Molecular-phylogenetic investigation of trichomonads in dogs and cats reveals a novel Tritrichomonas species.

BACKGROUND: Trichomonosis is a common infection in small animals, mostly manifesting in gastrointestinal symptoms such as diarrhea. Although oral trichomonads are also known, the species found colonizing the large intestine are more frequently detected protozoa. METHODS: In the present study, four wildcats, 94 domestic cats, and 25 dogs, originating from 18 different locations in Hungary, were investigated for the presence of oral and large intestinal trichomonads based on the 18S rRNA gene and ITS2. RESULTS: All oral swabs were negative by polymerase chain reaction (PCR). However, Tritrichomonas foetus was detected in a high proportion among tested domestic cats (13.8%) and dogs (16%), and Pentatrichomonas hominis only in two domestic cats. In addition, a novel Tritrichomonas genotype was identified in one cat, probably representing a new species that was shown to be phylogenetically most closely related to Tritrichomonas casperi described recently from mice. All positive dogs and half of the positive cats showed symptoms, and among cats, the most frequent breed was the Ragdoll. CONCLUSIONS: With molecular methods, this study evaluated the prevalence of oral and intestinal trichomonads in clinical samples of dogs and cats from Hungary, providing the first evidence of T. foetus in dogs of this region. In contrast to literature data, P. hominis was more prevalent in cats than in dogs. Finally, a hitherto unknown large intestinal Tritrichomonas species (closely related to T. casperi) was shown to be present in a cat, raising two possibilities. First, this novel genotype might have been a rodent-associated pseudoparasite in the relevant cat. Otherwise, the cat was actually infected, thus suggesting the role of a predator-prey link in the evolution of this trichomonad.

Determination of Bile Acids in Canine Biological Samples: Diagnostic Significance.

The comprehensive examination of bile acids is of paramount importance across various fields of health sciences, influencing physiology, microbiology, internal medicine, and pharmacology. While enzymatic reaction-based photometric methods remain fundamental for total BA measurements, there is a burgeoning demand for more sophisticated techniques such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) for comprehensive BA profiling. This evolution reflects a need for nuanced diagnostic assessments in clinical practice. In canines, a BA assessment involves considering factors, such as food composition, transit times, and breed-specific variations. Multiple matrices, including blood, feces, urine, liver tissue, and gallbladder bile, offer insights into BA profiles, yet interpretations remain complex, particularly in fecal analysis due to sampling challenges and breed-specific differences. Despite ongoing efforts, a consensus regarding optimal matrices and diagnostic thresholds remains elusive, highlighting the need for further research. Emphasizing the scarcity of systematic animal studies and underscoring the importance of ap-propriate sampling methodologies, our review advocates for targeted investigations into BA alterations in canine pathology, promising insights into pathomechanisms, early disease detection, and therapeutic avenues.

Altered Intestinal Production of Volatile Fatty Acids in Dogs Triggered by Lactulose and Psyllium Treatment.

The intestinal microbiome of dogs can be influenced by a number of factors such as non-starch polysaccharides as well as some non-digestible oligo- and disaccharides. These molecules are only decomposed by intestinal anaerobic microbial fermentation, resulting in the formation of volatile fatty acids (VFAs), which play a central role in maintaining the balance of the intestinal flora and affecting the health status of the host organism. In the present study, the effects of lactulose and psyllium husk (Plantago ovata) were investigated regarding their influence on concentrations of various VFAs produced by the canine intestinal microbiome. Thirty dogs were kept on a standard diet for 15 days, during which time half of the animals received oral lactulose once a day, while the other group was given a psyllium-supplemented diet (in 0.67 and in 0.2 g/kg body weight concentrations, respectively). On days 0, 5, 10 and 15 of the experiment, feces were sampled from the rectum, and the concentration of each VFA was determined by GC-MS (gas chromatography−mass spectrometry). Lactulose administration caused a significant increase in the total VFA concentration of the feces on days 10 and 15 of the experiment (p = 0.035 and p < 0.001, respectively); however, in the case of psyllium supplementation, the concentration of VFAs showed a significant elevation only on day 15 (p = 0.003). Concentrations of acetate and propionate increased significantly on days 5, 10 and 15 after lactulose treatment (p = 0.044, p = 0.048 and p < 0.001, respectively). Following psyllium administration, intestinal acetate, propionate and n-butyrate production were stimulated on day 15, as indicated by the fecal VFA levels (p = 0.002, p = 0.035 and p = 0.02, respectively). It can be concluded that both lactulose and psyllium are suitable for enhancing the synthesis of VFAs in the intestines of dogs. Increased acetate and propionate concentrations were observed following the administration of both supplements; however, elevated n-butyrate production was found only after psyllium treatment, suggesting that the applied prebiotics may exert slightly different effects in the hindgut of dogs. These findings can be also of great importance regarding the treatment and management of patients suffering from intestinal disorders as well as hepatic encephalopathy due to portosystemic shunt.

Investigation of the environmental presence of multidrug-resistant bacteria at small animal hospitals in Hungary.

Multidrug-resistant bacteria can cause severe nosocomial infections in both human and veterinary clinics. The aim of this study was to investigate the presence and antibiotic susceptibility of Enterococcus, Staphylococcus and Pseudomonas strains at four small animal clinics of Hungary in 2018, as these bacteria can reliably represent the level of antimicrobial resistance in the investigated environment. A total of 177 Staphylococcus colonies were found, including 22 Staphylococcus pseudintermedius and 13 Staphylococcus aureus. As regards enterococci, 9 Enterococcus faecium, 2 E. faecalis and further 286 Enterococcus strains were isolated. The number of Pseudomonas aeruginosa isolates (n = 34) was considered too low for relevant susceptibility testing. Among staphylococci, the highest resistance was found to sulphamethoxazole (82.9%), penicillin (65.7%) and erythromycin (54.3%), while in the case of enterococci, resistance to norfloxacin and rifampicin was the most common, with 25.5% of the strains being resistant to both antibiotics. Ten methicillin-resistant S. pseudintermedius (MRSP) and six vancomycin-resistant Enterococcus (VRE) strains could be identified. Only 5.7% of the Staphylococcus isolates were susceptible to all tested agents, while this ratio was 36.2% among enterococci. The results of this study have revealed a high prevalence of antibiotic-resistant bacteria in Hungarian small animal clinics, which highlights the importance of regular disinfection processes and stringent hygiene measures in veterinary clinics.

Serum hepcidin measurements in healthy dogs using liquid chromatography/tandem mass spectrometry.

BACKGROUND: Hepcidin is a key regulator of iron homeostasis. The measurement of this hormone is essential for the diagnosis of iron deficiency anemia and might be useful as a prognostic factor in many diseases. Serum hepcidin levels have been infrequently evaluated in dogs. OBJECTIVES: The aim of this study was to measure serum hepcidin in a population of healthy dogs using liquid chromatography/tandem mass spectrometry (LC-MS/MS), considered the gold standard assay for measuring hepcidin like in human medicine. METHODS: Blood samples from 86 healthy dogs of 25 different breeds were measured with the LC-MS/MS method. Synthetic canine hepcidin was used as the standard reagent. Reference values were calculated based on the results. RESULTS: The mean hepcidin concentration of the study population was 16.6 ± 7.7 ng/mL. There reference interval (RI) was defined as 5.3-36.4 ng/mL. No significant difference was found between male and female dogs, or between different age and body weight groups. Hepcidin concentrations did not correlate with red blood cell counts, hemoglobin concentrations, iron levels, iron-binding capacities, and C-reactive protein concentrations in this healthy population. A weak negative correlation was found between hepcidin and the mean corpuscular volume. CONCLUSION: LC-MS/MS proved to be a reliable and time-effective method for the detection of canine hepcidin. The RI was similar but narrower compared with that of human studies.

Preparation of purified perikaryal and synaptosomal mitochondrial fractions from relatively small hypothalamic brain samples.

In order to measure the activity of neuronal mitochondria, a representative proof of neuronal processes, physiologically relevant mitochondrial samples need to be gained as simply as possible. Existing methods are, however, either for tissue samples of large size and/or homogenous microstructures only, or are not tested for mitochondrial function measurements. In the present article we describe a gradient fractionation method to isolate viable and well-coupled mitochondria from relatively heterogeneous histological microstructures such as the hypothalamus. With this new method, we are able to isolate a sufficient amount of functional mitochondria for determination of respiratory activity, in a short period of time, using affordable equipment. •Verified by electron microscopy, our method separates highly enriched and well-preserved perikaryal and synaptosomal mitochondria. Both fractions contain minimal cell debris and no myelin. Respiratory measurements (carried out by Clark-type electrode) confirmed undisturbed mitochondrial function providing well-evaluable records. The demonstrated protocol yields highly viable mitochondrial subfractions within 3 h from small brain areas for high-precision examinations. Using this procedure, brain regions with relatively heterogeneous histological microstructure (hypothalamus) can also be efficiently sampled.•Up to our present knowledge, our method is the shortest available procedure with the lowest sample size to gain debris-free, fully-viable mitochondria.

Expression of claudins in the normal canine gastric mucosa.

The aim of the present study was to investigate the expression pattern of claudin-1, -2, -3, -4, -5, -7, -8, -10 and -18 in the intact fundic and pyloric gastric mucosa of dogs. Intense, linear, membranous claudin-18 positivity was detected in the surface gastric cells and in the epithelial cells of the gastric glands both in the fundic and pyloric stomach regions. The mucous neck cells in the apical part of the glands, furthermore the parietal cells and chief cells of the basal part of the gland were all positive for claudin-18, in the same way as the enteroendocrine cells. Cells of the basal part of the pyloric glands showed intense, linear, membranous claudin-2 positivity, but cells of the superficial portion of these glands and the surface gastric cells in this region were claudin-2 negative. Fibroblasts, endothelial cells, lymphocytes of the propria layer, smooth muscle cells and vegetative neurons were all negative for claudin-2. All gastric epithelial cells were negative for claudin-1, -3, -4, -5, -6, -7, -8 and -10. The endothelial cells of the propria layer had intense claudin-5 positivity. We assume that claudin-18 forms a paracellular barrier against gastric acid in the healthy canine stomach, in the same way as in mice.

Cholagogue-induced gallbladder emptying in the diagnosis of canine biliary obstruction.

The efficacy of ultrasound-guided cholagogue-induced gallbladder emptying for differentiating obstructive from non-obstructive hepatobiliary diseases was studied in icteric dogs. In 7 healthy Beagle dogs, Lipofundin 20% infusion (2 ml/kg orally) evoked a vigorous gallbladder contraction of 44.2% (range: 35.3-57.6%) and proved to be a useful, well-tolerable meal for routine use. In 24 icteric dogs, gallbladder contraction was evoked by different cholagogues: the maximum reduction in gallbladder volume (%) for the three non-obstructive icteric dogs was 43.9% (range, 39.0-46.5%). The average gallbladder contraction of the 21 dogs with biliary obstruction was less than 5%. In conclusion, the stimulation of gallbladder contraction with orally applied magnesium sulphate (MgSO4) or Lipofundin can be well demonstrated by ultrasound in dogs. The examination of cholagogue-induced gallbladder emptying is a valuable technique in icteric patients to indicate surgical intervention.

Effect of enteral feeding on gallbladder function in dogs.

Nutritional support in critically ill patients is a fundamental principle of patient care. Little is known about gallbladder motility during the interdigestive phase and in response to enteral feeding. The objective of this study was to investigate the effect of enteral feeding on gallbladder function in dogs. The cholagogue meal (Lipofundin infusion) was applied in four anatomical positions (jejunum, duodenojejunal junction, descending duodenum, stomach) in five healthy Beagle dogs. Gallbladder volume (GBV) was monitored by ultrasonography. Lipofundin infusion given through the feeding tube caused a maximal gallbladder contraction of 9.2% (range 3.7-13.9%) in the jejunum, 16.5% (9.1-22.1%) at the duodenojejunal junction and 26.3% (22.8-29.5%) in the descending duodenum. When the cholagogue meal was given through the mouth, it caused a mean 33.8% (28.6-46.5%) maximum gallbladder contraction in the same animals. In conclusion, we can establish that the ultrasound-guided gallbladder emptying method proved to be a useful technique for monitoring the cholagogue effect of Lipofundin meal applied in different anatomical positions of the intestine. The deeper the position of application, the smaller and shorter gallbladder contraction was evoked.

Occurrence, clinical features and outcome of canine pancreatitis (80 cases).

Medical records of 80 dogs diagnosed with acute pancreatitis during a 4-year period were evaluated regarding history, breed predilection, clinical signs and additional examination findings. Cases were selected if compatible clinical symptoms, increased serum activity of amylase or lipase and morphologic evidence of pancreatitis by ultrasonography, laparotomy or necropsy were all present. Like in other studies, neutered dogs had an increased risk of developing acute pancreatitis. Although breed predilection was consistent with earlier reports, some notable differences were also observed. Apart from Dachshunds, Poodles, Cocker Spaniels and Fox Terriers, the sled dogs (Laikas, Alaskan Malamutes) also demonstrated a higher risk for pancreatitis according to our results. Concurrent diseases occurred in 56 dogs (70%), diabetes mellitus (n = 29, 36%) being the most common. Clinical signs of acute pancreatitis were similar to those observed in other studies. The study group represented a dog population with severe acute pancreatitis, having a relatively high mortality rate (40%) compared to data of the literature. Breed, age, gender, neutering and body condition had no significant association with the outcome. Hypothermia (p = 0.0413) and metabolic acidosis (p = 0.0063) correlated significantly with poor prognosis and may serve as valuable markers for severity assessment in canine acute pancreatitis.

A case of synchronous hepatocellular carcinoma and aortic body chemodectoma in a dog - pathological case report.

The authors describe a case of synchronously occurring (double) tumours, i.e. primary hepatocellular carcinoma and aortic body chemodectoma in a 14-year-old mixed-breed male dog. The tumours were identified during necropsy, following euthanasia. In the last months of its life, the dog showed signs of weakness, anorexia, apathy, inactivity, and abdominal palpation elicited a painful reaction. The primary liver cancer emerged in the left lateral lobe without evidence of any distant metastases. Histopathological and immunohistochemical investigations revealed a well-differentiated, trabecular, claudin-7-, claudin-5- and pancytokeratin-negative hepatocellular carcinoma. The Ki-67 proliferation index was 33%. During necropsy, a synchronously occurring benign, grade I type aortic body chemodectoma was also detected in the dog. This neuroendocrine tumour showed chromogranin-, synaptophysin-, neuron-specific enolase- and S100 protein-positivity, and the Ki-67 proliferation index was 2%. The authors believe that this is the first description of synchronously occurring hepatocellular carcinoma and aortic body chemodectoma in a dog.

Endoscopically guided nasojejunal tube placement in dogs for short-term postduodenal feeding.

OBJECTIVE: To evaluate a method for endoscopically guided nasojejunal tube placement allowing short-term postduodenal feeding and chyme withdrawal in dogs. DESIGN: Pilot study. SETTING: University teaching hospital. ANIMALS: Three healthy Beagle dogs with jejunal nipple valve fistulas. INTERVENTIONS: After the dogs were anesthetized, an 8 Fr, 250-cm polyvinyl chloride catheter was advanced through a gastroscope into the jejunum. Correct jejunal placement was established using endoscopic visualization and confirmed by fluoroscopy and radiography. The proximal end of the tube was pulled out through 1 nostril and sutured to the skin of the forehead. Thereafter, jejunal feeding was administered for 4 days. Follow-up examinations included daily confirmation of the tube's position using radiography, physical examination, and blood analyses. Withdrawal of jejunal chyme was performed after jejunal and oral feeding. MEASUREMENTS AND MAIN RESULTS: Fluoroscopic examination confirmed that endoscopic visualization alone allowed correct jejunal placement. During a 4-day postduodenal feeding period, repeated radiographic examination revealed stable positioning of the tubes within the jejunum with minor cranial displacement. The tubes were functional throughout the study without causing identifiable problems. Repeated physical examinations and blood analysis showed no abnormalities. We were able to administer the daily caloric requirements as a liquid diet. Jejunal chyme was successfully withdrawn via the tube. CONCLUSIONS: Endoscopically guided nasojejunal tube placement was shown to be a minimally invasive, well-tolerated method for short-term jejunal feeding in healthy dogs. This technique is a viable option for dogs requiring jejunal feeding but not laparotomy. The feasibility of chyme sampling is another unique application of the procedure.

Integration of woodchuck hepatitis and N-myc rearrangement determine size and histologic grade of hepatic tumors.

Integrations of woodchuck hepatitis virus (WHV) DNA and rearrangements of the N-myc 2 gene have been detected frequently in hepatocellular carcinoma (HCC) of Eastern woodchucks (Marmota monax) chronically infected with WHV. Fifty-five hepatocellular neoplasms and matched nontumor hepatic tissue specimens obtained postmortem from 13 chronic WHV carriers were analyzed and the frequency of WHV DNA integrations and of N-myc rearrangements compared in tumors of different size and histologic grade. Four small tumor nodules were classified histologically as adenomas and integrated sequences of WHV DNA were detected in two of the four tumor nodules. In one of the two nodules, there was evidence of N-myc rearrangement. Fifty-one neoplasms were classified as HCC. Seven were grade 1 HCCs. WHV DNA integrations were demonstrated in 43% but none had N-myc rearrangements. Twenty grade 2 HCCs had WHV DNA integrations in 80% and in 38% N-myc rearrangements were present. Twenty-four grade 3 HCCs had integrations of WHV DNA in 79% and N-myc rearrangements in 74%. In two other grade 3 HCCs, rearrangements of N-myc were detected in the absence of WHV DNA integrations. The 12 largest tumors in the series all were grade 2 or 3 HCCs, and in 83%, both WHV DNA integrations and N-myc rearrangements were demonstrated. In conclusion, molecular changes observed in this study suggest a progression of genetic alterations providing either a significant proliferative stimulation and/or a growth advantage in hepatocarcinogenesis of woodchucks with chronic WHV infection.

Percutaneous ultrasound-guided cholecystocentesis in dogs.

Percutaneous ultrasound-guided cholecystocentesis was performed on 13 healthy beagle dogs to determine whether percutaneous ultrasound-guided cholecystocentesis in the dog was a feasible and safe procedure. Clinical, laboratory and ultrasonographic examinations were done at 0 and 10 minutes, in the 2nd and 16th hour, and on the 7th day. They included a detailed physical examination of the mucous membranes, cardiorespiratory system and abdominal organs. Laboratory examinations of the blood consisted of a complete blood count, determination of packed cell volume (PCV), haemoglobin (Hb), total plasma protein (TPP), parameters of haemostasis including prothrombin time (PT), activated partial thromboplastin time (APTT), and enzyme activities reflecting hepatobiliary function, i.e. aspartate aminotransferase (AST), alanine aminotransferase (ALT), and gamma-glutamyltransferase (GGT). Ultrasonographic findings of the gallbladder (size, shape, wall, content) and appearance of the biliary tract and the surrounding cranial intraabdominal organs were also evaluated. Percutaneous ultrasound-guided cholecystocentesis was performed easily during the study, and dogs tolerated well the procedure performed without anaesthesia. All laboratory parameters of the blood remained within normal limits throughout the study. However, some follow-up values, i.e. PCV, TPP, APTT and ALT, demonstrated statistically significant differences when compared to baseline measurements, which might reflect the effect of 24-hour fasting before the experiment, as well as day-to-day metabolic fluctuations due to feeding and water supply during the study. There were no visible signs of bleeding from the liver, bile leakage from the gallbladder or accumulation of free peritoneal fluid during repeated ultrasonographic examinations. Percutaneous ultrasound-guided cholecystocentesis seems to be an important diagnostic procedure in canine gallbladder diseases and can be used safely and easily to gain gallbladder bile for diagnosis of bacterial cholecystitis or for investigating hepatobiliary function in the dog.