.     

Keratoconjunctivitis sicca: immunological evaluation of 62 canine cases

The etiology of keratoconjunctivitis sicca (KCS) in 62 dogs was evaluated, using immunologic techniques. Using direct fluorescent antibody testing, autoantibodies within the lacrimal, salivary, or pancreatic glands were detected in 5 of 8 dogs tested. Circulating antibodies to the nictitating membrane gland, main lacrimal gland, parotid salivary gland, or mandibular salivary gland were detected using indirect fluorescent antibodies in 9 of 31, 3 of 31, 5 of 31, and 5 of 31 sera, respectively. Using radial immunodiffusion, hyper-gamma-globulinemia was detected in 21 of 30 dogs with KCS. Antinuclear antibodies, primarily in a nucleolar pattern, were demonstrated in 20 of 50 dogs with KCS. Lymphocytic infiltrates were evident in 5 of 9 labial salivary biopsies, 2 of 4 parotid gland specimens, 2 of 4 mandibular gland specimens, and 2 of 3 thyroid gland specimens taken from dogs with KCS. Autoimmune diseases had been previously documented in 4 of 62 dogs. Twenty-five of the 62 dogs (40%) had concurrent problems indicative of an underlying immunologic disorder.     

Adsorption of Wild Rabies Virus and Neurotropic-Adapted Viruses by Activated Attapulgite

The adsorption by attapulgite (a naturally occurring clay) of wild rabies virus in naturally rabid dogs was investigated. Attapulgite selectively absorbed rabies virus in the brain tissue of naturally rabid dogs and rejected rabies virus in extracts from the submaxillary salivary glands of the rabid dogs. Attapulgite rejected the neurotropic-adapted viruses lymphocytic choriomeningitis, Eastern equine and Western equine encephalitis, Japanese B encephalitis, and St. Louis encephalitis in the brain tissue of suckling mice.     

Immunofluorescence Study of Wild Rabies Virus and Antibody

A concentrate of wild rabies antibody was prepared from hyperimmune serums of three dogs refractory to wild rabies. The animals resisted repeated intramuscular injections of large doses of wild rabies virus in emulsions of whole brain, in emulsions of submaxillary salivary glands, and in emulsified mixtures of brain and submaxillary glands taken from naturally rabid dogs.

The antibody was conjugated with fluorochrome and then absorbed by a procedure that gave “cell-free” working solutions of fluorescent antibody. The procedure entailed parallel absorption steps with minced pathological canine submaxillary glands from (1) naturally rabid dogs (these glands contained specific, undegraded, natural antigens of live wild rabies virus plus nonspecific substances and antigens) and (2) nonrabid dogs from a rabies endemic region (these glands contained nonspecific substances and antigens).

Extracts from submaxillary glands of the three naturally rabid dogs and one nonrabid dog were stained with a cell-free solution of the fluorescent antibody. The glands of the rabid dogs contained fluorescent aggregates of intense green spherical and filamentous particles. When nonfluorescent canine hyperimmune serum was incubated with rabies-containing submaxillary extract, the rabies antigens were quenched. When nonfluorescent equine fixed virus antiserum was incubated with such extracts, the aggregates still retained bright fluorescence.

     

MR anatomy of salivary glands in the dog

This retrospective analysis documented the magnetic resonance imaging (MRI) appearance of normal salivary glands based on 101 studies in dogs with no detectable disease in the splanchnocranium. Surface, signal intensity, homogeneity, structure, symmetry and the relationship of glands to surrounding tissues were noted, and gland topography was assessed with E12 plastinated embedded sections. Signal intensity of salivary glands was isointense (7-40%) to hyperintense (60-90%) to muscle tissue on T1- and hyperintense on T2-weighted images. Salivary glands had an increased T1 signal after contrast medium was applied. Salivary gland structure appeared homogeneous in mandibular and major sublingual glands and heterogeneous in zygomatic and parotid glands. Consistent landmarks were the external auditory canal for parotid glands, the digastric muscle for mandibular and major sublingual glands, and the pterygopalatine fossa for zygomatic glands. The minor sublingual and ventral buccal glands could not be localized with low-field MRI.     

Electrophoresis Study of Extracts of Submaxillary Salivary Glands from Naturally Rabid Dogs

Rabies virus in submaxillary salivary glands from naturally infected dogs was investigated by a paper electrophoresis technique, and the virus activity was quantitated by intracerebral titration in mice. Extracts from these salivary glands were found to contain (a) 10⁴ to 10⁶ mouse ICLD ₅₀ units of wild rabies seed and (b) a protein complex that migrated electrophoretically in the albumin band and more conspicuously in the beta and gamma bands of normal horse serum. The protein complex was interpreted to comprise aggregates of neutralizing antirabies antibody.     

Idiopathic salivary gland enlargement (sialadenosis) in dogs: a microscopic study

A histological, histochemical and morphometric study was performed on submandibular salivary glands from 13 dogs which had presented with a submandibular mass or swelling that proved to be a portion of non-inflammatory and non-neoplastic submandibular salivary gland. There were no consistent changes in lectin-binding histochemistry or immunohistochemical expression of various cell markers, and, in most cases, there was no measurable difference in acinar size in the affected gland. The possible explanation for the clinical salivary gland enlargement is therefore unclear.     

Safety evaluation of the SAG2 rabies virus mutant in Tunisian dogs and several non-target species.

The safety of the SAG2 rabies virus, a highly attenuated mutant of the SAD strain intended to vaccinate dogs by the oral route, was evaluated in local Tunisian dogs and in five other local species likely to consume vaccine baits. These species were the domestic cat (Felis catus), the jackal (Canis aureus), the jerboa (Jaculus orientalis), the merion (Meriones sp.) and the gerbil (Gerbillus campestris). The vaccine was administered orally to 21 dogs, 11 cats and eight jackals and orally or intramuscularly to 62 wild rodents of the above-mentioned species. Seven dogs, one cat, five jackals all juvenile and with poor health status) and two rodents died for intercurrent causes. The others were observed for 60-180 days. No animal showed any rabies symptom. Seroneutralizing antibodies were observed in all experimental groups, only after vaccination, with the highest rate being observed in jackals and rodents. The rabies virus was detected in the oral cavity of three cats 6 h after oral instillation, but was not isolated later either in saliva or in salivary glands. Tissue samples (brain and salivary glands) from dead or euthanized animals were examined for the rabies virus antigen by a fluorescent antibody test. No rabies antigen was detected. These trials confirm the safety of the SAG2 strain on the Tunisian species already demonstrated by other authors on many other target and non target species.     

Putative salivary allergens of the cat flea, Ctenocephalides felis felis.

The cat flea, Ctenocephalides felis felis, is the major initiator of flea bite hypersensitivity in dogs. Previous analyses of whole extracts of the flea and flea salivary secretions have failed to identify the allergens responsible. We dissected >2000 salivary glands from adult female fleas, extracted them into buffered saline containing protease inhibitors and fractionated the extract using gel permeation HPLC. Dogs were classified as hypersensitive to fleas (flea-feeding positive, FF+) or insensitive (flea-feeding negative, FF-) using a provocative test with live fleas. The allergenicity of the components of the salivary gland extract was tested by intradermal injection of samples of the column eluates. Dogs were also injected intradermally with a sample of whole salivary gland extract, and with histamine as a positive control. Negative control injections consisted of eluate from the column collected prior to fractions containing any protein. The skin of FF- dogs either did not respond or had a minimal response (a bleb approximately 2 mm larger than the injection blebs at the negative control injection sites) to all fractions and to the whole extract; histamine control injections produced positive responses (defined as wheals 5 mm greater than the blebs at the negative control injection sites) in all dogs. The skin of three of the nine FF+ dogs reacted positively to injection of a fraction containing protein/s with apparent MW 40k. Five other FF+ dogs reacted positively to the fractions containing proteins with apparent MW 12-8k. A single dog responded with very large, red wheals to injection of both the approximately MW 40k and MW12-8k fractions. These findings suggest that proteins with apparent MW 40k and MW 12k-8k are important in flea bite hypersensitivity. This work also supports a previous finding that mice which had been exposed to flea bites had antibodies to proteins with approximately MW 40k that were detected in salivary secretions of the flea.