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The pathogenesis of anal sacculitis has not been extensively investigated, although atopic dogs seem to be predisposed to the disease. The aim of this study was therefore to characterize and compare the bacterial microbiota and pro-inflammatory cytokines in the anal sacs of dogs from three groups (healthy dogs, untreated atopic dogs and atopic dogs receiving antipruritic treatment or allergen-specific immunotherapy) in order to determine whether changes could be at the origin of anal sacculitis in atopic dogs. Bacterial populations of anal sac secretions from fifteen healthy dogs, fourteen untreated and six treated atopic dogs were characterized by sequencing the V4 region of the 16S rRNA gene using Illumina technology. Proinflammatory cytokines were analyzed with the Luminex multiplex test. Community membership and structure were significantly different between the anal sacs of healthy and untreated atopic dogs (P = 0.002 and P = 0.003, respectively) and between those of untreated and treated atopic dogs (P = 0.012 and P = 0.017, respectively). However, the community structure was similar in healthy and treated atopic dogs (P = 0.332). Among the proinflammatory cytokines assessed, there was no significant difference between groups, except for interleukin 8 which was higher in the anal sacs of untreated atopic dogs compared to treated atopic dogs (P = 0.02), and tumor necrosis factor-alpha which was lower in the anal sacs of healthy dogs compared to treated atopic dogs (P = 0.04). These results reveal a dysbiosis in the anal sacs of atopic dogs, which may partially explain the predisposition of atopic dogs to develop bacterial anal sacculitis. Treatments received by atopic dogs (oclacitinib, desloratadine and allergen-specific immunotherapy) shift the microbiota of the anal sacs towards that of healthy dogs. Further studies are required to identify significant cytokines contributing to anal sacculitis in atopic dogs.

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The aim of the present study was to characterize the bacterial microbiota of anal sacs in healthy dogs using NGS. Swabs were used to sample the rectum and secretions from each anal sac in 15 healthy dogs. DNA was extracted from swabs and the V4 hypervariable region of the 16S rRNA gene was amplified and sequenced with Illumina MiSeq. Overall, 14 different bacterial phyla were identified in the rectum and in both anal sacs, the 5 main ones being Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, and Fusobacteria. The rectum had higher microbial diversity and richness than the left and right anal sacs. Community membership and structure significantly differed between the rectum and both anal sacs, but not between the right and the left anal sacs. This study showed that the diversity and richness of the bacterial microbiota of the anal sacs in dogs is greater than what has been reported in previous studies with culture-based methods. In conclusion, the bacterial microbiota of the anal sacs in dogs varies between individuals and differs from the rectal bacterial microbiota.

L'objectif de la présente étude était de caractériser le microbiote bactérien des sacs anaux de chiens en santé en utilisant le séquençage d'ADN à haut débit. Des écouvillons ont été utilisés pour échantillonner le rectum et les sécrétions provenant de chaque sac anal chez 15 chiens en santé. L'ADN a été extrait des écouvillons et la région hypervariable V4 du gène codant pour l'ARN ribosomique 16S a été amplifiée et séquencée avec Illumina MiSeq. En tout, 14 phyla bactériens différents ont été identifiés dans les sacs anaux droit et gauche et le rectum, les cinq principaux étant Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria et Fusobacteria. Le microbiote bactérien du rectum avait une diversité et une richesse plus importantes que le microbiote bactérien des deux sacs anaux. L'appartenance à la communauté et sa structure différaient significativement entre le rectum et les sacs anaux, mais pas entre le sac anal droit et le sac anal gauche. Cette étude a démontré que la diversité et la richesse du microbiote bactérien des sacs anaux chez le chien sont plus importantes que ce qui a été rapportées dans les études précédentes avec des cultures. En conclusion, le microbiote bactérien des sacs anaux chez le chien varie d'un individu à l'autre et diffère de celui du rectum.(Traduit par les auteurs).

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In social animals, scent secretions and marking behaviors play critical roles in communication, including intraspecific signals, such as identifying individuals and group membership, as well as interspecific signaling. Anal sacs are an important odor producing organ found across the carnivorans (species in the mammalian Order Carnivora). Secretions from the anal sac may be used as chemical signals by animals for behaviors ranging from defense to species recognition to signaling reproductive status. In addition, a recent study suggests that domestic cats utilize short-chain free fatty acids in anal sac secretions for individual recognition. The fermentation hypothesis is the idea that symbiotic microorganisms living in association with animals contribute to odor profiles used in chemical communication and that variation in these chemical signals reflects variation in the microbial community. Here we examine the fermentation hypothesis by characterizing volatile organic compounds (VOC) and bacteria isolated from anal sac secretions collected from a male Bengal cat (Felis catus × Prionailurus bengalensis), a cross between the domestic cat and the leopard cat. Both left and right anal sacs of a male Bengal cat were manually expressed (emptied) and collected. Half of the material was used to culture bacteria or to extract bacterial DNA and the other half was used for VOC analysis. DNA was extracted from the anal sac secretions and used for a 16S rRNA gene PCR amplification and sequencing based characterization of the microbial community. Additionally, some of the material was plated out in order to isolate bacterial colonies. Three taxa (Bacteroides fragilis, Tessaracoccus, and Finegoldia magna) were relatively abundant in the 16S rRNA gene sequence data and also isolated by culturing. Using Solid Phase Microextraction (SPME) gas chromatography-mass spectrometry (GC-MS), we tentatively identified 52 compounds from the Bengal cat anal sac secretions and 67 compounds from cultures of the three bacterial isolates chosen for further analysis. Among 67 compounds tentatively identified from bacterial isolates, 51 were also found in the anal sac secretion. We show that the bacterial community in the anal sac consists primarily of only a few abundant taxa and that isolates of these taxa produce numerous volatiles that are found in the combined anal sac volatile profile. Several of these volatiles are found in anal sac secretions from other carnivorans, and are also associated with known bacterial biosynthesis pathways. This is consistent with the fermentation hypothesis and the possibility that the anal sac is maintained at least in part to house bacteria that produce volatiles for the host.

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The present study gives a detailed phenotypic and genotypic characterization of three Trueperella abortisuis strains isolated from a ten year old male Hovawart dog with an abscess of anal sac, from urine of an eight year old European shorthair cat with urolithiasis and nephrolithiasis and from a 14year old Maine Coon cat with a perianal abscess, respectively. All three strains could be identified phenotypically, by MALDI-TOF MS analysis and genotypically by sequencing the 16S rDNA and the molecular target genes gap and tuf. The present study gives a first description of T. abortisuis of this origin.

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We know that microbes contribute to the production of odors that some animals use to communicate, but how common is this phenomenon? Recent studies capitalizing on new molecular technologies are uncovering fascinating associations between microbes and odors of wild animals, but causality is difficult to ascertain. Fundamental questions about the nature of these unique host-microbe interactions also remain unanswered. For instance, do microbes benefit from signaling associations with hosts? How does microbial community structure influence signal production? How do hosts regulate microbes in order to generate appropriate signals? Here, we review the current state of knowledge on microbially produced signals in animals and discuss key research foci that can advance our understanding of microbial-based signaling in the animal world.

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This study was conducted to characterize the gross and cytological characteristics of secretions from normal feline anal sacs. Thirty cats with no recent history of anal-sac disease were selected consecutively and their anal sacs manually expressed. The color, cell counts, or presence of solid portions of the secretions in the youngest cats (<1 year old) were not significantly different from the rest. Young cats (<1 year old) tended to more often have watery secretions. The secretions contained 13 (median) epithelial cells per 600x magnification microscopic field, basophilic background debris, and a mixture of Gram-positive cocci (83 per field; median), Gram-negative cocci (38 per field; median), and Gram-negative rods (1.8 per field; median). Most secretions had occasional neutrophils, and a minority of cats had occasional yeasts in their secretion. Erythrocytes were found only rarely. Extreme heterogeneity existed in gross color, consistency, and amount of solid material. Hence, these latter parameters would not be valid indicators of anal-sac disease.

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Two experiments were designed to determine the efficacy of intraglandular antibiotic treatment in beef bulls. Experiment 1 was designed to evaluate the glandular tissue reaction to intraglandular antibiotic treatment. Experiment 2 was conducted to determine the efficacy intraglandular injection of antibiotics for the treatment of naturally occurring cases of vesicular adenitis. Healthy beef bulls (n=15), 2 and 3 years of age, were randomly allocated to three equal treatment groups to receive 10% of the daily recommended parenteral dose of penicillin, ceftiofur, or oxytetracycline in a volume of 6 mL injected directly into one of the vesicular glands. Ultrasonography was performed before, immediately after, and at 24, 48 and 168 h after intraglandular injection. The size and hardness of vesicular glands injected with oxytetracycline was greater (P<0.01) than those injected with ceftiofur. Ultrasonographic pixel intensity increased (P<0.01) after treatment with antibiotics, especially after treatment with oxytetracycline or penicillin. In Experiment 2, yearling beef bulls with clinical vesicular adenitis (n=14) were referred to the Western College of Veterinary Medicine for treatment. Eight bulls had unilateral and six had bilateral vesicular adenitis. The most common isolate was Arcanobacterium pyogenes. Corynebacterium pseudotuberculosis was isolated from one bull. Bulls were subjected to rectal palpation and ultrasonography of the vesicular glands, semen collection by electroejaculation, and intraglandular treatment with ceftiofur (n=13) and if necessary, a second intraglandular treatment of penicillin (n=6). One bull was treated only with an initial intraglandular injection of penicillin. Bulls were evaluated once a week over 6 weeks by palpation of the glands, and evaluation of semen. All bulls recovered from vesicular adenitis after 3-6 weeks. There was a difference in the amount of pus (P=0.042), leukocytes (P<0.001) and blood (P=0.003) present in ejaculates from before treatment to 3 weeks after treatment. Pixel intensities in ultrasonographic images of healthy or affected vesicular glands, whether treated or untreated, did not change over time. Intraglandular injection of ceftiofur in yearling bulls via the ischiorectal fossa was effective for treating vesicular adenitis.

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A study was conducted to characterize the gross and cytological characteristics of secretions from normal canine anal sacs. Fifty dogs with no recent history of anal-sac disease were selected consecutively and their anal sacs manually expressed. Of the 47 dogs with known or estimated ages, 27 were < 1 year olds. However, these youngest dogs were not significantly different from the rest in cell counts, viscosity, or presence of dark or light solid portions of the secretions. For all dogs combined, the secretions typically contained 41 (median) corneocytes per 400x magnification microscopic field, lots of basophilic background debris, and a mixture of Gram-positive cocci (173 per field; median), Gram-positive rods (29 per field; median), and Gram-negative rods (two per field; median). Although median counts across 10 400x fields per sac indicated no leucocytes or yeasts, many of the normal dogs had an occasional 400x field with non-degenerate neutrophils and yeasts. Erythrocytes were not a feature of these anal-sac secretions, and mononuclear leucocytes were found only rarely. Extreme heterogeneity existed in gross colour, consistency and the amount of solid material. Thus, these latter parameters probably would not be valid indicators of anal-sac disease.

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OBJECTIVE: To quantify numbers of leucocytes, keratinocytes and microorganisms in, as well as the turgidity, colour and consistency of, anal sac exudates in clinically normal dogs. DESIGN: Selection criteria were formed based on the absence of clinical signs associated with anal sac disease, and the absence of factors potentially affecting colonic flora. Anal sacs were palpated for turgidity then expressed onto a swab, where colour and consistency were noted. A squash preparation made from any exudate was heat fixed and stained with modified Wright's stain. Eight representative 1000x oil immersion fields from each were examined for leucocytes, erythrocytes, keratinocytes, bacteria and yeast. Results were summarised, and cytological counts grouped into quartiles (minimal, few, moderate, numerous). Due to the multivariate nature of the study and limited subject numbers, further significant statistical analysis could not be performed. RESULTS: Seventeen dogs satisfied the selection criteria. The physical characteristics of the exudate and sac varied, though 31/34 sacs were empty or soft, 22/27 exudates were light or dark brown and 19/27 exudates were a thin liquid. Total leucocyte, keratinocyte and bacilli counts were extremely variable. Yeasts were present in 26/208 microscopic fields examined cytologically. Only 5/208 fields showed numerous cocci. A single instance of intracellular bacteria and a single erythrocyte were noted following examination of all fields. CONCLUSION: In this study, the characteristics of normal anal sacs and their exudate varied but greater than 70% showed similar features. Exudate cytology was highly variable, though yeasts were uncommon, and intracellular cocci and erythrocytes extremely rare.

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Macroscopic and cytological aspects of anal sac content were evaluated in 40 normal dogs and 10 dogs each with pyoderma, Malassezia dermatitis associated with atopic dermatitis and uncomplicated atopic dermatitis. Bacteria isolated from anal sacs were compared with those from abdominal skin and hair in 20 normal dogs and 10 dogs with pyoderma. There was no difference between the groups in anal sac dimension, or in the colour, consistency or presence of granules in their content. Extracellular bacteria were found in higher numbers in diseased animals, whereas intracellular bacteria were observed in 40% of dogs with pyoderma and in only 2.5% of normal dogs. Malassezia spp. were present in 15.7% of dogs, with no difference between groups. Neutrophils were observed in 12.5% of normal dogs, 30% of dogs with Malassezia dermatitis with underlying atopic dermatitis and in 70 and 80% of dogs with pyoderma and uncomplicated atopic dermatitis, respectively. Seven bacterial species were isolated from anal sacs, with no difference between normal dogs and dogs with pyoderma. In five normal animals and in four dogs with pyoderma the same bacterial strains were isolated from anal sacs and from abdominal skin and hair.

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