Supplementation of vitamin E as an addition to a commercial renal diet does not prolong survival of cats with chronic kidney disease.

BACKGROUND: The aim of this double-blind, placebo-controlled study was to investigate the effect of vitamin E supplementation as an addition to a commercial renal diet on survival time of cats with different stages of chronic kidney disease (CKD). In addition, we were interested whether vitamin E supplementation affects selected oxidative stress and clinical parameters. Thirty-four cats with CKD and 38 healthy cats were included in the study. Cats with CKD were classified according to the IRIS Guidelines; seven in IRIS stage 1, 15 in IRIS stage 2, five in IRIS stage 3 and seven in IRIS stage 4. Cats with CKD were treated according to IRIS Guidelines. Cats with CKD were randomly assigned to receive vitamin E (100 IU/cat/day) or placebo (mineral oil) for 24 weeks in addition to standard therapy. Plasma malondialdehyde (MDA) and protein carbonyl (PC) concentrations, DNA damage of peripheral lymphocytes and plasma vitamin E concentrations were measured at baseline and four, eight, 16 and 24 weeks thereafter. Routine laboratory analyses and assessment of clinical signs were performed at each visit. RESULTS: Vitamin E supplementation had no effect on the survival time and did not reduce the severity of clinical signs. Before vitamin E supplementation, no significant differences in vitamin E, MDA and PC concentrations were found between healthy and CKD cats. However, plasma MDA concentration was statistically significantly higher (p = 0.043) in cats with early CKD (IRIS stages 1 and 2) than in cats with advanced CKD (IRIS stages 3 and 4). Additionally, DNA damage was statistically significantly higher in healthy cats (p ≤ 0.001) than in CKD cats. Plasma vitamin E concentrations increased statistically significantly in the vitamin E group compared to the placebo group four (p = 0.013) and eight (p = 0.017) weeks after the start of vitamin E supplementation. During the study and after 24 weeks of vitamin E supplementation, plasma MDA and PC concentrations and DNA damage remained similar to pre-supplementation levels in both the placebo and vitamin E groups. CONCLUSIONS: Vitamin E supplementation as an addition to standard therapy does not prolong survival in feline CKD.

Incidence of Aflatoxins and Ochratoxin A in Wheat and Corn from Albania.

In this study, aflatoxins (AFs) and ochratoxin A (OTA) were analyzed in grains, specifically wheat and corn, from Albania. To summarize, 71 wheat and 45 corn samples from different growing areas were collected. The multi-toxin analytical procedure involved sample extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The incidence of AF was 18% in the analyzed wheat and 71% in the corn samples. The concentration of AFs was much higher in the corn samples than in the wheat samples. The maximum permitted levels for aflatoxin B1 (AFB1) and total AFs were not exceeded in the wheat samples, while they were exceeded in 36% of the corn samples. In the wheat samples, the AFB1 concentration varied between 0.2 and 0.4 µg kg-1. However, the highest concentrations in the corn samples were 2057, 2944, and 3550 µg kg-1. OTA was present in only three corn samples and one wheat sample. However, all contaminated samples exceeded the maximum permitted levels. This report reveals the presence of AFs and OTA in grain commodities, specifically wheat and corn, grown in Albania.

Effects of Coenzyme Q10 Supplementation on Oxidative Stress Markers, Inflammatory Markers, Lymphocyte Subpopulations, and Clinical Status in Dogs with Myxomatous Mitral Valve Disease.

Scarce data exist on the effects of coenzyme Q10 (CoQ10) supplementation in dogs with myxomatous mitral valve disease (MMVD). The purpose of this study was to investigate the effect of CoQ10 supplementation on oxidative stress markers (glutathione peroxidase, F2-isoprostanes), markers of inflammation (tumor necrosis factor-α, TNF soluble receptor II, leucocytes, and their subtypes), lymphocyte subpopulations (T helper and cytotoxic T lymphocytes, including activated T lymphocytes, and B lymphocytes), and echocardiographic and clinical parameters in dogs with MMVD. In this randomized, controlled, double-blind, longitudinal study, 43 MMVD dogs in stages ACVIM (American College of Veterinary Internal Medicine classification) B2 and ACVIM C and D (congestive heart failure (CHF)) received water-soluble coenzyme Q10 (100 mg twice daily) or placebo for 3 months, and 12 non-supplemented healthy dogs served as controls. All parameters were measured before and after supplementation in MMVD dogs and once in healthy dogs. CoQ10 supplementation had a positive impact on neutrophil percentage, lymphocyte percentage, and lymphocyte concentration in our cohort of dogs with CHF (ACVIM C and D). Conclusion: CoQ10 as an oral supplement may have benefits in terms of decreasing inflammation in dogs with MMVD and CHF.

Occurrence of Alternaria and Other Toxins in Cereal Grains Intended for Animal Feeding Collected in Slovenia: A Three-Year Study.

In recent years, the less-studied Alternaria mycotoxins have attracted increasing interest due to the lack of survey data and their ability to cause toxic effects in animals and humans. To fill the gap, the aim of this three-year survey was to investigate the presence and co-occurrence of Alternaria and other mycotoxins in a total of 433 cereal grain samples from Slovenian farms and agricultural cooperatives from 2014 to 2016. Using the multi-mycotoxin method, 14 mycotoxins were determined. In 53% of 433 analysed samples, contamination with at least one mycotoxin was found. Deoxynivalenol (DON) and tenuazonic acid (TeA) were present in 32% and 26% of cereal grain samples, respectively, whereas alternariol (AOH), tentoxin (TEN), alternariol monomethyl ether (AME), 3- and 15-acetyldeoxynivalenol (3- and 15-AcDON), and zearalenone (ZEN) were present in fewer than 15% of the samples. Ochratoxin A (OTA) was found in one rye sample, while diacetoxyscirpenol (DAS), HT-2 and T-2 toxin, and fumonisins B1 and B2 (FB1 and FB2) were not detected. The highest maximum and median concentrations of Alternaria toxins were determined in spelt in 2016 (TeA, 2277 µg/kg and 203 µg/kg, respectively), and those of Fusarium toxins in wheat in 2015 (DON, 4082 µg/kg and 387 µg/kg, respectively). The co-occurrence of two or more mycotoxins was found in 43% of the positive samples. The correlations between Alternaria toxins were very weak but statistically significant (r: 0.15-0.17, p: 0.0042-0.0165). A well-known correlation between Fusarium toxins DON and ZEN was weak and highly significant (r = 0.28, p < 0.0001).

Randomized, double-blinded, controlled trial of the effects of coenzyme Q10 supplementation on plasma coenzyme Q10 concentration in dogs with myxomatous mitral valve disease.

OBJECTIVE: To determine the dose of coenzyme Q10 (CoQ10) needed to achieve at least a 3-fold increase in plasma CoQ10 concentration in dogs with myxomatous mitral valve disease (MMVD) and congestive heart failure (CHF). ANIMALS: 18 dogs with CHF due to MMVD and 12 healthy dogs. PROCEDURES: In a randomized, double-blinded, controlled trial, dogs with MMVD were given 50 or 100 mg of water-soluble CoQ10 (ubiquinone; total daily dose, 100 mg [n = 5] or 200 mg [6]) or a placebo (7), PO, twice a day for 2 weeks in addition to regular cardiac treatment. Plasma CoQ10 concentration was measured in dogs with MMVD before (baseline) and at various time points after supplementation began and in healthy dogs once. Concentrations were compared among and within groups. RESULTS: No significant difference in median baseline plasma CoQ10 concentration was detected between healthy dogs and dogs with MMVD. Fold increases in plasma CoQ10 concentrations ranged from 1.7 to 4.7 and 3.2 to 6.8 for individual dogs in the 100-mg and 200-mg groups, respectively. The change in plasma CoQ10 concentration after supplementation began was significantly higher than in the placebo group at 4 hours and 1 and 2 weeks for dogs in the 200-mg group and at 1 and 2 weeks for dogs in the 100-mg group. CONCLUSIONS AND CLINICAL RELEVANCE: A daily CoQ10 dose of 200 mg was sufficient to achieve at least a 3-fold increase in plasma CoQ10 concentration and may be used in CoQ10 supplementation studies involving dogs with CHF due to MMVD.

Entomological Surveillance and Cantharidin Concentrations in Mylabris variabilis and Epicauta rufidorsum Blister Beetles in Slovenia.

True blister beetles (genus Epicauta, family Meloidae) produce cantharidin, which can cause toxicosis in humans and animals. Some recent reports suggest that poisoning by the blister beetle has occurred in the Mediterranean part of Slovenia, which has never been reported before. Drought and modern harvesting techniques are thought to increase the likelihood of blister beetle forage contamination and cantharidin intoxication in animals. A survey of fields associated with blister beetle contamination was conducted and the Meloid species present were identified. Entomological surveillance was conducted for Mylabris variabilis and Epicauta rufidorsum. Cantharidin concentrations were also measured in both blister beetle species. Cantharidin concentration in Mylabris variabilis (n = 17) ranged from 0.038 to 0.354 µg/mg (mean 0.151 µg/mg). Cantharidin concentration in Epicauta rufidorsum (n = 36) ranged from 0.055 to 0.341 µg/mg (mean 0.142 µg/mg). Both species exhibited variable concentrations of cantharidin that could not be associated with their biology, sex, age, size, and/or reproductive status. Epicauta rufidorsum have never previously been studied as a possible source of forage contamination, nor have cantharidin concentrations been determined in this species. It is the most likely source of forage contamination due to its abundance in the investigated fields, its swarming activity, and its tendency to reside in the green parts of plants immediately after cutting. Delaying the simultaneous processing and storage of forage after cutting would reduce the likelihood of forage contamination by blister beetles, as they can then retreat to the ground or fly away.

Ergot and Ergot Alkaloids in Cereal Grains Intended for Animal Feeding Collected in Slovenia: Occurrence, Pattern and Correlations.

This four-year study reports the occurrence of ergot alkaloids (EAs) in cereals intended for animal feeding collected in Slovenia. A total of 517 samples of cereals were analysed using liquid chromatography-tandem mass spectrometry for the presence of EAs. The sample set included wheat, rye, triticale, oat, spelt and barley. The study revealed that 17% of the analysed cereal samples were contaminated with at least one ergot alkaloid. EAs have two epimeric forms: -ine and -inine. The incidence rates of the -ine and -inine forms in the analysed samples were 16% and 15%, respectively. The highest contamination rates were observed in rye (54%), oat (50%) and spelt (30%), where the highest mean concentrations of total EAs were also determined (502 µg/kg, 594 µg/kg and 715 µg/kg, respectively). However, the highest concentrations of total EAs were found in wheat and rye (4217 µg/kg and 4114 µg/kg, respectively). The predominant EAs were ergometrine, ergosine and ergocristinine. The occurrence of six or more ergot alkaloids was observed in 49% of the positive samples. A weak correlation (p = 0.284) in the positive samples was found between the mass of sclerotia and the total concentrations of EAs using the Spearman correlation coefficient.

Incidence of Fusarium Mycotoxins in Wheat and Maize from Albania.

In this study, ten Fusarium toxins were analysed in wheat and maize commodities from Albania. In total, 71 samples of wheat and 45 samples of maize were collected from different producing regions. The analytical procedure consisted of a simple one-step sample extraction followed by the determination of toxins using liquid chromatography coupled with tandem mass spectrometry. Fusarium toxins were found in 23% of the analysed wheat samples and in 78% of maize samples. In maize samples, most often fumonisins B1 (FB1) and B2 (FB2) were found. They were present in 76% of samples. They were detected in all positive samples except in one with concentrations ranging from 59.9 to 16,970 µg/kg. The sum of FB1 and FB2 exceeded the EU maximum permitted level (4000 µg/kg) in 31% of maize samples. In wheat samples, the only detected Fusarium mycotoxin was deoxynivalenol (DON), present in 23% of samples. In one sample with the concentration of 1916 µg/kg, the EU maximum permitted level (1250 µg/kg) was exceeded. This is the first report on the presence of Fusarium toxins in wheat and maize grains cultivated in Albania.

Mycotoxin Decontamination Efficacy of Atmospheric Pressure Air Plasma.

Mycotoxins, the toxic secondary metabolites of mould species, are a growing global concern, rendering almost 25% of all food produced unfit for human or animal consumption, thus placing immense pressure on the food supply chain. Cold Atmospheric pressure Plasma (CAP) represents a promising, low-cost, and environmentally friendly means to degrade mycotoxins with negligible effect on the quality of food products. Despite this promise, the study of CAP-mediated mycotoxin degradation has been limited to a small subset of the vast number of mycotoxins that plague the food supply chain. This study explores the degradation of aflatoxins, trichothecenes, fumonisins, and zearalenone using CAP generated in ambient air. CAP treatment was found to reduce aflatoxins by 93%, trichothecenes by 90%, fumonisins by 93%, and zearalenone by 100% after 8 minutes exposure. To demonstrate the potential of CAP-mediated mycotoxin degradation against more conventional methods, its efficiency was compared against ultraviolet C (UVC) light irradiation. In all cases, CAP was found to be considerably more efficient than UVC, with aflatoxin G1 and zearalenone being completely degraded, levels that could not be achieved using UVC irradiation.

Mycotoxin Decontamination of Food: Cold Atmospheric Pressure Plasma versus "Classic" Decontamination.

Mycotoxins are secondary metabolites produced by several filamentous fungi, which frequently contaminate our food, and can result in human diseases affecting vital systems such as the nervous and immune systems. They can also trigger various forms of cancer. Intensive food production is contributing to incorrect handling, transport and storage of the food, resulting in increased levels of mycotoxin contamination. Mycotoxins are structurally very diverse molecules necessitating versatile food decontamination approaches, which are grouped into physical, chemical and biological techniques. In this review, a new and promising approach involving the use of cold atmospheric pressure plasma is considered, which may overcome multiple weaknesses associated with the classical methods. In addition to its mycotoxin destruction efficiency, cold atmospheric pressure plasma is cost effective, ecologically neutral and has a negligible effect on the quality of food products following treatment in comparison to classical methods.

Occurrence of ergot alkaloids in wheat from Albania.

The occurrence of ergot alkaloids in wheat harvested in Albania was investigated. A total of 71 samples of winter wheat collected in 2014 and 2015 were analysed for the 12 most important ergot alkaloids using liquid chromatography-tandem mass spectrometry. In the harvesting year 2014, 48.6% of samples were contaminated with ergot alkaloids, whereas in 2015 only 19.4% of samples were contaminated. In 2014, the concentrations of total ergot alkaloids ranged from 17.3 to 975.4 µg kg-1, and in 2015 they ranged from 10.3 to 390.5 µg kg-1. The samples contained from one to nine ergot alkaloids. The most frequent were ergometrine, ergosine and ergocristine, and the least frequent were ergocryptine, ergocryptinine and ergocorninine.

Determination of N-acylhomoserine lactones of Pseudomonas aeruginosa in clinical samples from dogs with otitis externa.

BACKGROUND: Bacterial intercellular communication, called quorum sensing, takes place via the production and collective response to signal molecules. In Gram-negative bacteria, like Pseudomonas aeruginosa, these signaling molecules are N-acylhomoserine lactones (AHLs). P. aeruginosa is a common cause of inflammation of the ear canal (otitis externa) in dogs. It employs quorum sensing to coordinate the expression of host tissue-damaging factors, which are largely responsible for its virulence. The treatment of P. aeruginosa-associated otitis is challenging due to a high intrinsic resistance of P. aeruginosa to several antibiotics. Attenuation of quorum sensing signals to inhibit bacterial virulence is a novel strategy for the treatment of resistant bacterial pathogens, including P. aeruginosa. Therefore, it is important to recognize and define quorum sensing signal molecules in clinical samples. To date, there are no reports on determination of AHLs in the veterinary clinical samples. The purpose of this study was to validate an analytical procedure for determination of the concentration of AHLs in the ear rinses from dogs with P. aeruginosa-associated otitis externa. Samples were obtained with rinsing the ear canals with physiological saline solution. For validation, samples from healthy dogs were spiked with none or different known amounts of the selected AHLs. With the validated procedure, AHLs were analyzed in the samples taken in weekly intervals from two dogs, receiving a standard treatment for P. aeruginosa-associated otitis externa. RESULTS: Validation proved that the procedure enables quantification of AHLs in non-clinical and clinical samples. In addition, a time dependent reduction of AHL concentration was detected for the treated dogs. CONCLUSIONS: Our results indicate that liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is superior in detecting AHLs compared to other chromatographic techniques. This is the first report on determination of AHLs in the clinical samples of veterinary importance. The analytical procedure described in this paper is capable of supporting novel antimicrobial strategies, which target quorum sensing.

Plasma and skin vitamin E concentrations in canine atopic dermatitis.

BACKGROUND: Altered homeostasis of vitamin E has been demonstrated in human atopic dermatitis. Data on plasma and skin vitamin E concentrations in canine atopic dermatitis (CAD) are not available. OBJECTIVE: To determine vitamin E concentrations in plasma and skin of atopic dogs. ANIMALS AND METHODS: Vitamin E concentrations in plasma and full-thickness skin biopsies of 15 atopic dogs were related to CAD extent and severity index (CADESI-03) scores and compared to the equivalent concentrations in 17 healthy dogs. Statistically significant differences of measured parameters between the two groups were determined by the nonparametric Mann Whitney U test and correlations between CADESI-03 scores and vitamin E concentrations were evaluated by the Spearman rank test. A value of P < 0.05 was considered significant. RESULTS: Plasma concentrations of vitamin E were significantly lower in atopic dogs than in healthy dogs, with median values of 29.8 and 52.9 µmol/L, respectively. Skin vitamin E values did not differ significantly between patients and healthy controls. The median concentration of skin vitamin E in atopic dogs was higher than that in healthy dogs. No significant correlations were found between CADESI-03 score and plasma vitamin E or skin vitamin E concentrations. CONCLUSIONS: Significantly lower plasma vitamin E concentrations in atopic dogs than in healthy controls indicate altered homeostasis of vitamin E in CAD. CLINICAL IMPORTANCE: Further investigation into vitamin E supplementation in CAD is warranted.

Detection of trace amounts of abamectin used as an antiparasitic agent in fallow deer tissues.

A sensitive and reliable method for the determination of trace amounts of abamectin in muscles, kidneys and fat tissue of fallow deer is presented. Abamectin was extracted from the tissues with acetonitrile and the extract was cleaned up on a C8 solid-phase extraction cartridge. Abamectin residue was derivatised with trifluoroacetic acid anhydride and 1-methylimidazole, and determined using reversed-phase high-performance liquid chromatography under isocratic conditions and fluorescence detection. The recoveries of the method were high and consistent, ranging from 78% to 90%. The limit of detection of the method was below 1 microg/kg when analysing muscle, kidney and fat tissue. Matrix-matched calibration was used in order to obtain accurate values and to avoid matrix interference.