The first record of mcr-1 gene for colistin resistance in pigs from Serbia: should we be worried?

Colistin is being used as a last-resort drug to treat infections caused by multidrug-resistant (MDR) bacteria in humans. In veterinary medicine, colistin has been used for the treatment and prevention of infectious diseases. In the first study of mcr genes by multiplex PCR in healthy pigs from Serbia, we discovered mcr-1 in 4.85% out of 350 fecal samples. The presence of mcr-1 gene was detected on three farms located less than 100 km apart from each other, predominantly in piglet samples. The results point to the necessity of monitoring of colistin resistance and the mcr genes in food producing animals as well as restricting colistin usage on farms.

Molecular diversity and evolutionary history of rabies virus strains circulating in the Balkans.

Molecular studies of European classical rabies viruses (RABV) have revealed a number of geographically clustered lineages. To study the diversity of Balkan RABV, partial nucleoprotein (N) gene sequences were analysed from a unique panel of isolates (n = 210), collected from various hosts between 1972 and 2006. All of the Balkan isolates grouped within the European/Middle East Lineage, with the majority most closely related to East European strains. A number of RABV from Bosnia & Herzegovina and Montenegro, collected between 1986 and 2006, grouped with the West European strains, believed to be responsible for the rabies epizootic that spread throughout Europe in the latter half of the 20th Century. In contrast, no Serbian RABV belonged to this sublineage. However, a distinct group of Serbian fox RABV provided further evidence for the southwards wildlife-mediated movement of rabies from Hungary, Romania and Serbia into Bulgaria. To determine the optimal region for evolutionary analysis, partial, full and concatenated N-gene and glycoprotein (G) gene sequences were compared. Whilst both the divergence times and evolutionary rates were similar irrespective of genomic region, the 95 % highest probability density (HPD) limits were significantly reduced for full N-gene and concatenated NG-gene sequences compared with partial gene sequences. Bayesian coalescent analysis estimated the date of the most common recent ancestor of the Balkan RABV to be 1885 (95 % HPD, 1852-1913), and skyline plots suggested an expansion of the local viral population in 1980-1990, which coincides with the observed emergence of fox rabies in the region.

BHK-21 cell culture rabies vaccine: immunogenicity of a candidate vaccine for humans.

Veterinary rabies vaccines produced in BHK-21/C13 permanent cell cultures have been used for a long period of time and have been proven as efficacious and safe. A candidate vaccine for human use (YU BHK Rabivak) was developed at the Pasteur Institute, Novi Sad, Serbia on the basis of the fixed rabies virus strain "L. Pasteur 2061/Vero 15 pas" using BHK 21/C13 as a cell substrate for vaccine production. To test the vaccine immunogenicity, a clinical trial was conducted involving 164 subjects between 18 and 60 years of age, immunized either with the YU BHK Rabivak vaccine candidate orwith a commercially available vaccine (Rabipur). Three groups of subjects were immunized with either vaccine by intramuscular administration in the deltoid region, following a pre-exposure regimen on days 0, 7 and 21, or the Essen or Zagreb post-exposure regimens. Rabies virus neutralizing antibodies (VNA) titres were determined by rapid fluorescent focus inhibition test (RFFIT) 21 and 30-45 days post vaccination. A protective titre of VNAs (>0.5 IU/ml) was found in all subjects vaccinated. Dynamics of the immune response showed that 96.4% of the subjects developed protective VNA titres after two doses, 99.3% after three doses and 100% after four and five doses of the candidate YU BHK Rabivak vaccine. There was a low reactogenicity without serious adverse events indicating a satisfactory safety profile in humans. Results obtained in this study indicate that BHK 21 cells offer the possibility of producing an efficacious and safe cell-culture rabies vaccine for humane use.

Osteoblastoma of the spine.

A retrograde analysis of 64 spinal osteoblastomas treated by surgery during the period 1963 - 2005 has been made. Spinal osteoblastomas were presented in 36% of all skeletal localizations. There were 44 male and 20 female patients. The average age of patients at the time of surgery was 14 years. Localizations of the osteoblastomas were cervical spine in 8 cases, dorsal spine in 16 cases, lumbar spine in 38 cases and sacrum in 2 cases. Osteoblastomas predominantly involved posterolateral vertebral elements, although in two cases a primary localization was in vertebral bodies. Neurological deficits were relatively moderate, but a paraplegia occurred in six patients. Scoliosis was present in 50% of cases, and their respective prognosis depended on the duration of the painful syndrome as well as on the age when the disease had set up. According to Enneking classification 30 patients were evaluated as stage 2 and 34 othe-rs as stage 3. Relapses were noticed in 8 (12.5%) pati-ents. Clearly delineated lesions were treated by an intralesional procedure, but aggressive ones were treated by a marginal surgery followed by additional radiotherapy (6 patients). The average follow-up period was 13 years.

[Treatment of occipitocervical instability by SUMMIT].

The occipitocervical junction with its complex anatomy and biomechanics represents unique anatomical structure difficult for operative treatment. Biomechanics forces in this region need to be resisted with rigid metallic construction to facilitate bone fusion. Surgeon must be careful about close relations of the bone, vascular and neurologic structures and must have good skills and knowledge to avoid serious complications during operations. Before 2004 for instability treatment in this region of spine wires and pin construction for fixation of bone grafts were used. In our Institute rod constructions with screws have been used to get more rigid construction since 2004. In this paper we will present our early results in occipitocervical instability treatment by SUMMIT instrumentation.

[The treatment of infected diaphyseal femoral defects by lengthening one of the bone fragments by Ilizarov].

We analyzed 30 patients with infected diaphyseal defect of femur, which have been treated by lengthening one of the bone fragments with Ilizarov apparatus. The mean length of the bone defect was 6 cm. Substitution of the defect, bone healing and elimination of the infection was achieved in 27 patients. The mean time of apparatus fixation was 10 months. According to Palley scoring system, 10 patients had excellent functional results.

[Differences in the rehabilitation period following two methods of anterior cruciate ligament replacement: semitendinosus/gracilis tendon vs ligamentum patellae].

In this study we have analyzed outcome during the early rehabilitation period phase following two different methods of anterior cruciate ligament (ACL) reconstruction: ligamentum patellae (LP) and semitendonosus/gracilis tendon (SG) based reconstruction. This study included 40 patients treated by each method, examined 6 weeks and 3, 6 and 12 months after surgery. Patients in the SG group showed significantly better Lysholm scores at 6 and 12 months, Tegner Activity Scale scores at 3 months, and pain profile assessments at 6 weeks and 3 months than those in the LP group. Significant differences were observed in LP group in range-of-motion at 6 weeks and 3 and 6 months post-surgery. Stability tests revealed no significant differences between patients in the two groups. SG-based reconstruction of the ACL thus demonstrated advantages over LP-based reconstruction regarding pain and function, while LP-based reconstruction was associated with an earlier return of motion.

Low back pain--differential diagnosis, prognosis and treatment.

From January 2002 to February 2003, 137 patients complaining of low back pain were treated at the Institute for Orthopedic Surgery "Banjica", Belgrade, Serbia. There were 89 male and 48 female patients aged 13 to 77, mean age 42.2. Their condition was diagnosed through use of radiography, CT, MRI, EMNG, standard battery of neurological tests, and laboratory analyses (urine and blood analysis). Surgical treatment was performed on 39 patients; all other patients received some form of non-surgical care (physical therapy, medication or corset). Treatment efficacy was evaluated by use of the visual analog scales (VAS) and the Oswestry index, before and after treatment. The use Wilcoxon's pair test revealed statistically significant difference between before and after treatment data on VAS and Oswestry index for all patients.

[Treatment of bite wounds and cat-scratch disease]. / Tretman ujednih rana i bolest macjeg ogreba.

Although rabies is the most serious consequence of animal bite injuries, in urban rabies-free countries the risk of rabies is far lower than the risk of bacterial infections of the wound. The most frequent etiologic cause of the wound infections after dog bites is Pasteurella multocida, as well as in the case of bites by cats, after which cat-scratch disease may also develop, its main cause being Bartonella henselae. All bite injuries must be carefully cleaned and disinfected; it is necessary to estimate the need of antirabies and antitetanus protection, and to introduce antibiotic treatment for prevention, particularly in the case of deep sting wounds caused mostly by cats.

[Evaluation of the Respiralex test for rapid isolation of Streptococcus pyogenes directly from throat swabs]. / Evaluacija Respiralex testa za brzo dokazivanje Streptococcus pyogenesa direktno u brisu guse.

Results obtained by fast Respiraplex test were compared with findings of classical method of cultivation in order to assess possibilities of this test in isolation of Streptococcus pyogenes directly in the throat swab. The sensitivity of the fast test was 79.4%; specificity 100%. On the basis of gathered results, the following can be concluded: the fast test cannot completely substitute the classic method of cultivation although high specificity occurs, because of low sensitivity especially in patients with small number of colonies in the culture.

[Prevention of human rabies in Yugoslavia]. / Profilaksa besnila ljudi u FR Jugoslaviji.

Over the last two decades human rabies prophylaxis in Yugoslavia was successful and not a single case of the disease was registered. However, every year there are registered cases of those injured by animals in which rabies was confirmed by laboratory evidence. In FR Yugoslavia every year 800-1000 people are vaccinated against rabies, which makes 10% of all the people injured by animals in Yugoslavia. During 1997 there were 10.266 patients injured by animals; 816 (7.95%) were vaccinated whereas 52 were injured by rabid animals or rabies-suspected animals; 662 were injured by unfamiliar or wild animals, while 102 patients were injured by familiar or preventively vaccinated animals. In other patients vaccination was avoided by a 10-day veterinarian surveillance of the dog. New literature estimated application of reduced so-called "Zagreb-2-1-1" vaccination protocol in severe injuries as unsuccessful in children because of active immune response suppression by simultaneously given immunoglobulin, and that is why this protocol must be abandoned.

[Knowledge of rabies in schoolchildren]. / Znanje skolske dece o besnilu.

Rabies is a zoonosis which rarely occurs in our country, mostly due to good control of this disease. In order to assess knowledge of final grade elementary schoolchildren about rabies, a poll was conducted in Novi Sad (town) and Curug (village) in 1997. Gained results show that a great number of children have household pets, much more in rural than urban areas. Although most children have knowledge about rabies prevention, knowledge of a certain number of children is not sufficient and they do not have proper hygiene habits in contact with animals. Taking into consideration the fact that dog population increases both in urban and rural areas, corresponding education of their owners, including children, is necessary. Health education is the method of choice in health protection and promotion of health, skills and behavior in regard to health and possible health risks engaged in contacts with domestic animals-household pets, whereas, apart from the family, the school must play an important role.

[Non-specific sacroiliitis]. / Nespecificni sakroileitis.

We present retrospectively five cases of non-specific sacroileitis which were established and treated in Special Orthopaedic Hospital Banjica Belgrade in the period of 12 years, from 1982 to 1994. Patients have been from 17 to 27 years of age. All of them had been treated initially in other institutions under different diagnosis: specific sacroileitis, specific coxitis, specific spondylitis, abscess of gluteal region and discarthrosis and lumboishialgia. All of the patients have been treated by antibiotics, one by tuberculostatics, one surgically by drainage and one by coxofemoral spica cast. In spite of short period of follow up between 1.5 month and 2 years, we have had excellent clinical and laboratory recovery.

[Clinical trial of YU BHK Rabivak vaccine against rabies in volunteers]. / Klinicko ispitivanje YU BHK RABIVAK vakcine protiv besnila na dobrovoljcima.

The BHK (baby hamster kidney) continual cell line has been used for years in the production of rabies vaccine for animals. Becouse of the rich harvesting of rabies virus from this cell substrate, there is no need of the an additional virus concentration, so that the vaccine production can be organised in small local Pasteur Institutes as well. Although its long-range use in the veterinary medicine has proved that the BHK line is safe and that the vaccine is immunogenic, its use in the human medicine has been only recently studied. According to the latest recommendations of the Expert Committee for the Biological Standardisation of the WHO from 1998, residual cellular DNA in the vaccine is not a potential risk because the inactivation of the vaccine by the beta-propiolactone completely inactivates its possible biologic activity. In the work, the authors report the results of the third phase of the clinical trail of the YU BHK Rabivak vaccine on volunteers. The rabies vaccine studied, named YU BHK Rabivak, produced by Novi Sad Pasteur Institute, caused the generation of protective titre of virus-neutralising antibodies in all examinees: in 96.4% after 2 doses, in 99% after 3 doses, and in 100% after 4 and 5 doses of the vaccine. The registered side effects were expected, mild, did not need medical treatment, and were not more frequent than in the case of other vaccines from cell culture.

Immunogenicity of BHK-rabies vaccine in human volunteers.

The production of rabies vaccine on baby hamster kidney (BHK-21) cells for human use is discussed. Long term experience in application of this vaccine in animals, without any noticeable complications and findings of inactivation of contaminated DNA from the cell substrate by beta-propiolactone have justified its recommendation for human use. Preliminary results of applying this simple, adjuvant vaccine in volunteers, confirmed its good tolerability and immunogenicity.

[Rabies in dogs and efficacy of active immunoprophylaxis in Yugoslavia]. / Besnilo pasa i efikasnost aktivne imunoprofilakse u FR Yugoslavia.

In this study we analyzed 36 cases of dog rabies diagnosed at the Pasteur Institute in Novi Sad during the last 10 years as well as the quality of postvaccination immunity. Most dogs had the paralytic form, more common in sylvatic rabies than furious clinical picture. 11% of dogs were aggressive, whereas 44% had injured humans, including their owners. Postvaccination immunity was examined in a group of dogs vaccinated with three domestic and one vaccine imported from abroad. Vaccination was successful in more than 70% of dogs and it is a sufficient percentage of seroconversion in dog rabies eradication. However, in dogs vaccinated only once during lifetime, the immunity was not satisfactory. That is why in primary vaccination of dogs compulsory revaccination 30 days after the first vaccine dose is suggested.

[Neurotrichinosis]. / Neurotrihineloza.

INTRODUCTION: Trichinellosis is an anthropozoonosis caused by Trichinella spiralis. Central nervous system complications can occur during the course of this disease. This paper presents a case of neurotrichinellosis, assayed by using indirect immunofluorescence test. CASE REVIEW: A patient aged 30, was admitted to the Clinic on the fifth day after onset of the disease, which was gradual, starting with flu-like symptoms. Two days before admittance, the diseased became somnolent, disconcerted and disoriented. On admission, the patient was highly febrile (39 degrees C), dehydrated, with eyelid edema. Right hemiparesis was present. The disease was confirmed by indirect immunofluorescence test, showing an increase of trichinella antibody titre (1:20, 1:160 and 1:640). The cerebrospinal fluid was cytobiochemically normal. Electroencephalographic findings exhibited a moderate cerebral dysfunction. Multifocal unspecific changes were established by magnetic tomography. The diseased was treated by mebendazole and prednisolone. The course of the disease was favourable and the patient was cured without sequelae. DISCUSSION: Recognition and diagnosis of Trichinellosis are complicated due to its polymorphid symptomatology. In the case reviewed, the disease started with clinical features of flu-like symptoms and febrile gastroenteritis. According to some authors, central nervous system manifestations occurred in 10-15% of the diseased. CONCLUSION: When examining clinically manifested encephalitis, Trichinelosis should be taken into consideration as a cause of the disease.

[Determination of nutritional levels in patients on chronic hemodialysis programs using anthropometric methods]. / Odredivanje stepena ishranjenosti bolesnika na hronicnom programu hemodijalize antropometrijskim metodama.

The results of a number of methods used in the cross-sectional nutritional assessment differ markedly in patients on regular hemodialysis (RHD). Correlations of nutritional parameters were investigated in 31 patients on RHD and in 31 control subjects with similar sex and age structure without any considerable difference in any of the examined nutritional parameters. In the patient group on RHD, beside a high mutual correlation of general nutritional parameters and a high correlation of general parameters with fat compartment parameters and the fat free compartment (protein) parameter, there is also a high correlation of fat parameters with the protein parameter which does not occur in the control group. This can possibly explain the difficulties in the nutritional assessment of patients on RHD. Out of this reason we suggest the usage of the RBM/F classification which is a combination of relative body mass values (RBM) and the body fat content (F) percentage.

[Nutritional levels and in vitro tests of thyroid function in patients on a chronic hemodialysis program]. / Stepen ishranjenosti i in vitro testovi tireoidne funkcije kod bolesnika na hronicnom programu hemodijalize.

The nutritional status of 31 patients on regular hemodialysis (RHD) and of 32 control subjects was assessed by the Simic RTM/M classification, and in vitro thyroid function tests were determined in each subgroup. There were 13 malnourished, 13 normally nourished and 5 obese patients on RHD and 9 malnourished and 23 normally nourished control subjects. Differences between subgroups were considered in the light of their sex and age structure, duration of regular hemodialysis, proportion of patients with concomitant chronic illness, and the proportion of patients which received transfusion within the last 7 days. TT4, FT4 and rT3 were influenced by the nutritional status of the patients on RHD, while TBG, TT3 and FT3 were not. The statistical significance between RHD patient subgroups and the subgroups of control subjects was altered only in the case of TT4.

[Manifestations of visceral and ocular symptoms of toxocariasis in a 6-year-old boy]. / Pojava visceralne i okularne klinicke slike toksokarijaze kod sestogodisnjeg decaka.

In the past 10 years we have examined 137 cases of toxocariasis, predominantly in children. Three cases were with unilateral ocular involvement. The article reviews a 6-year-old boy with left side strabismus and granulomatous chorioretinitis. Laboratory examinations revealed blood eosinophylia 24% and IgG against Toxocara canis larvae in titer 1:320 by indirect immunofluorescent assay. In epidemiologic anamnesis we concluded that about 4, 5-year-long geophagia was the source of infection.