ISSN 1311-1477 (print); ISSN 131-3543 (online)
VOLUME 17, NUMBER 2, JUNE 2014
CONTENTS
Published by the Faculty of Veterinary Medicine, Trakia
University,
6000 Stara Zagora, Bulgaria
Volume 17, Number 2, June 2014, Summaries
Stefanov, I. S., A. P. Vodenicharov, M. V. Gulubova & J. R. Ananiev, 2014. Distribution of nicotinamide adenine dinucleotide phosphate diaphorase positive mast cells in the normal porcine gall bladder. Bulg. J. Vet. Med., 17, No. 2, 79–87.
The aim of the current study was to determine the localisation and number of NADPH-d mast cells in the normal domestic pig gall bladder. The gall bladders of 6 male and 6 female pigs were examined by using light microscopic enzymohistochemical and immunohistochemical techniques. Enzymohistochemistry was performed to display NADPH diaphorase activity. Then, the avidin-biotin-peroxidase complex technique for detection of mast cell tryptase expression was performed in order to confirm that NADPH-d positive cells, observed in the studied organ, are actually mast cells. Light microscopy showed strong to medium NADPH-d reactivity in the mast cells granules. Mast cells number was the highest in the propria of gall bladder fundus, body and neck as well as in the same layer of the cystic duct Ductus cysticus, followed by fibromuscular layer. The smallest density of mast cells was observed in the subserosal layer. However, statistically significant difference between the number of these cells in the muscular layer and the tela subserosa of gall bladder fundus was not detected. The density and localisation of observed tryptase positive mast cells were the same. In conclusion, the results of this study gave us a reason to presume that NADPH-d positive cells observed in the wall of porcine gall bladder and its excretory duct possess metabolic pathway for nitric oxide synthesis. Based on obtained data it is suggested that NADPH-d positive mast cells are most probably involved in the regulation of the function of epithelium, smooth muscle layer and blood vessels of the organ.
Key words: gall bladder, mast cells, NADPH-diaphorase, pig, tryptaseHematian, H., 2014. Gender-dependent micromorphometric alterations in the brain of diabetic rats. Bulg. J. Vet. Med., 17, No 2, 88-95.
This study was conducted to evaluate the effects of diabetes on the brain structure in rats and to compare the brain alterations in the two genders. Twenty adult female and the same number of male Sprague Dawley rats were divided in four groups (5 males and 5 females in each). Diabetes was induced in 3 groups by alloxan (single intraperitoneal injection of 145 mg/kg alloxan tetrahydrate). Rats with blood glucose above 200 mg/dL (11.1 mmol/L) for at least one week were considered as diabetic and were chosen for experiment. All animals were housed under the same conditions but for different periods of time (30, 60 and 90 days). Body weights were measured before and after the experiment. After the experiment, animals were anaesthesised, sacrificed and brains were collected to determine various histological parameters using histological techniques. Results revealed a decrease in male and female diabetics’ final body weight compared to that of controls. The thickness and the number of cells in the gray matter and the white matter were reduced in all test groups. In females, the thickness of white matter and the cell numbers of the gray and white matters were decreased more than in male rats. According to our results, diabetes exhibited deleterious effects on male and female rat brains, although these effects were more serious in female rats.
Key words: alloxan, brain, diabetes, gender, ratGundasheva, D., M. Andonova, P. Dzhelebov & E. Slavov, 2014. Influence of exhaustive exercise on osmotic resistance of erythrocytes and some clinical parameters in dogs. Bulg. J. Vet. Med., 17, No 2, 96-103.
The aim of the experiment was to study the in vivo effect of exhaustive aerobic exercise on osmotic resistance of erythrocytes, neutrophil/lymphocyte ratio and some clinical parameters in dogs. We used 12 male, mongrel dogs divided into two groups – animals from experimental group were submitted to exhaustive exercise; animals from control group did no exercise. Minimum osmotic resistance, 5 % haemolysis, 50 % haemolysis, 90 % haemolysis and maximum osmotic resistance were measured using an osmotic fragility test in the following dynamics: before exercise (BE), right after exercise (0 h), on 2nd hour, 4th hour, 24th hour, 48th hour, 72nd hour and on 7th and 14th day after exercise. Neutrophil to lymphocyte ratio (N/L), body temperature (BT), heart rate (HR) and breathing rate (BR) were measured in the same dynamics. We found a decrease of red blood cell osmotic resistance in experimental dogs (measured by 50 % haemolysis) on 24th hour, 48th hour and 72nd hour and on 7th and 14th day after exercise, as compared to control group (P<0.05). Surprisingly in the experimental group maximum osmotic resistance (100 % haemolysis) increased on 24th hour after exercise, compared both to initial level and control group (P<0.05). N/L ratio increased significantly in the experimental group on 2nd hour compared to BE level (P<0.05). In conclusion, exhaustive exercise acts as a stressor and affects adversely the fragility of red blood cells.
Key words: dog, exhaustive exercise, osmotic haemolysisCilev, G., z. Gacovski, a. Petreski & i. Zdraveski, 2014. Effect of partial maize substitution in fattening pig rations with by-products from vegetable and fruit processing on some carcass traits. Bulg. J. Vet. Med., 17, No 2, 104-110.
The aim of this experiment was to examine the possibilities of maize’s partial substitution as an energetic nutrient with by-products obtained by manufacturing tomatoes, peppers and grapes in the nutrition of pigs on meat yield, meatiness and the share of certain tissues in part of the carcass. The experiment was carried out on growing and fattening pigs about 60 days old and with average weight of 27.00±0.64 to 27.69±0.71 kg. Forty-eight growing and fattening pigs were divided into 3 groups, each group consisting of 8 pigs of each sex. The experiment lasted 100 days in 2 phases, 50 days each. The experimental pigs from the control group (C) were fed rations not containing any of the above mentioned by-products, whereas the two experimental groups (O-I and O-II) were fed rations where maize was substituted with 6% (O-I) and 9% (O-II) from the examined by-products. Average body weight (ABW), mass of warm carcasses (MWC) and dressing percentage (DP) of fattening pigs were 96.20 kg, 78.93 kg, 80.38% in group C, 98.10 kg, 81.68 kg, 81.60% in group O-I and 99.50 kg, 81.59 kg, 82.00% in group O-II, respectively. No significant differences were observed between the three groups (p>0.05). The values for meatiness of warm carcasses (MWC), both absolute and relative were 31.80 kg and 40.17% in group C, 33.15 kg and 40.93% in group O-I and 33.99 kg and 41.96% in group O-II, respectively. Significant difference was observed between groups C and O-II (p<0.05) for both levels (absolute and relative). The average relative proportions of bone, muscle and fat tissue in the carcass of fattening pigs were 7.45%; 42.00%; 21.62% (group C), 7.92%; 44.78%; 18.51% (group O-I) and 7.55%; 46.20%; 16.81% (group O-II). Significant difference was observed between groups C, O-I and O-II (p<0.05) were established for muscle and fat tissue contents.
Key words: meatiness, meat yield, nutrition, pigs, relative proportion of bone, muscle, fatSlavcheva, I., G. Slavchev & G. Goujgoulova, 2014. Comparative study of the rRT-PCR methods for rapid diagnosis of influenza A disease in pigs. Bulg. J. Vet. Med. 17, No 2, 111-120.
Swine influenza disease is not included in the National Prophylaxis Programme and the reports about spreading of the virus among pigs are not sufficient. The infection in pigs is usually asymptomatic, but the possible expression of clinical symptoms could result in lower productivity and economic losses. In earlier investigations concerning the spreading of swine influenza viruses (SIV) in Bulgaria, a viral circulation among the swine population was proved. For this reason and because of the high probability of emergence of new strains of unknown infectious potential, it is important to keep current diagnostic readiness in the National Reference Laboratory of Avian Influenza and Newcastle Disease Virus. The goal of the present study was to compare different protocols and combinations of kits and primer/probe sets in order to establish а rRT-PCR protocol for fast SIV diagnosis. To achieve this goal several commercial kits for rRT-PCR detection of SIV were used. After evaluation of the indicators we found that the confidence interval for the Combo M gene protocol in combination with the three used extraction kits ranged between 22.65% and 77.4%, contrasting to the combinations of used in-house amplification protocols with QIAamp Viral RNA Mini Kit (QIAGEN, Germany) and NucleoSpin RNA Virus (Macherey-Nage, Germany) extraction kits – between 67.05% and 100%. By rRT-PCR, we were able to detect RNA from all swine viruses used in this study. Our results confirmed that the correct detection of swine influenza virus is possible using a protocol different from that of the EU Reference laboratory.
Key words: Bulgaria, diagnosis, H1N1, influenza, PCR, swineAl Balaa, B., d. Zaiter, e. Hamad & a. Al mariri, 2014. Overexpression of a putative brucella melitensis zinc protease in escherichia coli induces growth arrest and cell filamentation. Bulg. J. Vet. Med., 17, No 2, 121-133.
While putative zinc protease genes have been identified in annotated sequenced genomes for many bacteria, no biochemical data have been presented to confirm that the gene product possesses protease activity. In this study, the gene encoding the putative novel zinc protease (GeneBank ID: AE008917.1) from Brucella melitensis 16M was cloned by direct PCR amplification from genomic DNA. Analysis of the deduced amino acid sequence encoded by an ORF of 927 bps revealed that this protease is a zinc metalloprotease belonging to Gluzincins family. Expression was achieved in Escherichia coli and the results demonstrated growth arrest and filamentation of E. coli cells as well as induced bacterial cell death. Proteolytic activity on skim milk and protease inhibitors effect assays confirmed that this putative protein is a member of the metalloprotease.
Key
words:
B. melitensis, cell death, cloning, protein expression, proteolytic
activity, zinc protease
Indjova, J., D. Sivrev, Kh. Fakih, M. Paskalev & Ts. Chaprazov, 2014. Repair of artificial bone defects using guided bone regeneration with Bio Oss and enamel matrix derivative. I. Light microscopic study on long bones in rabbits. Bulg. J. Vet. Med., 17, No. 2, 134–146.
Guided bone regeneration (GBR) is most effective when the material used has osteogenic, osteoinductive and osteoconductive properties. Xenografts have limited biological properties, so a solution for diversification the properties of Bio Oss® was sought by combining with Emdogain®. The purpose of the present work was to study the process of GBR involving a combination of Bio Oss and Emdogain® compared with their independent use. An in vivo experiment was conducted with 10 New Zealand white rabbits. Artificial bone defects in the tibias and femurs of each rabbit were created. Depending of the used grafting materials, six experimental groups were formed: groups 1 and 2 (Bio Oss®); groups 3 and 4 (Emdogain®) and groups 5 and 6 (Bio Oss® + Emdogain®). In control groups (A and B), osseous coagulum was used as grafting material. Prior to apposition and soft tissue suturing, all bone defects were covered with Bio Gide® membrane. Half of the rabbits (groups 1, 3, 5 and control group A) were euthanised at the end of the third, and groups 2, 4, 6 and control B – of the fourth month. There were no significant quality differences between experimental and control groups by the end of month 3 or 4. Osteoblasts, osteoclasts, osteoid and woven bone were found in all the samples studied. Maturing woven and lamellar bone were found in samples treated with Bio Oss® + Emdogain®. Newly formed bone was in close contact with Bio Oss® particles. Connective tissue around the Bio Oss® particles was found only in the groups in which it was the sole material used. There was neither evidence for degradation of Bio Oss® particles, nor for inflammation. The combination of osteoconductive properties of Bio Oss® with osteoinductive potential of Emdogain® benefited the formation of new woven bone, its transformation into lamellar and the osteointegration of Bio Oss® particles.
Key words: bone, Bio Oss®, Emdogain®, histology, osteoconduction, osteoinduction, xenograftsMahmoudi, R., p. Gajarbeygi, r. Norian & k. Farhoodi, 2014. Chloramphenicol, sulfonamide and tetracycline residues in cultured rainbow trout meat (oncorhynchus mykiss). Bulg. J. Vet. Med., 17, No 2, 147-152.
There are many concerns about safety of food contaminated with antibacterial residues. This study was designed to investigate the occurrence of tetracyclin (TC), chloramphenicol (CAP) and sulfonamide residue in cultured rainbow trout meat. A total of 100 samples of Oncorhynchus mykiss were collected from various markets in northwest regions of Iran. The prepared samples were analysed for these antibiotic residues using ELISA method. Results showed that up to 56% of the samples were contaminated with three antibiotics. The antibiotic residues concentrations in the positive samples ranged within 0.09–22.12 ng/g and the TC contamination (30%) was the highest percentage of antibiotic residues in fish meat samples. Amount of mean concentration of TC residue (8.44±6.03 ng/g) in positive samples was higher in comparison with other antibiotics. In all positive samples the TC and sulfonamide levels were below the maximum residue limit (200 and 100 μg/kg respectively). Of the 100 samples analysed for residues, CAP was detectable in only seven samples. These data showed that despite the prohibition of CAP application in food animals including aquaculture, CAP residues were detectable indicating an illegal use of this antibiotic. So, the obtained results from analysis of fish meat samples were considered to be a positive sign in terms of food safety. Also, these analyses are performed as routine according to the national residue monitoring plan of the republic of Iran.
Key words:
antibiotics residue, food safety,
Oncorhynchus mykiss
Alcigir, M. E. & S. A. Vural, 2014. Tubular intestinal adenocarcinoma in a cat: pathomorphological and immunohistochemical findings. Bulg. J. Vet. Med., 17, No 2, 153-160.
Intestinal adenocarcinoma is a malignant tumour originating from the glandular intestinal epithelium. The tumour is relatively uncommon in cats. The most affected site in cats is the ileum when compared to other species. In the case, tubular type adenocarcinoma was diagnosed in the caudal part of duodenum of 1.5-year-old female Siamese cat. Two masses of different sizes were detected on the duodenum, protruding and obstructing the lumen. Histopathologically, anaplastic glandular epithelia tried to form tubules. The anaplastic cells invaded into tunica muscularis. Mononuclear cell infiltrations were present in all layers of the duodenum. The masses were evaluated immunohistochemically by the strept ABC method and different markers (Ki67, Her2neu, CEA and p63). The first three markers were positive in anaplastic glandular epithelial nuclei, cytoplasms and membrane in the propria of the mucosa and submucosa. Ki67 and Her2neu in particular, predicted the high malignancy of anaplastic cells and poor prognosis of tumour.
Key words: feline tubular adenocarcinoma, gut, markers, pathomorphology