TWO NEW CERATOMYXA SPECIES (MYXOSPOREA: CERATOMYXIDAE) INFECTING THE GALL BLADDER OF MARINE FISHES FROM THE SOUTH-CENTRAL COAST OF VIETNAM.

Myxospores discovered floating free in the bile of marine fishes from the south-central coast of Vietnam were identified using morphological and molecular methods, leading to the description of 2 new species. Ceratomyxa chauvanminhi n. sp. was detected in 16% (8/50) of cultured barramundi Lates calcarifer (Bloch) specimens, and Ceratomyxa sekoi n. sp. was found in 20% (5/25) of wild largehead hairtail Trichiurus lepturus Linnaeus specimens. The spores of C. chauvanminhi n. sp. are very shallowly ovoid, slightly crescent shaped, and 11.5 ± 0.5 (10.7-12.4) µm thick, 5.8 ± 0.2 (5.4-6.1) µm long, and 5.5 ± 0.2 (5.2-5.7) µm wide. Their posterior angles are slightly concave at 158.7° ± 4.2° (151.3°-164.8°), and they possess 2 equal spherical polar capsules 2.5 ± 0.2 (2.1-2.9) µm in diameter. The spores of C. sekoi n. sp. are 5.6 ± 0.2 (5.0-6.1) µm long, 75.5 ± 4.8 (68.9-90.0) µm thick, and 5.5 ± 0.1 (5.4-5.6) µm wide, with 2 equal, slightly anterior spherical polar capsules 2.1 ± 0.2 (1.7-2.4) µm in diameter. Although C. sekoi n. sp. spores resemble those of species of MyxodavisiaZhao, Zhou, Kent, and Whipps, 2008, characterized by long tapering valves, genetic analyses distinctly place this new species within the Ceratomyxa Thélohan, 1892 lineage. This study contributes to the understanding myxosporean diversity in Vietnamese waters and highlights the difficulty associated with distinguishing between the genera Ceratomyxa and Myxodavisia.

Examination of the thickness of the different components of gallbladder wall in cases with mucocele and/or cholecystitis.

OBJECTIVE: To assess the thickness of each layer of the gallbladder wall with different diseases in dogs. SAMPLE: 72 gallbladders. METHODS: Retrospective study of dogs that underwent cholecystectomy. Histopathological specimens of the gallbladders were reviewed. Histopathological diagnosis was made as gallbladder mucocele or cholecystitis, and cholecystitis was further categorized into chronic cholecystitis, acute-on-chronic cholecystitis, acute cholecystitis, and necrotic cholecystitis. The thickness of each layer of the gallbladder wall was measured. RESULTS: 22 dogs were diagnosed with gallbladder mucocele without cholecystitis, 24 with gallbladder mucocele and cholecystitis, 20 with only cholecystitis, and 6 as normal. Histopathological subclassification of cholecystitis in 44 gallbladders led to diagnosis of chronic cholecystitis in 21 gallbladders, acute-on-chronic cholecystitis in 10 gallbladders, acute cholecystitis in 6 gallbladders, and necrotic cholecystitis in 7 gallbladders. The thickness of the entire wall of the gallbladder (P < .0001) and the thickness of the mucosa (P < .0001) and subserosa (P < .0001) were affected by the different disease processes. CLINICAL RELEVANCE: Layers of the gallbladder wall were affected by diseases present in the gallbladder. It resulted in a difference in the thickness of the wall of the gallbladder among the gallbladder diseases in this study. Histopathological changes should be taken into consideration before surgery while deciding what technique to use to perform a cholecystectomy.

Characteristics of gallbladder microbiome in healthy dogs and cats, dogs with gallbladder mucocele, and cats with suspected cholangitis/cholangiohepatitis.

The aim of this study was to investigate and characterize the microbiome in bile samples obtained from dogs with gallbladder mucocele (6), cats with suspected cholangitis/cholangiohepatitis (4), as well as from healthy dogs (6) and cats (4). Our goal was to compare the microbiome patterns with clinical findings and bacterial culture results in diseases of the gallbladder and to identify a potential microbial biomarker of diseased groups. The microbial taxa composition revealed that Proteobacteria were the most dominant phylum in healthy and diseased individuals in all groups. Individuals from six families including Burkholderiaceae, Phyllobacteriaceae, Bradyrhizobiaceae, Sphingomonadaceae, Moraxellaceae, and Caulobacteraceae, constituted the core microbiome in the gallbladder of healthy dogs. A combination of LEfSe analysis and Taxa2ASV decomposer revealed that Pseudomonaceae and Ruminococcaceae exclusively occurred in the mucocele group. In conclusion, this study determined the core microbiome in the gallbladder of healthy dogs and the possible biomarkers (Pseudomonaceae and Ruminococcaceae) of gallbladder mucocele in dogs.

Le but de cette étude était d'étudier et de caractériser le microbiome dans des échantillons de bile obtenus auprès de chiens atteints de mucocèle de la vésicule biliaire (6), de chats suspectés de cholangite/cholangiohépatite (4), ainsi que de chiens (6) et de chats en bonne santé (4). Notre objectif était de comparer les modèles de microbiome avec les résultats cliniques et les résultats de cultures bactériennes dans les maladies de la vésicule biliaire et d'identifier un biomarqueur microbien potentiel des groupes malades. La composition des taxons microbiens a révélé que les protéobactéries constituaient le phylum le plus dominant chez les individus sains et malades de tous les groupes. Des individus de six familles, dont Burkholderiaceae, Phyllobacteriaceae, Bradyrhizobiaceae, Sphingomonadaceae, Moraxellaceae et Caulobacteraceae, constituaient le microbiome central de la vésicule biliaire de chiens en bonne santé. Une combinaison de l'analyse LEfSe et du décomposeur Taxa2ASV a révélé que les Pseudomonaceae et les Ruminococcaceae étaient exclusivement présentes dans le groupe des mucocèles. En conclusion, cette étude a déterminé le microbiome central de la vésicule biliaire de chiens en bonne santé et les biomarqueurs possibles (Pseudomonaceae et Ruminococcaceae) de la mucocèle de la vésicule biliaire chez le chien.(Traduit par Docteur Serge Messier).

Increased lipogenesis and lipidosis of gallbladder epithelium in dogs with gallbladder mucocele formation.

BACKGROUND: Gallbladder disease in people is frequently associated with disorders of lipid metabolism and metabolic syndrome. A recently emergent gallbladder disease of dogs, referred to as mucocele formation, is characterized by secretion of abnormal mucus by the gallbladder epithelium and is similarly associated with hyperlipidemia, endocrinopathy, and metabolic dysfunction. The cause of gallbladder mucocele formation in dogs is unknown. METHODS: A prospective case-controlled study was conducted to gain insight into disease pathogenesis by characterization of plasma lipid abnormalities in 18 dogs with gallbladder mucocele formation and 18 age and breed matched control dogs using direct infusion mass spectrometry for complex plasma lipid analysis. This analysis was complemented by histochemical and ultrastructural examination of gallbladder mucosa from dogs with gallbladder mucocele formation and control dogs for evidence of altered lipid homeostasis of the gallbladder epithelium. RESULTS: Gallbladder mucocele formation in dogs carried a unique lipidomic signature of increased lipogenesis impacting 50% of lipid classes, 36% of esterified fatty acid species, and 11% of complex lipid species. Broad enrichment of complex lipids with palmitoleic acid (16:1) and decreased abundance within complex lipids of presumptive omega-3 fatty acids eicosapentaenoic (20:5) and docosahexaenoic (22:6) was significant. Severe lipidosis of gallbladder epithelium pinpoints the gallbladder as involved causally or consequently in abnormal lipid metabolism. CONCLUSION: Our study supports a primary increase in lipogenesis in dogs with mucocele formation and abnormal gallbladder lipid metabolism in disease pathogenesis.

Determining the patency of biliary tracts in dogs with gallbladder mucocele using near-infrared cholangiography with indocyanine green.

Cholecystectomy is indicated for gallbladder mucoceles (GBM). Evaluating the patency of the biliary duct and precise biliary tree visualization is crucial for reducing the risk of compromised bile flow after surgery. Therefore, intraoperative cholangiography (IOC) is recommended during cholecystectomy to prevent biliary tract injury. Although indocyanine green (ICG) cholangiography has been extensively reported in human medicine, only one study has been conducted in veterinary medicine. Therefore, this study aimed to demonstrate the use of ICG for IOC to identify fluorescent biliary tract images and determine the patency of the common bile duct during cholecystectomy in dogs. This study comprised 27 dogs, consisting of 17 with gallbladder mucoceles (GBM) and 10 controls, specifically including dogs that had undergone elective cholecystectomy for GBM. ICG injection (0.25 mg/kg) was administered intravenously at least 45 minutes before surgery. During the operation, fluorescent images from cholangiography were displayed on the monitor and obtained in black-and-white mode for the comparison of fluorescence intensity (FI). The FI values of the gallbladders (GBs) and common bile duct (CBD) were measured using FI analyzing software (MGViewer V1.1.1, MetapleBio Inc.). The results demonstrated successful CBD patency identification in all cases. Mobile GBM showed partial gallbladder visibility, whereas immobile GBM showed limited visibility. Additionally, insights into the adequate visualization of the remaining extrahepatic biliary tree anatomy were provided, extending beyond the assessment of CBD patency and gallbladder intensity. Our study demonstrates the potential of fluorescent IOC using intravenous injection of ICG for assessing the patency of the cystic duct and common bile duct during cholecystectomy in patients with GBM, eliminating the need for surgical catheterization and flushing of the biliary ducts. Further research is warranted to investigate and validate the broader applicability of ICG cholangiography in veterinary medicine.

Concurrent hepatopathy in dogs with gallbladder mucocele: Prevalence, predictors, and impact on long-term outcome.

BACKGROUND: Information is limited regarding the prevalence and importance of hepatic histologic abnormalities in dogs with gallbladder mucocele (GBM). OBJECTIVES: To (a) report prevalence of hepatic histologic abnormalities in dogs with GBM (b) evaluate for association between hepatic abnormalities and outcome in dogs with GBM (c) evaluate whether neutrophil-to-lymphocyte ratio (NLR) differs in dogs with GBM with and without specific hepatic lesions. ANIMALS: Fifty-two dogs with grossly and histologically confirmed GBM. METHODS: Multicenter, retrospective study of dogs with GBM undergoing cholecystectomy with concurrent liver biopsy. Archived histological sections of gallbladder and liver evaluated by investigators blinded to data. Proportions of dogs with each histologic abnormality alive vs deceased at 1, 3, and 12 months post-cholecystectomy compared. Mann-Whitney U performed to determine if NLR differed in dogs with or without selected lesions. RESULTS: 51/52 (98%, 95% CI [89%, 99%]) dogs with GBM had at least 1 hepatic histologic abnormality. Hepatic fibrosis (37/51; 73%, 95% CI [59%, 83%]), biliary hyperplasia (29/52; 56%, 95% CI [42%, 68%]), and portal inflammation (25/52; 48%, 95% CI [35%, 61%]) were most common. The proportion of dogs alive vs dead differed based on the fibrosis score at 1, 3, and 12 (P ≤ .04) months post-cholecystectomy. Dogs with hepatic necrosis (P = .006) and cholangitis/cholangiohepatitis (P = .02) had higher NLRs compared to dogs without these lesions. CONCLUSIONS AND CLINICAL IMPORTANCE: Histologic abnormalities of the liver are common in dogs with GBM. A higher portal fibrosis score might be associated with shortened long-term survival after cholecystectomy for dogs with GBM. An increase in NLR might predict hepatic necrosis and cholangitis/cholangiohepatitis in dogs with GBM.

MUC5AC and MUC5B expression in canine gallbladder mucocele epithelial cells.

Gallbladder mucocele (GBM) is one of the most common gallbladder diseases in dogs. Its pathogenesis has not yet been clarified, but excessive accumulation of a secretory gel-forming mucin, MUC5AC in the gallbladder has been reported. This study aimed to ascertain if MUC5AC overproduction resulted in mucus accumulation in the gallbladder during GBM development. Eleven dogs undergoing cholecystectomy who were pathologically diagnosed with GBM were included, and the expression level of mucins, particularly MUC5AC and MUC5B, in their gallbladder epithelial cells was compared with those in normal gallbladder epithelial cells. On reverse transcription-quantitative polymerase chain reaction screening, there was a significant difference (P<0.05) in the mRNA expression level of MUC1, but not of other mucins including MUC5AC and MUC5B, between mucocele and normal gallbladder epithelial cells. Protein expression levels were also evaluated for MUC5AC and MUC5B using immunohistochemistry. There was little immunoreactivity for MUC5AC, whereas MUC5B showed definitive staining in gallbladder epithelial cells. There was no difference in MUC5AC and MUC5B protein expression levels between mucocele and normal gallbladder epithelial cells. These data suggest that excessive production of mucin, especially MUC5AC and MUC5B, does not occur in canine GBM, and that abnormal mucus excretion, rather than excessive mucus production, may be the cause of GBM development.

Increasing age and severe intraoperative hypotension associated with nonsurvival in dogs with gallbladder mucocele undergoing cholecystectomy.

OBJECTIVE: To identify prognostic indicators and inflammatory markers associated with nonsurvival in dogs with gallbladder mucoceles (GBMs) following cholecystectomy and to evaluate C-reactive protein (CRP) and haptoglobin concentrations in dogs with GBMs compared to healthy controls. ANIMALS: 25 dogs that underwent cholecystectomy for removal of GBM and 20 healthy control dogs. METHODS: A prospective, multicenter cohort study. Survival outcomes to hospital discharge and 2 weeks postdischarge were recorded from medical records. Laboratory variables, inflammatory markers (CRP and haptoglobin), and 25-hydroxyvitamin(OH) D (25[OH]D) concentrations were measured preoperatively. Associations between signalment, clinicopathologic variables, acute patient physiologic and laboratory evaluation (APPLEFAST) scores, inflammatory markers, 25(OH)D concentration, and survival were analyzed using logistic regression. RESULTS: 76% (19/25) and 68% (17/25) of dogs survived to hospital discharge and 2 weeks postdischarge, respectively. For each additional year of age, the odds of nonsurvival in hospital and 2 weeks postdischarge increased by 2.2 (P = .01; 95% CI, 1.2 to 5.0) and 1.7 (P = .04; 95% CI, 1.0 to 3.2), respectively. Intraoperative systolic blood pressure ≤ 65 mm Hg increased the probability of nonsurvival in hospital (P < .04). Gallbladder perforation, APPLEFAST scores, and preoperative serum concentrations of CRP, haptoglobin, and 25(OH)D were not associated with survival. Serum CRP and haptoglobin concentrations were greater in dogs with GBM compared to controls (P < .001). CLINICAL RELEVANCE: Increasing age and intraoperative systolic blood pressure ≤ 65 mm Hg were associated with nonsurvival in dogs with GBM undergoing cholecystectomy. Serum CRP, haptoglobin, and 25(OH)D were not associated with nonsurvival postcholecystectomy in this sample population.

Ultrasonographic measurement of gallbladder wall thickness in fasted dogs without signs of hepatobiliary disease.

BACKGROUND: Ultrasound-determined gallbladder wall thickness is widely used to aid in the diagnosis of gallbladder disease, but no reference values supported by published measurement data are available in dogs. HYPOTHESIS/OBJECTIVE: Establish normal thickness of the gallbladder wall in dogs. ANIMALS: Fifty-three dogs presented to a referral hospital and required abdominal ultrasound examination for reasons unrelated to primary hepatobiliary disease. METHODS: Cross-sectional observational study recruiting dogs requiring abdominal ultrasound examination. A standard sequence of gallbladder wall images was recorded for later review. Inclusion criteria were normal ultrasonographic hepatobiliary, pancreatic, and small intestinal findings. Exclusion was determined by 2 European College of Veterinary Internal Medicine (ECVIM)-certified veterinary internists blinded to gallbladder wall thickness data. Dogs were excluded if they had inadequate medical records, a previous history of hepatobiliary, gastrointestinal, or pancreatic disease likely to impact the biliary system (eg, chronic vomiting, nausea, jaundice, diarrhea), unexplained increases in liver enzyme activities, hypoalbuminemia, or ascites. Gallbladder wall thickness was determined by 2 European College of Veterinary Diagnostic Imaging (ECVDI)-certified veterinary radiologists working together to generate a consensus for each dog. The final output was the maximum normal wall thickness for this population of dogs. RESULTS: The upper limit for gallbladder wall thickness in 53 fasted (8 hours) dogs <40 kg was 1.30 mm (90% confidence interval, 1.19-1.41). CONCLUSIONS AND CLINICAL IMPORTANCE: Normal gallbladder wall thickness in dogs is lower than previously reported. Additional studies are required to determine potential effects of body weight and the optimal cut-off to distinguish between healthy and diseased gallbladders.

Ultrasonographic features of gallbladder wall thickening in dogs with hypoalbuminemia.

Hypoalbuminemia has been considered as a cause of gallbladder wall thickening in humans and dogs. Recent study revealed that gallbladder wall thickening in dogs with hypoalbuminemia may not be associated with serum albumin/plasma levels within 48 h of ultrasound study. However, gallbladder wall edema may change within 48 h, and the ultrasonographic features of gallbladder wall thickening in dogs with hypoalbuminemia have not been reported. The purpose of this study is to describe the relationship between serum albumin levels within 24 h of ultrasound and gallbladder wall thickening, and to describe the ultrasonographic features of thickened gallbladder walls in dogs with hypoalbuminemia. 37 hypoalbuminemic dogs with gallbladder ultrasound images were retrospectively included. Ultrasound studies were reviewed, and gallbladder wall thickness, layering appearance, echogenicity, echotexture, distribution, evidence of gallbladder mucocele, and presence of peritoneal effusion were recorded. Additionally, serum albumin levels within 24 h of ultrasound study and the administerd sedation were recorded. The prevalence of gallbladder wall thickening in dogs with hypoalbuminemia was 13.5%. The 3-layer appearance of the gallbladder wall was observed in 4 dogs, and a single-layer gallbladder wall thickening in one dog. Diffuse thickening was observed in all 5 dogs. The serum albumin level of dogs with gallbladder wall thickening was not different (p = 0.14) from dogs without thickening. Gallbladder wall thickening was not common, occurring only with mild hypoalbuminemia, and was commonly associated with a 3-layer appearance and considered as gallbladder wall subserosal edema. Causes other than hypoalbuminemia may be responsible for thickening of the gallbladder wall in dogs with hypoalbuminemia.

Evaluation of gallbladder motility assessed by ultrasonography in dogs with hyperlipidemia.

BACKGROUND: The pathogenesis of gallbladder (GB) mucoceles in dogs is unknown. It has been proposed that hyperlipidemia could impair GB motility and contribute to GB mucocele formation. HYPOTHESIS/OBJECTIVES: The objective of this study was to compare GB motility in dogs with hyperlipidemia to control dogs using ultrasonography. We hypothesized that hyperlipidemic dogs will have decreased GB motility compared with controls. ANIMALS: Twenty-six hyperlipidemic and 28 healthy, age-matched control dogs were prospectively enrolled. METHODS: Cholesterol and triglyceride concentrations were measured in all dogs. Hyperlipidemia was defined as hypercholesterolemia (>332 mg/dL) and/or hypertriglyceridemia (>143 mg/dL) using a biochemical analyzer. Ultrasound was performed before feeding, and 60 and 120 minutes after ingestion of a high fat diet. Gallbladder volumes (GBV) and ejection fractions (EF) were calculated. RESULTS: Hyperlipidemic dogs had significantly larger GBVs (ml/kg) before feeding and 60 minutes after feeding of 1.2 (0.4-7.5; P = .008) and 0.6 (0.1-7.2; P = .04) compared with controls 0.6 (0.2-2.6) and 0.4 (0.1-1.9), respectively. Severely hyperlipidemic dogs had significantly larger GBV at baseline, 60 minutes, and 120 minutes of 1.7 (0.6-7.5; P = .03), 1.3 (0.4-7.2; P = .02), and 1.3 (0.2-8.2; P = .04), respectively compared with mildly hyperlipidemic dogs. EFs at 60 and 120 minutes between controls, hyperlipidemic, and severely hyperlipidemic were all 0.3 at 60 minutes and 0.5, 0.3, and 0.3 at 120 minutes, respectively which were not statistically different. CONCLUSIONS AND CLINICAL IMPORTANCE: Hyperlipidemia leads to GB distention in dogs which could lead to retention of bile and gallbladder disease.

The effect of flushing of the common bile duct on hepatobiliary markers and short-term outcomes in dogs undergoing cholecystectomy for the management of gall bladder mucocele: A randomized controlled prospective study.

OBJECTIVE: To determine the effect of flushing of the common bile duct (CBD) on hepatobiliary markers and short-term outcome in dogs undergoing cholecystectomy for the management of gallbladder mucocele (GBM). STUDY DESIGN: Randomized, controlled, prospective study. ANIMALS: Thirty-two client-owned dogs. METHODS: Dogs were allocated randomly to either a "flush" group or a "non-flush group." Flushing was performed in a normograde fashion, followed by a routine cholecystectomy. Data collected included presenting clinical signs, preoperative and 3-day postoperative hepatobiliary markers (alkaline phosphatase, ALP; alanine aminotransferase, ALT; gamma glumatyl-transferase, GGT; bilirubin; cholesterol; triglycerides), duration of hospitalization, and complications. These data were compared between groups. RESULTS: Sixteen dogs were enrolled in each group. One dog (in the flush group) was excluded following diagnosis of hepatic lymphoma. Border terriers were overrepresented (20/31). Overall, there were marked reductions from preoperative to 3 days postoperative in serum bilirubin (p = .004), ALP (p = .020), ALT (p < .001), GGT (p = .025), and cholesterol (p < .001) values. There was no difference in any marker between groups. Survival to discharge was 90.3% (28/31 dogs). CONCLUSION: Cholestatic markers decreased significantly 3 days postcholecystectomy. No short-term clinical or clinico-pathological benefits were identified when flushing the CBD in dogs undergoing cholecystectomy for GBM. CLINICAL SIGNIFICANCE: The findings of the study do not support routine flushing of the CBD during cholecystectomy for GBM in dogs.

Ultrasonographic Appearance of Gallbladder Neoplasia in 14 Dogs and 1 Cat.

Ultrasonography is the most commonly used diagnostic imaging tool for gallbladder disease in veterinary medicine. Primary gallbladder neoplasia is an uncommon finding with variable prognosis for which no studies have been published describing their ultrasonographic appearance and diagnosis. This retrospective, multicenter, case series study examines the ultrasonographic appearance of gallbladder neoplasia with histologically or cytologically confirmed diagnoses. A total of 14 dogs and 1 cat were analyzed. All discrete masses were sessile in shape and varied in size, echogenicity, location, and gallbladder wall thickening. All studies with images showing Doppler interrogation exhibited vascularity. Cholecystoliths were an uncommon finding, being present in only one case in this study, unlike in humans. The final diagnosis of the gallbladder neoplasia was neuroendocrine carcinoma (8), leiomyoma (3), lymphoma (1), gastrointestinal stromal tumor (1), extrahepatic cholangiocellular carcinoma (1), and adenoma (1). Findings from this study indicate that primary gallbladder neoplasms have variable sonographic appearances and cytologic and histologic diagnoses.

Increased radiographic opacity in the region of the gallbladder can be identified in dogs with large amounts of gallbladder sediment, sludge balls, and gallbladder mucocele.

Increased soft-tissue opacity in the region of the canine gallbladder is incidentally detected on radiographs. We hypothesized that there is a difference in the detection of gallbladder sediment on radiographs depending on the amount or mobility of the sediment. In this retrospective and analytical study, we aimed to assess the ultrasonographic features of gallbladder sediment that were detected radiographically. We also aimed to assess the differences in the detection of increased opacity of the gallbladder between radiographic views. We included 223 dogs that underwent thoracic radiography, abdominal radiography, and gallbladder ultrasonography. Ultrasonographic images of the gallbladder were divided into five groups: group 1, gravity-dependent sediment occupying < 50% of the gallbladder; group 2, gravity-dependent sediment occupying ≥50%; group 3, sediment attached to the gallbladder wall; group 4, sludge ball; and group 5, gallbladder mucocele. Dogs showing increased opacity on subjective assessment of any radiographic view were recorded, and the sensitivity of radiographic views for detecting gallbladder sediment was analyzed. Of 168 dogs with gallbladder sediment, 37 had increased opacity on at least one radiographic projection. The frequency was compared as a percentage within each category, and Group 4 was the highest percentage with increased radiographic gallbladder opacity, followed by Groups 2 and 5. The sensitivity for detecting increased opacity was highest in the thoracic ventrodorsal view. Thus, in dogs with increased gallbladder opacity on radiographs, large amounts of gallbladder sediment, sludge balls, and gallbladder mucocele should be considered differential diagnoses. In addition, the thoracic ventrodorsal view is recommended to evaluate gallbladder opacity.

Gallbladder microbiota in healthy dogs and dogs with mucocele formation.

To date studies have not investigated the culture-independent microbiome of bile from dogs, a species where aseptic collection of bile under ultrasound guidance is somewhat routine. Despite frequent collection of bile for culture-based diagnosis of bacterial cholecystitis, it is unknown whether bile from healthy dogs harbors uncultivable bacteria or a core microbiota. The answer to this question is critical to understanding the pathogenesis of biliary infection and as a baseline to exploration of other biliary diseases in dogs where uncultivable bacteria could play a pathogenic role. A pressing example of such a disease would be gallbladder mucocele formation in dogs. This prevalent and deadly condition is characterized by excessive secretion of abnormal mucus by the gallbladder epithelium that can eventually lead to rupture of the gallbladder or obstruction of bile flow. The cause of mucocele formation is unknown as is whether uncultivable, and therefore unrecognized, bacteria play any systematic role in pathogenesis. In this study we applied next-generation 16S rRNA gene sequencing to identify the culture-negative bacterial community of gallbladder bile from healthy dogs and gallbladder mucus from dogs with mucocele formation. Integral to our study was the use of 2 separate DNA isolations on each sample using different extraction methods and sequencing of negative control samples enabling recognition and curation of contaminating sequences. Microbiota findings were validated by simultaneous culture-based identification, cytological examination of bile, and fluorescence in-situ hybridization (FISH) performed on gallbladder mucosa. Using culture-dependent, cytological, FISH, and 16S rRNA sequencing approaches, results of our study do not support existence of a core microbiome in the bile of healthy dogs or gallbladder mucus from dogs with mucocele formation. Our findings further document how contaminating sequences can significantly contribute to the results of sequencing analysis when performed on samples with low bacterial biomass.

Laparoscopic cholecystectomy using the subserosal layer dissection technique in dogs: 34 cases (2015-2021).

OBJECTIVES: To describe a standardised subserosal layer dissection technique and evaluate its outcomes in canine laparoscopic cholecystectomy. MATERIALS AND METHODS: Medical records of dogs undergoing laparoscopic cholecystectomy using the standardised subserosal layer dissection technique for the treatment of cholecystolithiasis, cholecystitis, and gall bladder mucocele at a single veterinary hospital from January 2015 to September 2021 were extracted. Operative time, subserosal layer dissection achievement rate, open conversion rate, and complication rate were evaluated. RESULTS: Thirty-four dogs were included. The most common preoperative diagnosis was cholecystolithiasis (n=29). Operative time was 190 minutes (range: 110 to 330 minutes). Subserosal layer dissection of more than 90% of the gall bladder bed was achieved in 27 (79%) dogs. Conversion to open surgery was required in three (8.8%) dogs. There were no cases of intraoperative bleeding, bile duct injury, or reoperation. CLINICAL SIGNIFICANCE: This study showed that laparoscopic cholecystectomy using the standardised subserosal layer dissection technique could be performed successfully in dogs. Future prospective clinical studies are needed to determine safety and effectiveness of this technique compared to standard techniques.

Comparative evaluation of three laparoscopic cholecystectomy techniques in rabbit's model

Purpose: The aim of this randomized study was to compare the complications and perioperative outcome of three different techniques of laparoscopic cholecystectomy (LC). Changes in the liver function test after LC techniques were investigated. Also, we compared the degree of postoperative adhesions and histopathological changes of the liver bed. Methods: Thirty rabbits were divided into three groups: group A) Fundus-first technique by Hook dissecting instrument and Roeder Slipknot applied for cystic duct (CD) ligation; group B) conventional technique by Maryland dissecting forceps and electrothermal bipolar vessel sealing (EBVS) for CD seal; group C) conventional technique by EBVS for gallbladder (GB) dissection and CD seal. Results: Group A presented a longer GB dissection time than groups B and C. GB perforation and bleeding from tissues adjacent to GB were similar among tested groups. Gamma-glutamyl transferase and alkaline phosphatase levels increased (p ≤ 0.05) on day 3 postoperatively in group A. By the 15th postoperative day, the enzymes returned to the preoperative values. Transient elevation of hepatic transaminases occurred after LC in all groups. Group A had a higher adherence score than groups B and C and was associated with the least predictable technique. Conclusions: LC can be performed using different techniques, although the use of EBVS is highly recommended.

High frequency of cholecystitis in dogs with gallbladder mucocoele in Hong Kong.

The aims of this retrospective study were to characterise the epidemiological, clinical, histopathological, and microbiological findings as well as surgical outcomes in dogs admitted to a specialist veterinary hospital in Hong Kong for surgical management of gallbladder mucocoele (GBM). Inclusion criteria were cases with histopathological diagnosis of GBM and accompanying abdominal imaging, serum biochemistry, bile culture, and liver biopsy histology results. Fifty-six cases met the inclusion criteria. The median age at diagnosis was 12 years (range, 5-16 years). Miniature or toy pure-breed dogs were most commonly affected, including Poodles, Pomeranians, Schnauzers, Bichon frises and Chihuahuas. However, no breed was over-represented compared with their expected proportions among annual hospital admissions. Histological evidence of cholecystitis was present in 84% of cases, including acute cholecystitis in 18%, chronic cholecystitis in 37.5%, acute on chronic cholecystitis in 28% and acute with necrosis in 6%. The most common liver lesions were cholestasis in 64%, along with portal fibrosis in 55%, oedema in 50% and bile duct hyperplasia in 50%. Bile culture was positive in 29.6% of cases. Escherichia coli and Enterobacter species were most commonly isolated. Stentrophomonas maltophili was cultured from one case. Of the 16 cases where bacteria were isolated from bile culture, 94% had evidence of chronic cholecystitis and 81% had evidence of cholangiohepatitis. Fifty dogs (89.3%) survived to discharge including 5/5 dogs with ruptured gallbladders. Of 34 dogs with follow-up data, 21/34 (61.8%) were still alive 12 months later. Gallbladder mucocoeles were frequently associated with both acute and chronic inflammation. High survival rates to discharge were achieved.

Clinical relationship between histopathological necrotic/partial necrotic findings and disease condition of gallbladder mucoceles in dogs.

Gallbladder mucocele (GM) is a common extrahepatic biliary disease recognized in dogs and is defined as the expansion and extension of the gallbladder by an accumulation of semi-solid bile or bile acid. Histopathological diagnosis of necrotizing cholecystitis and transmural coagulative necrosis of the gallbladder wall shows poor prognosis. Conversely, histopathological diagnosis with partial necrotic findings is often achieved. We hypothesized that histopathological partial necrosis of the gallbladder wall is the primary lesion of necrotic cholecystitis or transmural ischemic necrosis. Therefore, we investigated the relationship between histopathological necrosis/ partial necrosis findings and their clinical conditions. We retrospectively analyzed 55 dogs diagnosed with GM that had undergone cholecystectomy at the Yamaguchi University Animal Medical Center. The group with histopathological necrosis/partial necrosis of the gallbladder wall showed elevated levels of preoperative white blood cells, alanine transaminase, alkaline phosphatase, γ-glutamyltransferase, total bilirubin, and C-reactive protein compared to the non-necrotic group. Partial necrosis of the gallbladder wall may affect the progression of the disease and hematological abnormalities. Additionally, all death cases until 2 weeks were included in the histopathological necrosis/partial necrosis group. In this study, we found that poor prognosis factors were associated with partial necrosis of the gallbladder wall. Furthermore, these cases of partial necrosis showed elevated levels of blood test parameters. These results suggest that necrosis of the gallbladder wall is associated with poor prognosis and poor pathophysiological conditions.

Screening of bacterial DNA in bile sampled from healthy dogs and dogs suffering from liver- or gallbladder-associated disease.

Although the biliary system is generally aseptic, gallbladder microbiota has been reported in humans and some animals apart from dogs. We screened and analyzed the bacterial deoxyribonucleic acid in canine gallbladders using bile sampled from 7 healthy dogs and 52 dogs with liver- or gallbladder-associated disease. PCR screening detected bacteria in 17.3% of diseased dogs (9/52) and none in healthy dogs. Microbiota analysis of PCR-positive samples showed that the microbial diversity differed between liver- and gallbladder-associated disease groups. Thus, a specific bacterial community appears to occur at a certain frequency in the bile of diseased dogs.