First detection of Cupriavidus gilardii in a bovine neonatal diarrhea outbreak.

BACKGROUND: Cupriavidus gilardii is an aerobic, gram-negative, motile, glucose-nonfermenting bacillus, first described in 1999. Typically, it exhibits low pathogenicity in humans, causing opportunistic infections primarily in individuals with compromised immune systems. This bacterium has been also found in various environmental sources such as plants and contaminated soils. Notably, there have been no documented cases of C. gilardii infections in animals. CASE PRESENTATION: This case report outlines a bovine neonatal diarrhea outbreak that occurred in Northern Greece, during which C. gilardii was isolated. Faecal samples from 5-day-old calves were collected and transported to the laboratory for further examination. Bacterial culture and next generation sequencing techniques were employed to confirm the presence of this bacterium in the samples. Following the isolation and identification of C. gilardii from the samples, an autogenous vaccine was produced and administered to the cows within the farm. Subsequent to vaccination, a progressive reduction in calf diarrhea and deaths was observed, leading to their eventual complete resolution. To the best of our knowledge, this represents the first documentation of C. gilardii isolation from cases of bovine neonatal diarrhea. CONCLUSION: This case report presents the first isolation case of C. gilardii from animal samples and more specifically from calf faecal samples. It represents an important observation, providing evidence that this opportunistic human pathogen could contribute to clinical symptoms in animals.

The Effect of Iloprost in the Healing of Colonic Anastomosis in Rats under Chemotherapy with Irinotecan.

PURPOSE: We have investigated the possible positive effect of iloprost in the healing of colonic anastomosis, in rats under intraperitoneal chemotherapy with irinotecan. METHOD: Forty male Wistar rats have been divided into four groups. They underwent a partial colectomy and a termino-terminal anastomosis. They were administered, intraperitoneally, saline (group 1), irinotecan (group 2), iloprost (group 3), or irinotecan and iloprost (Group 4). After the sacrifice of the animals what followed was an autopsy, a macroscopic examination and the measurement of the anastomotic rupture pressure. A portion of the anastomosis was sent over for histological examination and determination of hydroxyproline levels. Results: The frequency of the leakage from the anastomosis was considered as significantly increased in group 2 compared with the other groups. In group 2, a significantly greater degree of adhesions, compared to all the remaining groups, was observed. The bursting pressure of the anastomosis was significantly lower in group 2, as compared with all the remaining groups, and significantly increased in the group 4 compared with group 2. Leukocytosis, fibroblasts, the neocollagen and the levels of hydroxyproline in group 4 showed significantly increased values, compared with group 2. The angiogenesis was significantly increased in groups 3 and 4 compared with group 2. Conclusions: Intraperitoneal administration of iloprost after colectomy, termino-terminal anastomosis and intraperitoneal administration of irinotecan promotes the healing process of the colon anastomoses as it competes the inhibitory effect of irinotecan.

Immunohistochemical Analysis of IL-19 and IL-24 Expression in Inflammatory Bowel Disease (IBD) Patients: Results From a Single Center Retrospective Study.

Background IL-19 and IL-24 induce proinflammatory cytokine production through the Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway. The primary objective of this study was to investigate any changes in IL-19 and IL-24 expression between inflammatory bowel disease (IBD) patients and healthy controls, as well as before and after the initiation of biologics. The secondary objective was to investigate any relation between their expression and disease phenotype and activity.  Methods IL-19 and IL-24 expression was measured in intestinal tissue samples from 121 patients with moderate to severe IBD versus healthy controls using immunohistochemistry. Their expression was then measured 12 months after treatment on the patient group treated with biologics. The disease activity was measured before and after treatment using the Harvey Bradshaw Index (HBI) for Crohn's disease (CD) patients and the Mayo Score (MS) for ulcerative colitis (UC) patients. Data were analyzed using SPSS (IBM Inc., Armonk, New York).  Results IL-19 expression was raised in the IBD group versus healthy controls. In the CD group, the IL-19 expression was related with the disease activity score post-biologic treatment. IL-24 was also highly expressed in patients with active UC and CD and was increased post-treatment. Its expression in UC was statistically related with the MS. Conclusions IL-24 and IL-19 are key factors in IBD-related intestinal inflammation and this is one of the few human studies to suggest that. An immunosuppressive role of IL-24 was demonstrated in the UC group. A future use as biomarkers of disease activity and response to treatment might be feasible.

Expression of IL-21 and IL-33 in Intestinal Mucosa of Inflammatory Bowel Disease: An Immunohistochemical Study.

Interleukins are considered to be potential therapeutic targets that can alter the prognosis and disease progression of IBD. IL-21 has proven to be involved in effector Th1, Th2 and Th17 responses. Similarly, IL-33, a newly identified cytokine, has been shown to control the Th1 effector response and the action of the colonic Tregs in animal models of colitis and patients with IBD. In this retrospective study, we have studied the expression of these interleukins, using immunohistochemistry, in 121 patients with moderate to severe IBD before and after treatment with biologics. The results were statistically processed using SPSSTM. Increased IL-21 expression was found in the UC and CD groups versus the controls. The IL-33 expression was found to be increased in the post-treatment UC and CD groups, suggesting a protective role of this interleukin against bowel inflammation. The IL-33 expression post-treatment was reversely correlated with the activity index score in CD patients, suggesting a better response to treatment in patients with higher IL-33 mucosa levels. This is the first immunohistochemical study of the expression of those interleukins in bowel mucosa before and after treatment with biologics. These data support a possibly promising future use of these interleukins as biomarkers of severe disease and response to treatment and as potential therapeutic targets for novel monoclonal antibodies.

Characterization of Extended Spectrum Cephalosporin-Resistant Escherichia coli Strains Isolated from Raw Poultry Carcasses in Catering Services in Northern Greece.

Antimicrobial resistance is considered a topic of utmost interest under the concept of "One Health", having severe implications in both human and veterinary medicine. Among the antibiotic-resistant bacteria, gram-negative bacteria, especially those belonging to the order of Enterobacterales (such as Escherichia coli), hold a prominent position in terms of both virulence and possessing/disseminating antimicrobial resistance (AMR) traits. The aim of this study was to examine the presence of extended-spectrum ß-lactamase producing E. coli isolates in raw poultry carcasses collected from a university club. Five hundred raw poultry skin samples were collected from the Aristotle University of Thessaloniki (AUTh) club in Thessaloniki, Greece. A total of 64% of the samples were positive for the presence of extended-spectrum ß-lactamase (ESBL)-producing E. coli. The isolates were further examined for their susceptibility to selected antibiotics by the disc diffusion method and were characterized as true ESBL, as producing class C cephalosporinases (AmpC) or "of unknown etiology" by the combination disc test. The 86 of the 120 isolates (71.67%) were classified as true ESBL, 24 (20.00%) as AmpC, and 10 (8.33%) as "of unknown etiology". The isolates were screened for the occurrence of ß-lactamase genes (blaTEM, blaCTX-M, blaSHV, and blaOXA). Thirty-six isolates (32 ESBL- and 4 AmpC-phenotype) harbored both blaTEM and blaCTX-M genes, twenty-two isolates (among which 19 ESBL-phenotype and 2 AmpC-phenotype) harbored blaCTX-M only, whereas twenty-six (14 ESBL- and 12 AmpC-phenotype) isolates harbored blaTEM alone. No isolate harboring blaSHV or blaOXA was detected. The results demonstrate the existence of E. coli isolates producing extended-spectrum ß-lactamases in poultry carcasses from Greece, pausing a risk for antibiotic resistance transfer to humans.

Adrenal glands of mice and rats: A comparative morphometric study.

Mice and rats are among the most used laboratory animals. They share numerous similarities along with differences, some yet unexplored. One of them is the morphometry of their adrenal glands, whose characteristics may be related to differences in energy management, immune response, drug metabolism, behaviour and temperament. The present study tries to fill this knowledge gap with the evaluation and comparison of adrenal gland anatomical/morphometric parameters of mice and rats. In groups of 10 (n = 10) adult, male and female BALB/c mice and Wistar rats, one in every 20 sections transverse to the longitudinal axis of the gland was used for measuring entire gland area, capsule, entire cortex, cortex zones and medulla with the aid of an image analysis system and subjected to statistical analysis. Quotients of the individual areas were calculated and comparison between the resulting ratios was performed. Gland length and volume were also calculated. Statistically significant differences were revealed between the rat female and male cortex area, rat and mouse medulla/cortex, medulla/gland, zona glomerulosa/cortex and cortex/gland ratios, male and female rats' medulla/cortex, medulla/gland, capsule/gland, zona glomerulosa/cortex, zona reticularis/cortex and zona glomerulosa/zona fasciculata ratios, length and volume. The correlation evaluation revealed that in male rats and in female mice the larger medulla area was accompanied by a larger cortex area and vice versa. In general, a larger cortex area was accompanied by larger areas of cortex zones. The collected data and the revealed differences can possibly contribute to the understanding of the physiology of the two species.

Socioeconomic Appraisal of an Early Prevention System against Toxic Conditions in Mussel Aquaculture.

This paper examines the financial viability and potential socioeconomic effects of introducing and operating an automated, remote-controlled management system for mussel farms which uses probes of temperature, dissolved oxygen, and conductivity associated with prediction software to demonstrate the potential need for mussel movement between marine areas. This system provides an early warning to farmers regarding the presence of toxins in aquatic ecosystems, thus contributing to saving mussel production and avoidikng significant economic losses. The analysis combines two established methodological tools in agricultural economics (linear programming and cost-benefit analysis) and provides estimates of the Net Present Value of the investment under two scenarios-one reflecting the existing situation and one a possible future situation where the mussel production system is expanded. The results of the analysis reveal the mid- and long-term effects of using the automated system, both of which demonstrate that the system is economically viable even if it contributes to saving mussel production from toxicity occurrence for only one year during its period of operation. The annual gross margin in the first scenario was €386,069 but almost tripled in the second scenario (€1,154,649). In addition, the future development and expansion of the mussel sector will likely be based on larger farms with an entrepreneurial and exporting orientation where risk mitigation systems, such as the one appraised in this paper, can play an important role.

Effect of Glyceryl Monoolein Addition on the Foaming Properties and Stability of Whipped Oleogels.

Medium Chain Triglyceride (MCT) oil was successfully combined with Glyceryl Monostearate (GMS) and Glyceryl Monoolein (GMO) to form oleogels that were subsequently whipped to form stable oleofoams. The co-crystallization of GMS and GMO at a ratio of 20:1, 20:2.5, and 20:5 within MCT oil was studied through Differential Scanning Calorimetry (DSC), X-ray Diffraction analysis (XRD), rheological analysis, Fluorescence Recovery after Photobleaching (FRAP), Fourier Transform Infrared Spectroscopy (FTIR), and polarized microscopy. The addition of 5% GMO resulted in the production of more stable oleogels in terms of crystal structure and higher peak melting point, rendering this formulation suitable for pharmaceutical applications that are intended to be used internally and those that require stability at temperatures close to 40 °C. All formulations were whipped to form oleofoams that were evaluated for their storage stability for prolonged period at different temperatures. The results show that oleofoams containing 5% MGO retained their foam characteristics even after 3 months of storage under different temperature conditions.

Effects of Dietary Lysolecithin Supplementation during Late Gestation and Lactation on Sow Reproductive Performance, Sow Blood Metabolic Parameters and Piglet Performance.

The objective of the present study was to evaluate the effects of dietary supplementation of lysolecithin in sows' diets during the last three weeks of the gestation period and throughout the lactation period on performance and metabolic parameters. In total 60 sows were allocated to two treatments: (a) CG (control group): the sows were fed commercially control diets; (b) LLG (lysolecithin group): the sows were fed the control diets supplemented with 750 g/t of feed supplemented with lysolecithin (Lysoforte Booster DryTM, Kemin Europa N.V., Herentals, Belgium). Backfat was lower in LLG than CG at end of gestation and at weaning (p = 0.030 and 0.044, respectively), while the CG sows mobilized more backfat between day 14 to weaning (p = 0.006). Litter weight at weaning was higher in the LLG (p = 0.027). Fasted glucose levels at day 14 of lactation tended to be lower in LLG compared to CG (p = 0.074). Urea concentrations were higher in LLG than CG at day 14 (p = 0.002). Lysolecithin supplemented sows compared to the control mobilized less tissue during lactation to support lactation demands. In conclusion, lysolecithin supplementation in sows resulted in improved litter weight at weaning without an excessive catabolism of backfat tissue, most probably due to an efficient nutrient utilization, which warrants further investigation.

Cataract Surgery during the COVID-19 Pandemic: Insights from a Greek Tertiary Hospital.

BACKGROUND: COVID-19 has affected everyday clinical practice, having an impact on the quality of healthcare provided, even in eye clinic departments. The aim of this study is to evaluate the consequences of this worldwide pandemic on cataract surgery in a Greek tertiary university hospital. METHODS: A total of 805 patients were included in this study. The number of cataract surgeries (CS), the type, the unilateral or bilateral appearance as well as the stage of cataract were recorded for the months between January and June 2019 (pre-COVID period) and compared with the same period in 2021 (during the pandemic outbreak) in the Department of Ophthalmology of Thessaloniki General Hospital G. Papanikolaou. RESULTS: A significant reduction in the number of CS as well as a significant increase in advanced and/or bilateral cataracts in 2021 compared to the pre-COVID period were observed. CONCLUSION: The COVID-19 pandemic has affected equally the value of ophthalmic interventions as well as the patients' quality of life, being a powerful reminder of the significant physical and psychological benefits of CS, especially for older adults and patients with comorbidities.

Phenotypic and Molecular Patterns of Resistance among Campylobacter coli and Campylobacter jejuni Isolates, from Pig Farms.

The purpose of this research was to characterize the antibiotic resistance patterns of Campylobacter spp. isolated from commercial farrow to finish farms in Greece, and analyze the relevant molecular resistance mechanisms among the resistant Campylobacter isolates. Susceptibility testing to five different classes of antibiotics was performed in 100 C. coli and 100 C. jejuni, previously isolated and identified. All isolates were found susceptible to meropenem. Very high rates of resistance were recorded for tetracyclines (84.5%), medium rates of resistance were recorded regarding quinolones (23%), and low and very low rates of resistance were identified for macrolides such as erythromycin and aminoglycosides (12% and 4%, respectively). Only 12.5% of the Campylobacter isolates displayed MDR. Regarding the molecular mechanisms of resistance, all ciprofloxacin resistant isolates hosted the mutant type Thr-86-Ile region of the quinolone resistance-determining region (QRDR) of the gyrA gene. In all erythromycin resistant isolates, the transitional mutations A2075G and A2074C in the 23S rRNA gene were only amplified. Molecular screening of tetracycline resistance genes indicated that the vast majority of Campylobacter isolates (92.3%) were positive for the tet(O) gene. In summary, these findings and especially the very high and medium rates of resistance for tetracyclines and fluroquinolones, respectively recommend that a continuous monitoring of Campylobacter isolates susceptibility in combination with the proper use of antimicrobials in livestock production is of great importance for public health.

Histological Effects of Intravitreal Injection of Antifungal Agents in New Zealand White Rabbits: An Electron Microscopic and Immunohistochemical Study.

Fungal endophthalmitis is a serious and vision-threatening infection which requires an immediate and effective treatment approach. Our research aims to elucidate the histological effects of the intravitreal injection of the maximum safe dosage of voriconazole and micafungin on retina. Six albino New Zealand White Rabbits were used. In experimental animals, a solution of voriconazole (Group V) or micafungin (Group M) was intravitreally injected in the right eye, while in control animals, balanced salt solution was intravitreally injected in the left eye (Group C). Euthanasia was performed ten days post injection and the retina was removed and prepared for histological examination with a light and electron microscope. Eosin-hematoxylin staining did not reveal any pathological changes in any of the samples examined. The immunohistochemical staining for Tumor Necrosis Factor alpha (TNF-a) marker was detected as negative in all samples, while Interleukin 6 (IL-6) marker was detected as mild only in the group injected with voriconazole. Electron microscopy revealed several ultrastructural alterations in retinal layers in both groups of experimental animals. Histological retinal lesions, revealed with electron microscopy in the present investigation, raises the question of the safe usage of these antifungal agents in the treatment of fungal intraocular infections in the future.

Application and validation of a LC/fluorescence method for the determination of amoxicillin in sheep serum and tissue cage fluid.

A LC method with fluorescence detection after pre-column mercury dichloride derivation was developed and validated for the quantitative determination of amoxicillin in sheep blood serum and tissue cage fluid at levels down to 100 and 200ng/mL, respectively. Spiked blood serum and tissue cage fluid samples were deproteinized, derivatized with mercury dichloride and extracted prior to reversed phase LC analysis with fluorescence spectrophotometric detection at an excitation wavelength of 355nm and an emission wavelength of 435nm. Separation was carried out on a C(18) column with a mobile phase consisting of phosphate buffer, octanesulphonate sodium (OCT), and acetronitrile. A regression model using 1/concentration weighting was found the most appropriate for quantification. The intra-day precision for serum was 1.65-8.74% and for tissue cage fluid was 2.48-6.27%. The inter-day precision for serum was 0.39-3.57% and for tissue cage fluid was 0.44-2.54%. The overall precision over 3 days for blood serum using of 108 replicates was 1.70-9.44% and for tissue cage fluid using of 54 replicates was 2.51-6.76%. Studies of amoxicillin stability in blood serum and tissue cage fluid indicated that amoxicillin was stable after 4 weeks storage at -85 degrees C. The method was successfully applied for the determination of amoxicillin in blood serum and tissue cage fluid samples collected from rams after intravenous administration.

Pharmacodynamic interactions of amikacin with selected ß-lactams and fluoroquinolones against canine Escherichia coli isolates.

Knowledge of in vitro antimicrobial interactions can serve as a guide for clinical application of combination antimicrobial regimens. The aim of the present study was to determine the pharmacodynamic interactions of amikacin with either amoxicillin/clavulanic acid, ceftazidime, enrofloxacin or marbofloxacin against clinical canine Escherichia coli isolates. Bactericidal activity of individual antimicrobials was assessed by use of static kill curves. Interactions between amikacin and each of the ß-lactams or fluoroquinolones were subsequently analyzed by employing the fractional maximal effect method. Amikacin, compared with all other agents, displayed the most rapid and extensive bacterial killing, the lowest level (with respect to MIC) at which half the maximal effect was observed and the most linear concentration-effect relationship. The combinations of amikacin with amoxicillin/clavulanic acid or ceftazidime were completely synergistic in four and three out of the five investigated isolates, respectively, with additivity being sporadically observed. On the other hand, the combinations of amikacin with enrofloxacin or marbofloxacin yielded a mosaic of interaction types with no discernible pattern or differentiation between fluoroquinolone-susceptible and resistant isolates; synergy was only infrequently observed, mainly at increased fluoroquinolone concentrations. In conclusion, the combinations of amikacin with the two ß-lactams were found to be more promising, in terms of synergy achievement, compared with the respective combinations with the two fluoroquinolones.

Prevalence and mechanisms of resistance to fluoroquinolones in Pseudomonas aeruginosa and Escherichia coli isolates recovered from dogs suffering from otitis in Greece.

The aim of this study was to investigate the prevalence and the implicated mechanisms of resistance against selected veterinary fluoroquinolones (enrofloxacin, marbofloxacin and pradofloxacin) among 101 Pseudomonas aeruginosa (n=75) and Escherichia coli (n=26) isolates collected from dogs suffering from otitis. Resistance ranged from 32.0% to 48.0% with differences not being considered statistically significant among the three agents or between the two bacterial species. However, individual MICs of pradofloxacin, the latest veterinary fluoroquinolone, were significantly lower than those of enrofloxacin, the oldest one, indicating an increased in vitro potency of the former antimicrobial. Pradofloxacin MIC90 was, additionally, the lowest (8µg/ml), in E. coli, or among the lowest (8µg/ml), in P. aeruginosa isolates. Resistance was in most cases associated with topoisomerase substitutions, with patterns GyrA:V73G in P. aeruginosa and GyrA:S83L+D87N/ParC:S58I+A86V in E. coli being reported for the first time in small animal isolates. Only 6.7% and 15.4% of P. aeruginosa and E. coli otitis isolates, respectively, carried plasmid-mediated quinolone resistance (PMQR) genes, which, moreover, contributed minimally to resistance. Efflux pump activity was additionally detected in resistant E. coli isolates, even those lacking topoisomerase substitutions or PMQR genes. The emergence of resistance in the canine otitis isolates seemed to be associated with previous, prolonged systemic fluoroquinolone administration. In any case, antimicrobial susceptibility testing should guide the selection of systemic FQs for the treatment of canine otitis.

Clindamycin bioavailability and pharmacokinetics following oral administration of clindamycin hydrochloride capsules in dogs.

Oral bioavailability and pharmacokinetic behaviour of clindamycin in dogs was investigated following intravenous (IV) and oral (capsules) administration of clindamycin hydrochloride, at the dose of 11 mg/kg BW. The absorption after oral administration was fast, with a mean absorption time (MAT) of 0.87+/-0.40 h, and bioavailability was 72.55+/-9.86%. Total clearance (CL) of clindamycin was low, after both IV and oral administration (0.503+/-0.095 vs. 0.458+/-0.087 L/h/kg). Volume of distribution at steady-state (IV) was 2.48+/-0.48 L/kg, indicating a wide distribution of clindamycin in body fluids and tissues. Elimination half-lives were similar for both routes of administration (4.37+/-1.20 h for IV, vs. 4.37+/-0.73 h for oral). Serum clindamycin concentrations following administration of capsules remained above the MICs of very susceptible microorganisms (0.04-0.5 microg/mL) for 12 or 10 h, respectively. Time above the mean inhibitory concentration (MIC) is considered as the index predicting the efficacy of clindamycin (T(>MIC) must be at least 40-50% of the dosing interval), so a once-daily oral administration of 11 mg/kg BW of clindamycin can be considered therapeutically effective. For less susceptible bacteria (with MICs of 0.5-2 microg/mL) the same dose should be given but twice daily.

Reversed-phase liquid chromatographic method with fluorescence detection for the simultaneous determination of albendazole sulphoxide, albendazole sulphone and albendazole 2-aminosulphone in sheep plasma.

A rapid and sensitive HPLC method for the simultaneous quantification of albendazole sulphoxide (ABZ-SO), albendazole sulphone (ABZ-SO2) and albendazole 2-aminosulphone (ABZ-SO2NH2) in sheep blood plasma has been developed. Plasma samples were extracted with ethyl acetate under alkaline conditions. Separation was achieved on a C18 reversed-phase analytical column, in the presence of positively- (tetra-n-butylammonium hydrogen sulphate) and negatively-charged (octanesulphonate sodium) pairing ions, while detection was performed fluorometrically. Excitation and emission wavelengths were 290 and 320 nm, respectively. Limits of quantification were defined at 39 ng/ml for ABZ-SO, 4.95 ng/ml for ABZ-SO2 and 4 ng/ml for ABZ-SO2NH2. Accuracy data, in terms of recovery efficiency showed overall values (+/- S.E.M.) of 85.6 +/- 1.0% for ABZ-SO, 100.0 +/- 1.0% for ABZ-SO2 and 89.1 +/- 0.6% for ABZ-SO2NH2. The method was successfully applied to quantitatively determine the three albendazole metabolites in plasma samples collected from sheep that had been orally administered albendazole.

Peripheral distribution of amoxicillin in sheep and influence of local inflammation.

The pharmacokinetics of amoxicillin (AMX) in blood serum (SBS) and tissue cage fluid (TCF) was studied in sheep. Four tissue cages, prepared from silicone rubber tubing, were subcutaneously inserted in the neck area (two on each side) of the experimental animals and AMX was administered both intravenously (IV) and intramuscularly (IM) at the dose rate of 15mg/kg bodyweight. The impact of local inflammation on AMX distribution in TCF was studied after intra-cavity injection of a lambda carrageenan solution in one of the two tissue cages used after each administration. In contrast to the three-compartment AMX disposition after IV injection, two-compartment, absorption-limited pharmacokinetics was observed after IM administration. Non-inflamed and inflamed TCF data revealed, in all cases, the attainment of low, but prolonged concentrations and absence of an inflammation-induced effect on AMX penetration into and elimination from TCF.