First record of autochthonous canine ehrlichiosis caused by Ehrlichia canis in Romania

This case study describes the first genetically confirmed and clinically manifested autochthonous Ehrlichia canis infection in a 9‐year‐old female mixed‐breed dog from Romania. Health screening of the dog included clinical examination, evaluation of stained peripheral blood smear and hematologic variables, as well as serologic testing and molecular analysis. Clinical signs included fever, apathy, dehydration, pale mucous membranes, and weakness. The microscopic examination of the blood smear and immunologic assays for Borrelia burgdorferi, Anaplasma phagocytophilum, and E canis antibodies, and for Dirofilaria immitis antigen yielded negative results. Hematologic abnormalities included moderate nonregenerative anemia, leucopenia with neutropenia, and moderate thrombocytopenia. The biochemical abnormalities identified were hypoalbuminemia, and mildly increased serum enzyme activities of AST and ALT. In addition, increased urea and creatinine levels associated with low urine specific gravity and proteinuria were also present. Nested PCR amplification of the partial E canis 16S rRNA gene demonstrated the presence of this rickettsial pathogen in the dog's blood, which subsequently was confirmed through sequencing based on the 100% homology with GenBank deposited E canis isolates. After specific treatment with doxycycline (10 mg/kg, orally, SID) for one month, the proteinuria, and hematologic and serum biochemical abnormalities with the exception of mild azotemia resolved. This report supports the geographical expansion of canine ehrlichiosis caused by E canis in nonendemic regions of Europe.     

Anatomical Variations of the Blood Vascular System in Veterinary Medicine

Traditional veterinary anatomical models describe the branches of the caudal gluteal artery as the iliolumbar, cranial gluteal, lateral caudal, satellite of the ischiatic nerve and dorsal perineal arteries. However, some classical veterinary anatomy textbooks often indicate variations the general organization of the arterial tree, without giving any pattern of origin or illustrations of the different branching. The aim of this study was to investigate the presumptive variability of the caudal gluteal artery. Two hundred and thirty‐two pelvic halves from 116 adult dogs were examined. Twelve anatomical variations were found, nine occurring in more than 5% of the dogs, and three in <5%. A ‘long‐type’ internal iliac artery, which means short caudal gluteal and internal pudendal arteries, was identified, while a ‘perineal trunk’ was observed as an interesting arterial variation. If the caudal segment alone is taken into consideration, identical vascular patterns in both hemi‐pelvises are found in 17% of the dogs. Significant statistical correlation was found for four different types of anatomic variations and gender, two types of variations and body size, one type of variation for body side and one type of variation for head shape.     

Gill Reaction to Pollutants from the Tamiš River in Three Freshwater Fish Species,Esox lucius L. 1758, Sander lucioperca (L. 1758) and Silurus glanis L. 1758: A Comparative Study

The study evaluated the effects of waterborne pollutants from the Tami? River on gill histology and possible differences in gill reaction patterns between three freshwater fish species, pike Esox lucius L. 1758, pike‐perch Sander lucioperca (L. 1758) and wels catfish Silurus glanis L. 1758 from the Tami? River. Gills from analysed fish species showed moderate to intense histopathological alterations. The most frequent progressive alteration was hyperplasia of epithelium, whereas the most frequent regressive alteration was epithelial lifting. Circulatory disturbances were most often manifested in the form of hyperaemia. During comparative analysis, differences in gill indices, reaction and alteration indices, as well as in gill and filament prevalence between analysed species, were observed. Although all analysed fish species did show both progressive and regressive alterations, there was a significant difference in the level of expression of these reaction patterns. Gill index obtained for pike clearly stands out as the lowest. Wels catfish showed the highest progressive reaction index, significantly higher in comparison with the other two species (P < 0.05), while pike‐perch showed the highest regressive reaction index, also significantly higher in comparison with the other species (P < 0.001). These results may implicate species‐specific gill reactions and thus present a useful tool for better understanding toxic mechanisms of various pollutants.     

A pre-synchronization program at early postpartum might increase the chances of Bos indicus cows cycling prior to 50 days regardless of the length of calf separation

The aim of this study was to establish if pre-synchronization would enhance the number of animals cycling prior to conventional breeding at 45 days irrespective of the length of calf separation. Multiparous Bos indicus cows were allotted in four groups (n = 10). Control group (C) dams remained with their calves; groups G24, G48 and G72, which were partially weaned for 24, 48 and 72 h, respectively, were estrus synchronized using a controlled internal drug. These procedures were performed at 25 days and again at 45 days postpartum. The number of follicles, presence of a corpus luteum and back fat thickness (BFT) were determined by ultrasound. The proportion of cows with estrus and ovulation at day 25 postpartum was statistically different between the control and treated groups, with the values being 20, 60, 50 and 70 for the control, G24, G48 and G72 groups respectively (P < 0.05). At days 45 postpartum, the proportion of cows with estrus and ovulation was different in group G48 compared with the other groups (P <0.05). The average BFT and body condition score for the four experimental groups in the two periods were similar (P >0.05). Animals with a higher proportion of follicles from 17 to 21 mm, BFT values above 3.5 mm and a regular body condition were significantly different regardless of whether the dams remained with their calves or were separated, regardless of the length of this event. It can be concluded that (1) a pre-synchronization program at day 25 could trigger the onset of ovarian activity and facilitate a breeding program at day 50 and (2) temporary weaning enhances the effect of a pre-synchronization program.     

Effect of dietary Schizochytrium microalga oil and fish oil on plasma cholesterol level in rats

The purpose of the study was to test the hypothesis that the dietary oils with different content of n‐3 polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) affect plasma lipid level in rats in a different degree. The diets with 6% of fish oil (FO) and Schizochytrium microalga oil (SchO; EPA+DHA content in the diets 9.5 + 12.3 and 2.6 + 29.5% of the sum of total fatty acids, respectively) were used; the diet with 6% of safflower oil (high content of n‐6 PUFA linoleic acid, 65.5%; EPA+DHA content 0.7 + 0.9%) was used as a control. The difference between FO and SchO was established only in the case of plasma triacylglycerol (TAG) level: plasma TAG of the FO‐fed rats did not differ from the control rats (p > 0.05), while SchO decreased (p < 0.05) plasma TAG to 46% of the control. On the other hand, FO and SchO decreased (p < 0.05) total plasma cholesterol (TC) in rats in the same extent, to 73% of the control. Regarding the underlying mechanisms for the TC decrease, both SchO and FO up‐regulated hepatic Insig‐1 gene (181 and 133% of the control; p < 0.05), which tended (p = 0.15 and p = 0.19 respectively) to decrease the amount of hepatic nSREBP‐2 protein (61 and 66% of the control). However, neither SchO nor FO influenced hepatic 3‐hydroxy‐3‐methyl‐glutaryl‐CoA reductase gene expression (p > 0.05); SchO (but not FO) increased (p < 0.05) low‐density lipoprotein receptor mRNA in the liver. It was concluded that the decrease of total plasma cholesterol might be caused by an increased cholesterol uptake from plasma into the cells (in the case of SchO), but also by other (in the present study not tested) mechanisms.     

Localization of CD9 Molecule on Bull Spermatozoa: Its Involvement in the Sperm–Egg Interaction

Tetraspanin CD9 is one of the egg membrane proteins known to be essential in fertilization process. The presence and localization of CD9 molecule in spermatozoa and its possible function in reproduction are still unclear. In our study, we describe the localization of CD9 on bull spermatozoa. In the immunofluorescence assay, the positive signal has been observed in the high proportion of sperm cells as a fine grains either on the apical part or through the entire anterior region of sperm head. CD9 recognized by monoclonal antibody IVA‐50 was detected on freshly ejaculated (83.4 ± 3.7%) and frozen‐thawed (84.3 ± 2.3%) sperm. The same reaction pattern was observed on sperm capacitated for 1 h, 2 h, 3 h and 4 h (83.6 ± 2.0%; 84.0 ± 1.5%; 85.7 ± 0.8%; 77.5 ± 10.8%). The presence of CD9 exclusively on plasma membrane of the bovine sperm has been detected by Western blot analysis of the protein fractions after the discontinuous sucrose gradient fractionation of the bull sperm. Moreover, probable role of the sperm CD9 molecule in fertilization process of cattle has been suggested as sperm treatment with anti‐CD9 antibody significantly reduced (by 25%, p ≤ 0.001) the number of fertilized oocytes compared to control group in fertilization assay in vitro.     

Expression of Mitochondria‐Associated Genes (PPARGC1A,NRF‐1, BCL‐2 and BAX) in Follicular Development and Atresia of Goat Ovaries

Most follicles undergo atresia during the developmental process. Follicular atresia is predominantly regulated by apoptosis of granulosa cells, but the mechanism underlying apoptosis via the mitochondria‐dependent apoptotic pathway is unclear. We aimed to investigate whether the mitochondria‐associated genes peroxisome proliferator‐activated receptor‐gamma, coactivator1‐alpha (PPARGC1A), nuclear respiratory factor‐1 (NRF‐1), B‐cell CLL/lymphoma 2 (BCL‐2) and BCL2‐associated X protein (BAX) played a role in follicular atresia through this pathway. The four mitochondria‐associated proteins (PGC‐1α, which are encoded by the PPARGC1A gene, NRF‐1, BCL‐2 and BAX) mainly expressed in granulosa cells. The mRNA and protein levels of PPARGC1A/PGC‐1α and NRF‐1 in granulosa cells increased with the follicular development. These results showed that these genes may play a role in the regulation of the follicular development. In addition, compared with healthy follicles, the granulosa cell in atretic follicles had a reduced expression of NRF‐1, increased BAX expression and increased ratio of BAX to BCL‐2 expression. These results suggested that changes of the mitochondria‐associated gene expression patterns in granulosa cells may lead to follicular atresia during goat follicle development.