β-Carotene from the Alga Dunaliella bardawil Decreases Gene Expression of Adipose Tissue Macrophage Recruitment Markers and Plasma Lipid Concentrations in Mice Fed a High-Fat Diet

Vitamin A and provitamin A carotenoids are involved in the regulation of adipose tissue metabolism and inflammation. We examined the effect of dietary supplementation using all-trans and 9-cis β-carotene-rich Dunaliella bardawil alga as the sole source of vitamin A on obesity-associated comorbidities and adipose tissue dysfunction in a diet-induced obesity mouse model. Three-week-old male mice (C57BL/6) were randomly allocated into two groups and fed a high-fat, vitamin A-deficient diet supplemented with either vitamin A (HFD) or β-carotene (BC) (HFD-BC). Vitamin A levels in the liver, WATs, and BAT of the HFD-BC group were 1.5–2.4-fold higher than of the HFD group. BC concentrations were 5–6-fold greater in BAT compared to WAT in the HFD-BC group. The eWAT mRNA levels of the Mcp-1 and Cd68 were 1.6- and 2.1-fold lower, respectively, and the plasma cholesterol and triglyceride concentrations were 30% and 28% lower in the HFD-BC group compared with the HFD group. Dietary BC can be the exclusive vitamin A source in mice fed a high-fat diet, as shown by the vitamin A concentration in the plasma and tissues. Feeding BC rather than vitamin A reduces adipose tissue macrophage recruitment markers and plasma lipid concentrations.     

Reproducibility of bovine neonatal pancytopenia (BNP) via the application of colostrum

A haemorrhagic diathesis has been observed in young calves since 2007 which is described as bovine neonatal pancytopenia (BNP) and presents a completely new disease. The objectives of our investigation were to test if BNP could be reproduced using colostrum of cows with a BNP history and pre-colostral calves from farms where BNP has not been observed. In the present experiment, 22 German Holstein calves from BNP-free farms were fed four to six hours after birth 2.5 l colostrum from cows which had been reported to have had at least one calf with BNP in the last lactation. We distinguished three different experimental groups according to the composition of the colostrum. In experimental group I, each of the six calves received colostrum of a single cow, in experimental group II all six calves received colostrum from the same cow and in experimental group III each of the ten calves received a colostrum mix from ten different cows. Clinical signs of BNP were observed in 50% of the calves in experimental group I, 67% of the calves in experimental group II and all calves in experimental group III. The lethality in the three experimental groups was significantly different with rates of 16.7%, 66.7% and 80%, respectively. Calves fed with a colostrum-mix in experimental group III had the highest lethality. Neither the farm nor the amount of the colostrum fed had a significant effect on the occurrence and course of BNP. The profiles for thrombocytes, leucocytes and erythrocytes significantly differed in dependence of the severity of BNP signs. Calves with non-lethal BNP showed thrombocytopenia with values below 100 G/l on the 1th to 3rd and the 7th to 11th day of life. In calves with lethal BNP, thrombocytes decreased under 50 G/l from day 5. In calves with non-lethal BNP, a decrease of the leucocytes under the threshold was present only for a short period of time. In calves with lethal BNP, leucocytes decreased in the first 5 days after birth continuously and increased on the 6th to the 8th day to normal values and then a rapid decrease occurred. Erythrocytes decreased under the normal threshold just in the last two days before the calves died or were euthanized. Thus, the present experiments showed that colostrum of cows with a BNP-history and vaccination with PregSure BVD from Pfizer caused lethal BNP. We can assume that the different reactions of the calves are due to immunogenetic reactions to colostral alloreactive antibodies. The reaction spectrum of calves depends on the presence of antigens which can react with these colostral antibodies. The experimental results can explain the different incidences of BNP within and among farms as well as between breeds.     

Assessment of motor pathways by magnetic stimulation in human and veterinary medicine

Magnetic stimulation is a non-invasive and painless technique for studying the motor pathways in medical neurology. A time-varying magnetic field induces an electrical field in conducting objects, such as nervous tissue. The technique can be applied to nerve roots and peripheral nerves or to the motor cortex of the brain in human and veterinary medicine. In this review, the basic principles, applications and risk factors of peripheral nerve and motor cortex stimulation in human and veterinary medicine are discussed.     

Methicillin-resistant Staphylococcus pseudintermedius among dogs admitted to a small animal hospital

The aim of this study was to determine the frequency of carriage of methicillin-resistant Staphylococcus pseudintermedius (MRSP) among dogs admitted to a small animal hospital during a 17-month period, to characterize these isolates and to initially screen for possible factors associated with MRSP carriage. Swabs were taken from the nose/pharynx and the perineum as well as from wounds and skin infections (if present) of 814 dogs before entering the small animal hospital. A questionnaire for background information was completed. The staphylococcal species and methicillin resistance were confirmed pheno- and genotypically. The identified MRSP isolates were characterized by SCCmec typing, testing for susceptibility to 25 antimicrobial agents and SmaI-directed pulsed-field gel electrophoresis. A first screening for possible risk factors for MRSP carriage was performed by means of unifactorial contingency tables and CART analysis. Sixty (7.4%) dogs were positive for MRSP. All MRSP isolates harboured a type II-III SCCmec cassette and showed extended resistance to antimicrobial agents. Fifteen different SmaI patterns were observed. The major factors that clustered with MRSP carriage were former hospitalization and antibiotic treatment within the last six months before sampling. This study showed that only a minor part of the sampled dogs carried multi-resistant MRSP isolates. The facts that prior hospitalization and/or antibiotic therapy are potential associated factors for MRSP carriage underline the necessity of a judicious use of antibiotics in small animal medicine.     

Comparison of bone marrow aspiration at the sternum and the tuber coxae in middle-aged horses

The objective of this study was to compare bone marrow (BM) aspirates from the sternum and the tuber coxae of middle-aged horses. Bone marrow was obtained from the sternum and both tubera coxae of 12 healthy, 13-year-old geldings. Two different puncture techniques were used for the tuber coxae. The 2 syringes used for sternal sampling were evaluated separately. The mononuclear cell (MNC) fraction of the BM was isolated and the mesenchymal stem cells (MSCs) were culture-expanded. At the sternum, BM aspiration was always possible. Bone marrow aspiration at the tuber coxae required straight and deep needle penetration combined with high negative pressure. With this technique a median sample amount of 11.0 mL with large individual variation was obtained. A median of 3.06 × 10(6) MNC/mL BM (1st syringe) and 2.46 × 10(6) MNC/mL BM (2nd syringe) was isolated from sternal samples. In contrast, the tuber coxae yielded a median of 0.27 × 10(6) MNC/mL BM. The first passage yielded a median of 2.19 × 10(6) MSC (1st syringe) and 1.13 × 10(6) MSC (2nd syringe) from sternal samples, compared to a significantly lower median number of MSC from tuber coxae BM (0.06 × 10(6) MSC). The number of MNC and MSC obtainable from the BM aspirates taken from the tuber coxae is significantly lower than that obtained from the sternal BM aspirates. Autologous BM for the equine athlete is particularly clinically relevant at an advanced age. Based on our findings, the tuber coxae cannot be recommended for BM aspiration in middle-aged horses.     

Upfront denileukin diftitox as in vivo regulatory T-cell depletion in order to enhance vaccination effects in a canine allogeneic hematopoietic stem cell transplantation model

Denileukin Diftitox (ONTAK^®, DAB₃₈₉ IL-2) is a recombinant DNA-derived fusion protein depleting cells that express high-affinity IL-2 receptor. Important cell targets are CD4⁺CD25⁺Foxp3⁺ regulatory T cells (T_reg). Elimination of immunosuppressive T_reg by Denileukin Diftitox may provide a way to modulate immune tolerance following stem cell transplantation. Here, we combined T_reg depletion with a vaccination approach to induce donor-specific immune reactions. To investigate this approach we chose the mixed chimerism canine stem cell transplantation model which represents a high state of tolerance between two hematopoietic systems. The aim was therefore to induce a graft versus hematopoiesis effect thereby converting mixed to full donor chimerism. Dog leukocyte antigen identical siblings that had developed a stable mixed chimerism after non-myeloablative stem cell transplantation received a single dose of Denileukin Diftitox (18 μg/kg, i.v.) followed by several cell-lysate vaccinations. Host peripheral blood mononuclear cell lysates combined with CpG-ODN, and Montanide^® ISA 51 were locally applied. In vitro studies demonstrated that canine T_reg are a target of Denileukin Diftitox. The suppression of T-cell proliferation by T_reg was abolished by addition of Denileukin Diftitox (10 nM). An increase of proliferation of median 300% (range: 200%–425%) was observed. No change in donor chimerism was observed after administration of Denileukin Diftitox and vaccination. This study highlights that application of Denileukin Diftitox resulted in a depletion of T_reg followed by an increase of immune response in vitro. This effect could not be confirmed in vivo even if the immune system was stimulated by vaccinations.