Isolation and identification of Brucella suis biotype 2 from epididymal puncture performed on a boar affected with brucellosis

The causal agent of swine brucellosis is Brucella suis. Within the scope of the kinds of Brucella suis, there are five biotypes, but only biotypes 1, 2 and 3 lead to swine infections. Human infections with Brucella suis biotype 2 are rarely registered. Swine brucellosis is widespread all over the world. It has been noted that the incidence of swine population infected with Brucella suis in Western Europe has been increasing during the recent years. The goal of this project was to isolate, identify and typify the causal agent from epididymal puncture performed on a boar with conditions suspicious of brucellosis, using standard microbiological methods. The results of the research show that Brucella suis biotype 2 can be successfully isolated and identified from a sample obtained by means of epididymal puncture of live animals. Therefore, epididymal puncture gives us a certain, reliable and important sample derived from a live animal for a direct diagnostic of boar brucellosis. The above mentioned first isolate of Brucella suis biotype 2 epididymal puncture, has been marked as K-1

Correlation between Different Helicobacter Morphotypes and Histological Changes in Pig Gastric Mucosa

Background: Two distinct morphologic types of bacteria which belong to the Helicobacter species, have been described in pigs: once or twice curved Helicobacter pylori-like bacteria (HLO) and the multicoiled, Gastrospirillum-like bacteria (GLO). The objective of this study was to determine the incidence of Helicobacter spp. using modified Giemsa stain and to define the relationship between presence of Helicobacter spp. and histopathological changes of gastric mucosa in pigs.Materials, Methods & Results: A total of 120 pig stomachs (60 from intensive and 60 from extensive breeding) were enrolled in this study and 240 fragments of fundic and pyloric mucosa were taken for histopathological examination. By modified Giemsa staining of gastric mucosa, Helicobacter-like organisms were confirmed in 4/60 (6.67%) of pigs in intensive and 5/60 (8.33%) of pigs in extensive breeding. The incidence of tightly spiral shaped Gastrospirillum-like organisms in pigs of intensive and extensive breeding were 5/60 (8.33%) and 9/60 (15%), respectively. The severity of gastritis was scored to the Sydney System with some modifications. There was no significant difference between HLO-positive and HLO-negative fundic mucosa in pigs of both breeding systems. In contrary, there was significant difference between HLO-positive and HLO-negative pyloric mucosa of pigs in intensive (P < 0.001) and in extensive breeding (P < 0.05). In intensive breeding, there was significant difference between GLO-positive and GLO-negative fundic mucosa (P < 0.05), while there was no significant difference between GLO-positive and GLO-negative pyloric mucosa.Discussion: The prevalence of Helicobacter spp. bacteria were in coherence with literature data. The higher prevalence of both morfological type of bacteria, were considered in pigs in extensive breeding. the hygienic conditions and managment factors in pigs farm are the possible impact for higher bacterial transmission. The association of high prevalence of H. pylori and poor hygienic condition was shown by epidemiological studies conducted on humans. In both, humans and pigs, the presence of H. pylori correlates with an inflammatory response, but there are differences in inflammatory cell population. In H. pylori infected humans, neutrophils composed the bulk of cellular infiltrate, while in pigs, the primary inflammatory cell was the lymphocyte, which is in accordance to results published by others autors. Thus it indicates that different hosts exhibit a different pathohistological response to the Helicobacter spp. infection. In human as well as in veterinary pathology, the fact of the different pathogenicity of various Helicobacter species is well known. In all HLO-positive pyloric mucosa, moderate to severe focal or diffuse infiltration of mononuclear cells and lymphoid follicles with germinal centers, were observed. A similar conclusion was drawn from results of an experimental infection study in pigs. There was signifficance between HLO-positive and HLO-negative pyloric mucosa in both, intensive and extensive breeding. In the contrast, GLO were not associated with the presence of severe gastritis, but only with mild to moderate superficial infiltration of lymphocytes and plasma cells in both, fundic and pyloric mucosa. There was no significant difference between GLO-positive and GLO-negative pyloric mucosa of pigs in both breeding systems. Despite the low gastritis score of fundic mucosa in pigs in intensive breeding, there was a significance difference between in GLO-positive and GLO-negative fundic mucosa. It is believed that the possible reason of such results is the meals with low fiber content and low particles size. These results suggest that the presence of HLO, but not of GLO is associated with the pyloric gastritis in pigs

Analysis of Different Diagnostis Methods of Influenza in Horses

Background: Equine Influenza is a serious, acute respiratory illness with characteristical clinical signs. The disease is caused by family of Orthomyxoviridae, genera Influenza virus A by two subtypes H7N7 and H3N8. Currently, there is believe that H7N7 has been replaced as a predominant subtype with the H3N8. Horse infection with influenza virus can be detected by serological tests on paired sera using HI test. Commercial rapid tests could be used for the detection of influenza virus. Recently it is widely use a PCR method as fast and more specific methods.Materials, Methods & Results: Fifty horses and one pony, age between one and 22 years have been included in experiment. Horses were of different race, sex, and age and vaccination status. Ten out of total 51 (10/51) have been regularly vaccinated against EI. Prior to initiation of these study epidemiological survey has been performed. The clinical examination has been followed by blood sampling for blood cell and serum extraction. The serums were evaluated by HI method. Nasal swabs are taken from both nostrils twice, one was frozen for virus detection by RT-qPCR while another was used for detection of EI virus by Directi-gen™ FLU A rapid test. Analysis of titers of antibody reveled that 7 horses (14%) had specific antibodies (IgG) against subtype H7N7, while 9 horses (18%) had specific antibodies against H3N8. In the same time 4 horses had specific antibodies against both subtypes. Serological data confirmed that from 48 horses (96%) had the titer of antibodies greater than 16 against H7N7, while 40 horses (80%) had the specific antibodies (IgG) against H3N8. We found quite unexpected presence of specific antibodies (IgG) for H7N7 in horses that have not been previously vaccinated with H7N7 subtype. These horses never been utilized for sport activity and there was no legal requirements for their vaccination. Could horses with specific antibodies for H7N7 be transfected from vaccinated horses we could not confirme with scientific evidence either we could not have evidence that wild and domestic birds played a significant role especially knowing that horses are dead end of further we could confirm it. Our findings unequivocally confirmed that EI virus subtype H7N7 antibodies (IgG) were present in horses that have not been vaccinated and this is serological evidence that virus H7N7 is circulating  in these geographical areas.Discussion: Fast and reliable diagnosis and isolation of suspicious horses represent the first line of defense against pandemic influenza. Recognition of clinical signs (fever, depression, sharp cough and nasal discharge) along with epizootical survey provides the basis for the early detection of infection. In some cases, cough and rapid spread of symptoms of cough in a group of horses that were either unvaccinated against influenza virus or have been in contact with influenza virus infected horses can clearly point out to the EI virus. The definitive confirmation of EI virus could be done by virus isolation on tissue culture or embrionated eggs followed by detection of virus nucleic acid by RT-PCR. Our data suggest that a substantial number of horses (90%) that have not been vaccinated or vaccinated irregularly had specific antibodies against both subtypes of EI, what suggest that those horses have been exposed to viruses sometimes during their lifetime. Additionally, despite the fact that 20% of horses had some signs of respiratory disease that resemble EI we were not able to confirm EI nucleic acid by RT-PCR while Directi-gene™ assay confirmed virus presence just in two horses. Failure to detect virus nucleic acid could be due to fact that nasal swab samples have been taken at the end of the clinical symptoms, other authors have simular PCR negative patient which displayed a significant rising titre to influenza type A

Entomological and ecological index for risk of infection causing lyme disease in territory of Vojvodina, Serbia

In Europe, of all the vector transmitted diseases, the occurrence of lyme disease is the one most often registered, and the most significant vector Borrelia burgdorferi is the tick Ixodes ricinus. Both humans and animals contract lyme disease. The risk of the occurrence of lyme disease is in correlation with potential exposure to tick bites and depends on the density of the tick population in the endemic area, the percentage of ticks infected with the cause of lyme disease, the duration and the nature of the activity of the susceptible population in a certain area. The objective of these investigations was to determine the entomological and the ecological risk index, as well as to assess the risk of transmission of the cause of lyme disease in the territory of Vojvodina Province in the Republic of Serbia. Ticks were collected at 12 locations in the South Bačka District of Vojvodina. A total of 1400 ticks were identified up to the level of species. After establishing the infection of ticks with the cause of lyme disease, the entomological and the ecological index was determined for the given regions using microscopic examination in a dark field. Two species of ticks aere identified in this geographic region (Ixodes ricinus and Dermacentor marginatus). Examining I. ricinus, the prevalence of infection B. burgdorferi was established, ranging up to 33.1%. The ecological risk index indicates that there is a potential risk of humans and animals becoming infected at 8 localities. It was determined for 3 localities that there is a definite actual risk of the transferrence of causes of lyme disease

Development of a prototype of an ambient-aware two-arm mobile service robot

This paper describes a prototype of an ambient-intelligent advanced service robot of anthropomorphic characteristics that is intended for operation in indoor environment as well as for safe interaction with people. The robot consists of a wheeled mobile platform with a spinal (segmented) torso, bi-manual manipulation system with hands, and a robot head with capabilities to see, hear and speak. It is equipped with a number of advanced sensors, including indoor laser range finder, several ultrasonic probes as distance sensors and obstacle detectors, 3-axis inertial sensors with gyroscope, stereo vision system, 2 wide-range microphones, and 2 speakerphones. Its operation is autonomous but it may be controlled from a host computer through a wireless link. The robot prototype is expected to express advanced cognitive capabilities including spatial understanding, autonomous motion, affective and social behavior. The development of the robot is a joint effort of four Serbian academic institutions and it is expected to have it fully operational in the second half of 2015

Relationship of Circulating Tumor Necrosis Factor Alpha (TNF-α) and Insulin Secretion and Resistance in Euglycaemic Dogs

Background: Insulin resistance is a state that is characterized with reduced sensitivity of peripheral tissues to insulin. It can be related with increased level of tumor necrosis factor alpha (TNF-α) in dogs. Insulin resistance can be evaluated by homeostasis model assessment (HOMA-IR, HOMA-β). The aim of this study was to determine correlation of circulating TNF-α level with insulin production and insulin resistance indexes in euglycaemic dogs. Materials, Methods &amp; Results: Seventydogs of normal body score were included in this study. After blood sampling levels of glucose, insulin and TNF-α were determined and indexes HOMA-IR and HOMA-β were calculated. Three groups in accordance to TNF-α levels were formed: the first-TNF-α 0-2.0 pg/mL, the second-TNF-α below median (2.1-17.0 pg/mL) and the third-TNF-α above median (17.1-51.8 pg/mL). Differences in insulin and glucose levels, HOMA-IR and HOMA-β were determined in all three groups. ANOVA and posthock LSD analyses were used. Correlation between HOMA-IR and HOMA-β was determined. Linear regression between HOMA-β/HOMA-IR ratio and glucose concentration was calculated. SPSS statistical program was used (IBM). Highest insulin level was detected in the second group and the lowest was detected in the third group. The lowest glucose level was detected in the first group. The highest value of HOMA-β index was noted in the first group and it decreases with TNF-α increase. The highest HOMA-IR value was detected in the second group and the lowest was in the third group. Positive correlation was noted between HOMA-IR and HOMA-β. Significant linear correlation was noted between glucose levels in function of HOMA-β/HOMA-IR (R2= 0.51-0.78, P = 0.0007). The first group showed the minor change of glucose level (b= 0.29 mmol/L). In the third group the greatest change of glucose level in function of HOMA-β/HOMA-IR was noted (b= 0.52 mmol/L). In the third group the highest increase of glucose level followed by decrease in HOMA-β/HOMA-IR ratio was noted.  Discussion: Increase of TNF-α followed by increase of insulin was noted in the second group. That indicates reduced insulin action and compensatory increase in his concentrations in order to achieve the same effect. TNF-α induces reduction in expression of glucose transporter 4 that is insulin-regulating hormone. Serine phosphorylation of insulin receptor substrate-1 that is induced by TNF-α causes inhibition of insulin receptors. These actions cause insulin resistance and compensatory increase in insulin secretion. Increased tissue resistance is reflected in increased HOMA-IR index which is directly associated with insulin and glucose level. Increase of insulin value and HOMA-IR index were noted in second group. This indicates the influence of TNF-α on insulin resistance. The lowest insulin level was noted in the third group of dogs. Decrease in production and secretion leads to reduction in circulating insulin and can be evaluated by HOMA-β index. The highest value of this index was noted in first group and decreases with TNF-α increase. This can be related with apoptotic effect that TNF-α has on beta- pancreatic cells. Increase of HOMA-β index causes increase of HOMA-IR. This is indication of increased peripheral resistance. Compensatory mechanism for that state is increased insulin secretion. Glucose level will increase more during increase of HOMA-IR and decrease of HOMA-β. This indicates that TNF-α regulates glucose level directly and explains differences in glucose concentrations in dogs with different concentrations of TNF-α showed. Values of insulin resistance indexes, glucose and insulin were affected by circulating concentration of TNF-α. The most unfavorable change in glucose concentration based on insulin production and tissue resistance was founded in dogs with the highest circulating TNF-α concentration in blood.

Seroprevalencija infekcije izazvane uzročnikom Anaplasma phagocytophilum kod pasa u Autonomnoj Pokrajini Vojvodini, Srbija.

Granulocytic anaplasmosis in dogs is a disease that is distributed worldwide, caused by the pathogen Anaplasma phagocytophilum. This disease is transmitted by ticks of the Ixodes genus. So far, no data have been published about the presence and prevalence of dog infections with A. phagocytophilum in the Autonomous Province of Vojvodina and the Republic of Serbia proper. The aim of this seroepidemiological research was to determine the seroprevalence of IgG antibodies to the agent A. phagocytophilum in the population of dogs in Vojvodina, Serbia. This seroepidemiological research involved 84 randomly selected dogs from the area of the Autonomous Province of Vojvodina. Aiming to determine the presence of antibodies of class G to the agent Anaplasma phagocytophilum, we used the indirect immunofluorescence antibody test (IFAT). By applying IFAT, in the area of the Autonomous Province of Vojvodina, Serbia, specifi antibodies of class G to agent A. phagocytophilum were found in the serums of 13/84 dogs, which points to a seroprevalence of 15.5 %. The detection of antibodies to the agent A. phagocytophilum in the dog population in the area of the Autonomous Province Vojvodina is the basis for further epidemiological, clinical and diagnostic research of this infection.Granulocitna anaplazmoza pasa proširena je diljem svijeta, a prouzročena je vrstom Anaplasma phagocytophilum. Prenosi se krpeljima roda Ixodes. Do sada u Autonomnoj Pokrajini Vojvodini i Republici Srbiji nema objavljenih podataka o prisutnosti i raširenosti infekcije pasa uzročnikom A. phagocytophilum. Cilj je ovoga seroepidemiološkog istraživanja da se utvrdi seroprevalencija IgG protutijela za A. phagocytophilum u populaciji pasa u Vojvodini, Srbija. Ovim seroepidemiološkim istraživanjem, obuhvaćena su 84 nasumično izabrana psa s područja Autonomne Pokrajine Vojvodine. U cilju utvrđivanja prisutnosti protutijela razreda IgG za A. phagocytophilum rabljen je test indirektne imunofl uorescencije (IFAT). Primjenom IFATa, na području Autonomne Pokrajine Vojvodine, Srbija, utvrđena su specifična protutijela IgG na uzročnika A. phagocytophilum u serumu 13 pasa, što predstavlja seroprevalenciju od 15,5 %. Dokaz protutijela za A. phagocytophilum u populaciji pasa na području Autonomne Pokrajine Vojvodine osnova je za daljnja epidemiološka, klinička i dijagnostička istraživanja ove infekcije

Slučaj humane monocitne erlihioze u Srbiji

Introduction Ehrlichiosis is a bacterial zoonosis transmitted by hematophagous arthropods - ticks. In humans, it occurs as monocytic, granulocytic, and ewingii ehrlichiosis. Pathological process is based on parasitic presence of Ehrlichia organisms within peripheral blood cells - monocytes and granulocytes. Case Outline Fifty-two year old patient was admitted to hospital due to high fever of over 40°C that lasted two days, accompanied with chills, muscle aches, malaise, loss of appetite, headache, confusion, breathing difficulties, and mild dry cough. The history suggested tick bite that occurred seven days before the onset of disease. Doxycycline was introduced and administered for 14 days, causing the disease to subside. Indirect immunofluorescence assay was used to analyze three serum samples obtained from this patient for Ehrlichia chaffeensis antibodies, and peripheral blood smear was evaluated for the presence of Ehrlichia and Ehrlichia aggregation into morulae. Conclusion Ehrlichiosis should be considered in each case where there is a history of tick bite together with the clinical picture (high fever, chills, muscle aches, headache, generalized weakness and malaise, and possible maculopapular rash). The presence of Ehrlichia chaffeensis antibodies was confirmed in a patient with the history of tick bite, appropriate clinical picture and indirect immunofluorescence assay. This confirmed the presence of human monocytotropic ehrlichiosis, a disease that is uncommonly identified in our country.Uvod Erlihioza je bakterijska zoonoza koja se prenosi hematofagnim artropodama - krpeljima. Kod ljudi se javlja kao monocitna, granulocitna i ewingii erlihioza. Patološki proces je posledica unutar ćelijskog parazitiranja erlihije u monocitima i granulocitima periferne krvi. Prikaz bolesnika Bolesnik star 52 godine primljen je na odeljenje zbog visoke dvodnevne febrilnosti (preko 40°C) koja je praćena drhtavicom, bolovima u mišićima, malaksalošću, gubitkom apetita, glavoboljom, konfuznošću, otežanim disanjem i oskudnim suvim kašljem. U anamnezi je dobijen podatak o ujedu krpelja sedam dana pre prijema. Laboratorijski nalazi su ukazali na trombocitopeniju, leukopeniju, anemiju i povećanje aktivnosti transaminaza u serumu. Bolesnik je lečen doksiciklinom 14 dana, nakon čega su se tegobe povukle. Metodom indirektne imunofluorescencije analizirana su tri uzorka seruma ovog bolesnika na prisustvo antitela na bakteriju Ehrlichia chaffeensis i pregledan je uzorak razmaza periferne krvi na prisustvo erlihija i konglomerata erlihije u morule, koje predstavljaju citoplazmatske vakuole. Zaključak Kod bolesnika s podatkom o ubodu krpelja, odgovarajućom kliničkom slikom i serološkim testom indirektne imunofluorescencije dokazana su antitela za bakteriju Ehrlichia chaffeensis, što ukazuje na humanu monocitotropnu erlihiozu, bolest koja se kod nas retko dokazuje. Na erlihiozu treba misliti kada uz kliničku sliku (visoka febrilnost, groznica, bolovi u mišićima, glavobolja, opšta slabost i malaksalost, eventualno makulopapulozna ospa) postoji podatak o ubodu krpelja