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During the years from 1993 to 2000, 183 strains of Pseudomonas aeruginosa were isolated from different pathological specimens originating from dogs. Antimicrobial susceptibility patterns against 10 antipseudomonal agents were obtained on 183 P. aeruginosa strains. In vitro antimicrobial susceptibility testing was performed using the disk diffusion method (Kirby-Bauer). Antimicrobial susceptibility profiles showed that among β-lactam antibiotics, imipenem was the most active compound. Out of the 183 strains tested, 96.7% were sensitive to imipenem. Cefoperazone showed good in vitro activity against 86.9% of the tested strains. Against ceftazidime, 77.0% of strains showed sensitivity. An old penicillin, carbenicillin, gave only 71.6% sensitive strains. Sensitivity to amikacin was 87.4% and it was 83.1% to gentamicin. Pipimedic acid, a first-generation quinolone, was the least active compound of all those tested, 47.0% were resistant. The in vitro sensitivity against enrofloxacin showed that 71.0% strains were sensitive and 26.2% showed resistance. Almost all strains tested, 93.4%, were susceptible to ciprofloxacin and marbofloxacin. Besides imipenem, the quinolone antibiotics, marbofloxacin and ciprofloxacin were the most effective against P. aeruginosa strains isolated from dogs. [ABSTRACT FROM AUTHOR]

Background/Aims. Acute appendicitis (AA) remains a diagnostic challenge in children, despite ongoing researches. With an aim to facilitate making diagnosis of AA many scoring systems have been created; among them Alvarado score is the most popular. C-reactive protein (CRP) has proven significance for diagnosing AA in adults, but not in children. The aim of this study was to evaluate significance of Alvarado score, as well as CRP values, in making diagnosis of AA in children. Methods. This prospective six-month study was performed on 257 patients under the age of 15, admitted for acute abdominal pain in the Clinic of Pediatric Surgery and Orthopedics in the Clinical Centre of Niš. Alvarado score and CRP values were determined on admission and compared with final diagnosis on discharge. The patients were divided into two groups: group I - non operated patients with abdominal pain (n = 184) and group II - operated on patients for appendectomy (n = 73). Results. Values of Alvardo score were statistically significantly different between groups (group I: 4.9 ± 1.21, group II: 8.55 ± 1.32). Also, our results showed significantly high values of CRP measured in operated children (group I: 8.17 ± 4.7 mg/L, group II: 38 ± 26 mg/L). Values of validity parameters for Alvarado score were: sensitivity 90%, specifity 80%, positive predictive values 87%; for CRP 95%, 70% and 80%, respectively. Conclusion. Alvarado score and CRP are very useful adjuvant diagnostic tool for AA to a less experienced surgeon. High values of Alvarado score and CRP cannot be ignored neither at the same time, used as the sole diagnostic method for discriminating children with AA.

Transverse testicular ectopia is an extremely rare anomaly, characterized by migration of one testis towards the opposite inguinal canal, usually associated with inguinal hernia. Spermatic cord of the ectopic testis originates from the appropriate side. In most reported cases, the accurate diagnosis has not been made before surgery. This is a case report of transverse testicular ectopia in eleven-year-old boy who had undergone an operation for the left inguinal hernia in age often months. At the time of herniorrhaphy, the right testis was absent. Ten years later, during re-operation of the left inguinal hernia, both testis were found in left inguinal canal and easily brought down sequentially through the left groin into the scrotum. The right testis was fixed in the left hemiscrotum, due to shorter funicular elements, and the left was trans-septally moved to the right hemiscrotum (a modified Ombrédanne operation). Ultrasonography and voiding cystoureterography showed no associated genitourinary anomalies and no Mülerian duct remnants. The rupture of gubernaculum and dysfunction of the genito-femoral nerve could explain the etiology of crossed testis ectopia. Although ectopic testis could be localized preoperatively by ultrasonography, CT, MRI, arteriography and venography, correct diagnosis was made intraoperatively in the majority of cases. Treatment modalities include laparoscopic and surgical procedures. Atrophie testis should be removed. If testes are fused, they have to be brought into one hemiscrotum. In cases where testes are completely separated with individual funicular elements and vas deferens, an ipsilateral or contralateral orchiopexy should be performed depending on the length of funicular elements.

Abstract Background Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described. Findings Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells. Conclusions Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.

Molecular methods are increasingly being utilized for accurate identification of ticks (Acari: Ixodidae), especially in cases of morphologically highly similar species. In this study, we performed molecular research of the tick fauna in Croatia using DNA barcoding method. Ticks were sampled in three biogeographical regions and thirteen species were recorded: Dermacentor marginatus, Dermacentor reticulatus, Haemaphysalis concinna, Haemaphysalis inermis, Haemaphysalis punctata, Hyalomma marginatum, Ixodes frontalis, Ixodes hexagonus, Ixodes kaiseri, Ixodes ricinus, Rhipicephalus bursa, Rhipicephalus sanguineus s.l. and Rhipicephalus turanicus. Ixodes kaiseri is for the first time recorded in the fauna of Croatia. Of the thirteen hard tick species analyzed in this study, pathogens from different groups (bacteria, protozoa and viruses) have been detected in eight species in Croatia so far. For the important vector species R. sanguineus s.s., new distributional data for Croatia are given. The standard COI barcoding region was amplified, and the sequences were analyzed by species delimitation methods together with the sequences of conspecific and congeneric species from the public BOLD database. Our specimens of H. punctata represent a new, genetically distinct MOTU. A brief overview of the available public DNA barcoding data for Ixodidae is presented, highlighting the need for an integrative approach for the clarification of the taxonomic status of problematic Ixodid taxa. The results provide a basis for the establishment of a molecular data platform for the Ixodidae of the Croatian fauna., (Copyright © 2022. Published by Elsevier GmbH.)

In a total of 1536 blood serum samples analysed by ELISA, antibodies for IAV nucleoprotein (NP) were detected in 30.3%. Results from HI show that the most common subtype of swIAV in the Croatian pig population was H1N1 (44.6%), followed by H3N2 (42.7%) and H1N2 (26.3%). Antibodies to at least one subtype were detected in 62.19% of blood serum samples. Detection of swIAV antigen was performed by IHC and detected in 8 of 28 lung samples collected post-mortem. The matrix (M) gene was detected in nine of one hundred and forty-two lung tissue samples and in seven of twenty-nine nasopharyngeal swabs. Phylogenetic analysis of amplified HA and NA gene fragments in Croatian isolates suggests the presence of swIAV H1avN1av.

West Nile virus (WNV) is an emerging zoonotic pathogen with rapid global expansion. The virus circulation is confirmed in many countries of Mediterranean Basin and Southern and Central Europe. In our study detection of specific WNV antibodies was performed in horses and cattle sera samples collected from October 2010 to April 2011. Serum samples were randomly taken from different parts of Croatia and tested by IgG and IgM ELISA. Positive serological results were confirmed by virus neutralization assay (VN-assay) and plaque reduction neutralization test (PRNT). Results showed that WNV antibodies were present in 72 out of 2098 horse sera (3.43%) and 3 of 2695 cattle sera (0.11%). The highest seroprevalence was found in Eastern Croatia in counties next to Hungarian, Serbian and Bosnia and Herzegovinian state borders. In Adriatic part of Croatia positive animals were found only in the westernmost county, near Slovenian and Italian borders. Geographic distribution and number of positive horses indicated that WNV is highly present in Croatia and spreading from East to West. However, positive horses in westernmost part of country indicate possible second origin of spreading. Location of serological positive cattle supports the hypothesis that seropositive cattle could be indicators of high WNV activity in the respective geographic regions., (Copyright © 2012 Elsevier B.V. All rights reserved.)

To investigate the degree of genetic variability of bovine leukemia virus (BLV) strains circulating in Croatia, 29 isolates from the six largest dairy farms were examined by PCR for a segment of the gp51 env gene, followed by DNA sequencing and phylogenetic analysis. The nucleotide sequences were compared with other previously characterized BLV strains from different geographical areas, comprising all seven known BLV genotypes. The Croatian sequences showed six to eight nucleotide substitutions: six silent substitutions and two amino acid changes. Four of those substitutions were within epitopes. In comparison to the sequences of other BLV genotypes, our isolates showed the closest relationship to genotype 1 isolates PL-3252 (FJ808585) and AL-148 (FJ808573) from Argentina. The degree of variation between our sequences and those of genotype 1 was 0.2- 4.6 %. In phylogenetic trees based on 400-nt and 519-nt sequences, all of the Croatian sequences clustered separately from the other sequences, revealing a new genotype.

Bovine viral diarrhoea virus (BVDV) causes a disease that has a wide range of clinical symptoms in domestic and wild ruminants. It is a major problem in cattle and causes significant economic losses in the cattle industry. The virus infects bovines of all ages and causes both immunosuppression and reproductive, respiratory and digestive disorders. Cattle infected persistently, as a continuing source of the virus and the main factor in transmission of the disease between and among herds, are the main source of BVDV and a primary factor in the epidemiology of the disease. To determine whether a BVDV infection is persistent, two samples should be taken at 3-4 week intervals and tested for the virus antigen. Animal sera, whole blood, organ and ear notch tissue samples can be used for BVDV diagnosis. In ear notch tissue, viral antigen can be detected by an antigen enzyme-linked immunosorbent assay (antigen ELISA), immunohistochemistry (IHC) and reverse-transcription polymerase chain reaction (RT-PCR). This paper describes the development and implementation of an indirect immunofluorescence (IF) method using ear notch tissue samples for diagnosis of cattle infected persistently. Results obtained by this method show that IF is a good alternative to RT-PCR and antigen ELISA and can be a quick and accurate method in diagnosis of BVDV in cattle infected persistently with this virus., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The insoluble immune complexes (ICs) were prepared under the conditions of double immunodiffusion in gel, using the suspension of the ultrasound treated PK-15 cell-line infected with porcine parvovirus (PPV) containing both viral particles and viral proteins, as well as pig or rabbit anti-PPV polyclonal immune sera. The immunodiffusion performed in an agarose gel allows only viral subunits with a molecular mass equal to or less than 1000 kDa, rather than the viral particles, to diffuse through the gel and reach the point where the immunoprecipitate is to be formed. The immunoprecipitation under the conditions of the diffusion ensures the optimal, i.e. equimolar ratio of both immunoprecipitating components, antibody/antigen in the IC. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the Western blot analyses showed the ICs were composed of two proteins, a protein in which molecular mass corresponded to the VP2 of the PPV and a protein with a molecular mass of the IgG. This suggests that the ICs are mainly composed of the VP2 antigen and IgG class antibodies. The potency of the IC-vaccines prepared in the form of a water-in-oil-in-water emulsion was compared with that of a commercially available, inactivated oil vaccine. The vaccination of gilts, 6 weeks before mating, with the IC containing allogeneic pig antibodies, resulted in the development of high and long-lasting anti-PPV antibody titres, similar to those generated by the licenced vaccine (P > 0.01). The content of the virus material administered by the IC was twice lower than that in the licenced vaccine. Neither systemic nor local reactions were observed in the gilts during the period of the trial with the IC vaccine. The number of viable piglets per litter varied between 9 and 12 and no signs of the PPV infection were detected. Rabbits were used as one of the alternative laboratory animal models accepted for the testing of the vaccine against the PPV. The rabbit humoral immune response generated by the IC containing the allogeneic antibodies were higher than that generated by the ICs containing the xenogeneic pig antibodies. It was similar to that generated by two-times higher content of the virus material administered by a commercially available vaccine. The IC-based vaccines belong to non-replicating, subunit vaccines, which are both ecologically convenient and the safest vaccines of all.

Serologic evidence of exposure to porcine parvovirus (PPV) in the wild boar (Sus scrofa) in Croatia was investigated. Serum samples from 219 wild boars captured during 2003 from 12 different locations in the Republic of Croatia were tested by using a commercial enzyme-linked immunoassay (ELISA) and a hemagglutination inhibition (HI) test. Antibodies to PPV were detected in 91 (41.6%) of tested samples and positive results were detected in wild boar from all sample locations. Adults had a significantly higher prevalence (70%) than juveniles (31%; P < 0.01). Our results indicate that wild boar populations throughout the Republic of Croatia are exposed to PPV.

In December 2001, bluetongue (BT) was confirmed serologically by the Croatian Veterinary Institute using the competitive enzyme-linked immunosorbent assay (c-ELISA). Results of the serological testing of blood samples from ruminants in the Dubrovacko-Neretvanska County are presented (3,318 sera of ruminants from 53 herds were examined). In total, 357 bovine sera (178 or 49.9% positive), 1,268 ovine sera (174 or 13.7% positive) and 1,693 caprine sera (270 or 15.9% positive) were tested. Antibodies to BT virus serotype 9 were detected in 212 of the positive sera by serum neutralisation. A preliminary light-trap survey for midges of the Culicoides genus was also performed in the Dubrovacko-Neretvanska County. Fourteen light-trap collections from seven locations were examined and yielded a total of 4,872 Culicoides of which 4,492 (92%) belonged to the Obsoletus Complex (including C. obsoletus and C. scoticus).

Na-valproat (Apilepsin, Eftil), is is a commonly prescribed medication approved for treatment of epilepsy, trigeminal neuralgy, migraine, and bipolar disorder. Although the common adverse effects associated with Na-valproat are usually benign, more serious and fatal complication like liver failure and acute pancreatitis (AP) can occur. Acute pancreatitis may take place in all ages, regardless of dose and serum levels of the medicine. An acute pancreatitis is relatively common after intravenous administration. The authors presented a case of acute opancreatitis in 13-years girl caused by Na-valproat.

During the years from 1993 to 2000, 183 strains of Pseudomonas aeruginosa were isolated from different pathological specimens originating from dogs. Antimicrobial susceptibility patterns against 10 antipseudomonal agents were obtained on 183 P. aeruginosa strains. In vitro antimicrobial susceptibility testing was performed using the disk diffusion method (Kirby-Bauer). Antimicrobial susceptibility profiles showed that among beta-lactam antibiotics, imipenem was the most active compound. Out of the 183 strains tested, 96.7% were sensitive to imipenem. Cefoperazone showed good in vitro activity against 86.9% of the tested strains. Against ceftazidime, 77.0% of strains showed sensitivity. An old penicillin, carbenicillin, gave only 71.6% sensitive strains. Sensitivity to amikacin was 87.4% and it was 83.1% to gentamicin. Pipimedic acid, a first-generation quinolone, was the least active compound of all those tested, 47.0% were resistant. The in vitro sensitivity against enrofloxacin showed that 71.0% strains were sensitive and 26.2% showed resistance. Almost all strains tested, 93.4%, were susceptible to ciprofloxacin and marbofloxacin. Besides imipenem, the quinolone antibiotics, marbofloxacin and ciprofloxacin were the most effective against P. aeruginosa strains isolated from dogs.

Enzyme linked immunosorbent assays (ELISAs) and a nested polymerase chain reaction after reverse transcription (RT-PCR) were used for the detection of the Chinese strain (C strain) of classical swine fever virus (CSFV) in blood and tissue samples of experimentally inoculated piglets. One group of 10 piglets was inoculated with C strain material from rabbits and a second one with material from infected minipig kidney (MPK) cell culture. Tested blood samples were taken on the day of inoculation as well as on days 2, 4, 6, 8, 10, 13 and 16. Samples of spleen, tonsil and brain tissue were collected from piglets on days 6, 8, 10, 13 and 16 and tested for glycoprotein E(RNS) and protein NS2-3 using commercially available ELISA kits. E(RNS) and NS2-3 were detected earlier in blood samples of piglets inoculated with the C strain propagated in a cell culture. Regardless of propagation the presence of the viral E(RNS) and NS2-3 was detected in spleen and tonsil samples simultaneously. The C strain propagated in a cell culture was found in only one brain sample, whereas, the virus propagated in rabbits was detected in 70% of the brain samples. For the detection of the CSFV RNA in blood samples, a part within the 5' non-coding region was amplified. The differences in the results gained by antigen detection in blood samples decreased when nested RT-PCR was used.

Based on the literature data, the outbreaks of new zoonoses caused by new members of the family Paramyxoviridae are briefly presented. Some characteristics of Hendra and Nipah virus, epidemiological aspects, clinical picture and pathology are described. Fruit bats are mentioned as the key to the epidemiology of Hendra virus. The virus was isolated from affected humans, horses, and from the uterine fluids of a grey-headed fruit bat (pteropus poliocephalus). New morbillivirus designated Nipah virus was isolated from pigs and humans. It caused encephalitis and pneumonia in humans.

In two separate herds of fattening calves a sudden-onset outbreak of ocular disease with profuse lacrimation occurred. The disease resembled the early stage of infectious bovine keratoconjunctivitis but after a few days the clinical signs of bronchopneumonia appeared. From conjunctival swabs Mycoplasma (M.) bovigenitalium, M. bovirhinis and infectious bovine rhinotracheitis (IBR) virus were isolated. Moraxella bovis infection was not established. In one of the herds M. bovigenitalium was also found in the pneumonic lungs of dead calves. In one herd M. bovoculi was isolated from a cow with chronic keratoconjunctivitis, housed together with affected calves. Mycoplasmas were not isolated from ocular swabs of six bulls originating from a Reproductive Centre with temporary occurrence of unilateral serous conjunctivitis resistant to antibiotic therapy.

The bovine herpesvirus 1 (BHV1) strain Za is a conventionally attenuated strain with a 2.7 kb deletion that encompasses the complete coding region for glycoprotein E (gE). This gE-negative strain was used as whole-virus antigen in an inactivated virus vaccine. Three different antigen concentrations of this vaccine were evaluated for safety and efficacy in a vaccination-challenge experiment in calves. No adverse effects were observed in any of the calves vaccinated with the gE-negative vaccines. Calves given the vaccine with the highest antigen concentration were adequately protected against challenge; clinical symptoms were virtually absent and challenge virus shedding was significantly reduced as compared with unvaccinated calves. We developed a sensitive blocking enzyme-linked immunosorbent assay (ELISA) to detect antibodies against gE. After vaccination, calves did not produce antibodies against gE, but these antibodies were detectable within 2 weeks after challenge both in vaccinated and in unvaccinated calves. These results demonstrate the efficacy of a gE-negative inactivated BHV1 vaccine and the detectability of antibodies against gE after infection. The combined use of the marker vaccine and the gE-blocking ELISA makes it possible to differentiate between vaccinated animals and infected animals. This possibility may be very useful in BHV1 control programmes.

Sera from 22 (13 wild and nine captive) European brown bears (Ursus arctos) from Croatia were tested to 18 viral and rickettsial agents. Serologic evidence of exposure was found to the following agents (number positive/number examined): Bhanja virus (3/15), Tahyna virus (3/15), West Nile virus (4/15), Naples sandfly fever virus (1/15), human adenovirus (1/22), influenza A (1/22) and B (1/22) virus, cytomegalovirus (1/22), parainfluenza virus 1 (2/22), Chlamydia psittaci (1/22), Coxiella burnetii (2/22), and canine parvovirus 2 (CPV-2) (7/22). Evidence of exposure to arboviruses was found exclusively among free-living bears. Evidence of exposure to agents usually transmitted directly was predominant among captive bears. Canine parvovirus 2 antibodies were the most frequently found antibodies and the only antibody common to both groups of bears. This may be the first report of antibodies to CPV-2 in bears.