Background: According to the WHO (2019), more than 1.5 billion people worldwide are infected with soil-transmitted parasites. Previous research in the Federation of Bosnia and Herzegovina (FB&H) was mainly conducted in the area of the Sarajevo Canton. Therefore, the aim of the research was to explore contamination of soil and vegetation with developmental forms of parasites in the other cantons of FB&H. Methods: Between Apr and Oct 2018, a total of 1,618 soil and vegetation samples were taken from 386 different locations in the 9 cantons of the FB&H. Results: Positive samples were observed, 65/66 (98.48%) municipalities/cities and on 239/386 (61.92%) locations. Out of 1,618 samples taken in total (1,263 soil samples and 355 vegetation samples), 357 (22.06%) were positive, out of which 337 (26.68%) and 20 (5.63%) were soil and plant samples, respectively. In total, the following adult and developmental forms were identified: Taeniidae eggs (7.30%), Toxocara spp. eggs (62.08%), Ancylostomatidae eggs (25.00%), Trichuris spp. eggs (9.55%), Capillaria spp. eggs (3.37%), Toxascaris leonina eggs (1.40%), Nematodes larvae (19.38%), Giardia duodenalis cysts (5.06%), Cryptosporidium spp. oocysts (1.4%), oocysts and cysts of different species of Protozoa (3.93%). Conclusion: The identified developmental forms of parasites pose a permanent threat to human health. It is necessary to carry out measures to reduce the contamination of soil and vegetation in coordination with systematic solutions (legislation), paralelly with contribution of animal owners, veterinarians, physicians, ecologists, parents and all the others involved in this issue.

Rift Valley fever (RVF) is an arboviral zoonosis that primarily affects ruminants but can also cause illness in humans. The increasing impact of RVF in Africa and Middle East and the risk of expansion to other areas such as Europe, where competent mosquitos are already established, require the implementation of efficient surveillance programs in animal populations. For that, it is pivotal to regularly assess the performance of existing diagnostic tests and to evaluate the capacity of veterinary labs of endemic and non-endemic countries to detect the infection in an accurate and timely manner. In this context, the animal virology network of the MediLabSecure project organized between October 2016 and March 2017 an external quality assessment (EQA) to evaluate the RVF diagnostic capacities of beneficiary veterinary labs. This EQA was conceived as the last step of a training curriculum that included 2 diagnostic workshops that were organized by INIA-CISA (Spain) in 2015 and 2016. Seventeen veterinary diagnostic labs from 17 countries in the Mediterranean and Black Sea regions participated in this EQA. The exercise consisted of two panels of samples for molecular and serological detection of the virus. The laboratories were also provided with positive controls and all the kits and reagents necessary to perform the recommended diagnostic techniques. All the labs were able to apply the different protocols and to provide the results on time. The performance was good in the molecular panel with 70.6% of participants reporting 100% correct results, and excellent in the serological panel with 100% correct results reported by 94.1% of the labs. This EQA provided a good overview of the RVFV diagnostic capacities of the involved labs and demonstrated that most of them were able to correctly identify the virus genome and antibodies in different animal samples.

Whole Genome Sequence of four samples from COVID-19 outbreaks was done in two laboratories in Bosnia and Herzegovina (Veterinary Faculty Sarajevo and Alea Genetic Center). All four BiH sequences cluster mainly with European ones (Italy, Austria, France, Sweden, Cyprus, England). The constructed phylogenetic tree indicates probable multiple independent introduction events. The success of future containment measures concernig new introductions will be highly challenging for country due to the significant proportion of BH population living abroad.

Background The production of milk and dairy products and their placement on the market represent a constant profit for the farmers/producers in Bosnia and Herzegovina (BH). The profitable operation of the dairy farms is influenced by the reproductive performance of the lactating animals. This study assessed individual animal reproductive characteristics in selected dairy farms and described their reproductive performance indicators. Results The median age at first insemination was 493 days (5th–95th percentile range 429–840), while the age at first calving was 802 days (5th–95th percentile range 708–1168). The median pregnancy proportion at first insemination was 40% (5th–95th percentile range 17–62), while the cumulative pregnancy rate calculated at day-60, day-80, day-100, and day-120 showed that approximately 64% of all pregnancies happened before day-120. The calculated interservice intervals showed that approximately 69% of the repeat breeding animals came back to the oestrus in the period of 18 to 24 days. This is an indication of very good oestrus detection in selected dairy farms. The mean number of services per pregnancy was 2.61 (range 1–12). The median calving-to-first-insemination interval was 62.5 days (5th–95th percentile range 16–408). The calving-to-conception interval was 101 day (5th–95th percentile range 36–506). Finally, the calving interval was 385 days (5th–95th percentile range 329–773). Conclusions There is a need for an organised, regular, and more comprehensive recording system for the reproduction of dairy cattle among dairy farms in Una-Sana Canton. The calculated reproductive measures indicated an undulant trend in reproductive performance among selected dairy farms in Una-Sana Canton. Knowing the apparent reproductive indicators described in this study, the farmers and veterinary authorities may identify and correct areas in their management that contribute to the reproductive underperformance.

ABSTRACT Background: Towards preparation for a possible influenza pandemic, investigation of the molecular characteristics of the circulating avian H5N1 influenza virus strains is of crucial importance. These H5N1 viruses continue to spread, to infect animals and humans and to evolve and diversify providing so an ever-looming pandemic threat.Aim: To identify genetic structure and molecular biological characteristics of BiH's isolates of H5N1 HPAI as well as to assess the level of pathogenicity, phylogenetic origin and host- specificity of the isolates.Material and Methods: SPF embryonated chicken eggs were used for virus isolation. Viral RNA extracted using QIAamp viral RNA kit and manufacturer’s protocol (QIAGEN®) was used for PCR amplification. cDNA synthesis and PCR amplification of the coding region, using gene specific primer sets (primer sequences available on request), were carried out for all eight viral RNA segments separately. The Prism Big Dye Terminator v1.1 cycle sequencing kit (Applied Biosystems) was used and products were analyzed on an automatic ABI PRISM 3130 genetic analyzer (Applied Biosystems). Nucleotide sequences were analyzed using Bioedit software (v. 7.0.9.0) with an engine based on the ClustalW 1.4 algorithm. MEGA software (v. 4,0), using the neighbor joining tree inference analysis with the Tamura-Nei γ-model, was used to estimate phylogenies and calculate bootstrap values from the nucleotide sequences.Results: Full-length nucleotide sequences of the A/Cygnus olor/BIH/1/2006 (H5N1) strain were deposited in EMBL Nucleotide Sequence Database under accession nos. FN186008 to FN186014 and FM20943. The pathogenicity and host specificity of this strain, as polygenic traits, are determined in silico by the structure of its proteins, especially surface glycoproteins, HA and NA. Multibasic amino acid stretch PQGERRRKKR/GLF, marker of strains highly pathogenic to poultry, was present at the HA cleavage site of BiH strain. The RBS was typical for avian influenza viruses and contained Gln and Gly at positions 238 and 240 (H5 numbering) that is,226 and 228 according to H3 numbering with seven potential glycosylated sites but with increased binding to alpha2-6 sialoglycans thanks to substitutions, as follows, 110N, 171N, 171N, 172A, 205R and 251P. NA structure assigned this strain to the Z genotype, characterized also by the deletion of the five amino acid residues of the NS1 protein (positions 80-84). Amino acid residues, typical for the avian influenza viruses, were revealed in 40 out of 43 positions of M1, M2, NP, PA, PB2 and HA, determining the host range specificity. Phylogenetic analysis of the HA gene revealed that BiH isolates belonged to genetic clade 2.2., and presence of aspartic acid at the position of 403 of HA locate BiH isolates in 2.2.2. sub-clade.Conclusions: The BiH’s isolates were determined as HPAI virus with genes sequences closely related to A/Cygnus olor/Astrakhan/Ast05-2-10/2005 (H5N1). Three residues (M2 - 28V and 78K, NP - 33I), typical of human influenza viruses, were found, indicating a certain degree of intercurrent evolutionary adaptive changes in BiH isolates. Sequence comparison of HA and NA segments with relevant sequences in GenBank revealed that the BiH isolates and the ones from the southern Russia (Astrakhan region) group together phylogenetically, forming a monophyleticcluster in both genes indicating that these isolates have evolved from the same origin. Sequence derived phenotype markers of NA protein (E99, V129, D131, R136, H255 and Y256) as well as of M2 protein (26L, 27V, 30A, S31 and G34) showed that the isolates have an oseltamivir and amantadine sensitive genotype. 

Species identification in food has become a prominent issue in recent years as the importance of consumer protection has increased. DNA-based species identification methods were developed by researchers in the last two decades, as these are reliable, accurate, and low-cost techniques for species identification in raw and processed food products as well. In our study, universal primers were designed to conserved regions of mitochondrial 12S rRNA. Amplicons were heat-denatured and a PCR single strand conformation polymorphism (SSCP) method was developed to identify cattle, buffalo, sheep, and goat DNA. Sensitivity of this technique was tested on DNA mixtures of cattle-sheep, cattle-goat, and cattle-buffalo and the threshold limit of cattle DNA was 5%, 5%, and 3%, respectively. One hundred and five cheeses were purchased and collected from Bosnian and Hungarian farmers, retails, and supermarkets to reveal fraud, 32 percent of them (34 cheeses) were found to be mislabelled by species.

________________________________________________________________________________________ KRKALIC, L., E. SATROVIC, T. GOLETIC, P. DŽAJA, K. SEVERIN: Chlamydophila abortus infection in a fl ock of goats in Bosnia and Herzegovinaa case report. Vet. arhiv 85, 359-368, 2015. ABSTRACT The aim of this research was to determine the presence of Chlamydophila abortus (C. abortus) infection in one fl ock of goats with a previously recorded history of reproductive failures (abortion, stillbirths, weak born kids) and long-term poor reproductive performances. The affected fl ock was from the southern region of Bosnia and Herzegovina (B&H) and consisted of 48 goats kept in semi-intensive conditions. Blood samples and vaginal swabs were collected twice during 2012 and 2013 and the sample size was estimated at a 95 % confi dence level, with predicted prevalence of 20 %, using the recommendations for determining the required sample size necessary to detect the presence of disease in a fl ock. A representative sample from this fl ock was taken by simple random sampling. In the total of 12 blood sera that were tested for the specifi c antibodies against C. abortus, with the use of enzyme-linked immunosorbent assay (CHEKIT® Chlamydophila abortus Antibody Test Kit), the results showed that 11 (91.7 %) sera were positive for C. abortus antibodies. Vaginal swabs from all animals were analysed by a modifi ed Chlamydiaceae-specifi c rtPCR test, targeting the 23S rRNA gene, to determine the presence of known Chlamydiaceae, and three (25 %) samples were positive. These positive samples were subsequently tested with a test targeting the ompA gene region (ompA-rtPCR) specifi c for Chlamydophila abortus. All three samples were also positive using this test.

A successful and harmless method for rehabilitation of hygienic status of water and its supply system using a stabilized liquid chlorine dioxide solution on a farm of the laying hens affected by severe colisepticemia is described. Source of infection was drinking water contaminated by slurry from two pig facilities located above the water tank. The contaminated water caused the emergence of biofilm consisting mainly of coliform bacteria on the interior surfaces of the plastic pipes. Through drinking the contaminated water the infection of the laying flocks occurred. With the aim of improving the flocks’ health status, a programme of sanitary treatment of external and internal water supply system and water was created and implemented. In order to prevent biofilm formation and improve sanitation prescribed was the use of stabilized liquid chlorine dioxide (ClO2) in the 4‰ concentration for so-called night "shock" treatments, and 2‰ concentration for prophylactic daily disinfection of drinking water. With the improvement of the flocks’ health status, the "shock" treatments with ClO2 were repeated in the upcoming months. As an add-on therapy, 40 mg per bird of vitamin C through drinking water for three days was prescribed. The use of non-resorptive antibiotics, AD3E vitamins and amino acid supplements was excluded because they had failed to improve the flocks’ health status in the acute phase. Therefore, the sanitation programme based on the use of stabilized liquid ClO2 in the water supply system of the laying flocks affected by severe colisepticemia resulted in radical decrease of mortality during the next three months. Key words: chlorine dioxide, biofilm, sanitation, disinfection, colisepticemia

The conducted research gives an overview of the results obtained after the application of 1‰ solution of stabilized liquid chlorine dioxide on some food-born related bacteria - E. coli, Staphylococcus aureus, S. Enteritidis and C. jejuni.  For this purpose,  reference strains of the aforementioned pathogens in decimal dilutions were exposed to 1 ml of 1‰ solution of stabilized liquid chlorine dioxide for one hour. Reduction of bacteria counts per mililitre (CFU/ml) has been noticed for all bacteria, with total reduction of C. jejuni and Staphylococcus aureus in the fourth (1:104), and for S. Enteritidis and E. coli in the sixth (1:106) decimal dilution. Key words: chlorine dioxide, E. coli, S. aureus, S. Enteritidis, C. jejuni