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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.

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.

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.

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.

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.

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.

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.

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 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.

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.

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.

The aim of the study was to examine the possible involvement of cholecystokinin (by lorglumide) and cholinergic mechanisms (by atropine) in magnesium sulphate (MgSO(4))-induced gallbladder contraction of conscious dogs. The gallbladder (GB) volume was determined by ultrasonography. The optimal dose of 80 mg kg(-1)of MgSO(4)was determined from a MgSO(4)dose-response curve using doses of 10, 20,40, 80, 120 mg kg(-1). The largest dose of MgSO(4)was less effective than the optimal dose. Peak gallbladder contraction (32 per cent) was achieved at 30 minutes. Atropine (50 microg kg(-1)s.c.) or lorglumide (1 mg kg(-1)p.o.) fully prevented GB contraction. In conclusion, supraoptimal doses of MgSO(4)have a diminishing effect. The sustained contraction of the gallbladder in response to the optimal dose of MgSO(4)can be explained by an additive effect of the cholecystokinin release and a cholinergic trigger mechanism. Ultrasonography and MgSO(4)stimulation proved to be a valuable technique for examination of gallbladder motility.