The infection with SARS-CoV-2 virus in cats and dogs raised issue of human-to-animal transmission of SARS-CoV-2 in domestic pets in close contacts with their owners. Our study was designed to research this in the framework of Bosnia and Herzegovina. Using ELISA, AFIAS fluorescent immunoassay, RT-qPCR and WGS on Nanopore MinION platform with ARTIC Network Amplicon sequencing protocol for SARS-CoV-2, we showed that three out of thirteen dogs and one out of five cats from the households with confirmed human cases of COVID-19 in Bosnia-Herzegovina were infected with SARS-CoV-2. The high viral RNA load was detected in samples collected from a 4-year-old male Havanese (Ct = 12.52), a 6-year-old German Shepherd (Ct = 21.36) and a 9-year-old female American Staffordshire terrier (Ct = 25.74). The antibody response in dogs and one cat was observed. The viral genetic sequences from dogs were identical to the sequences detected in the owners suggesting the human-to-animal transmission of the virus. These findings, especially the low initial Ct values detected, from the public health perspective additionally stress the need for precautionary measures to protect both humans and animals.

The objective of this study was to investigate the effects of test mixture or probiotic addition to drinking water on the growth performance of broiler chickens. A total of 240 one-day-old Cobb 500 chickens were distributed into three groups with eight replicates in each (10 chickens in each replicate). The control group of chickens (C) were without treatment. The chickens in experimental group E1 were treated with the commercial probiotic Probios® and the chickens in experimental group E2 were treated with the test mixture (Lactobacillus acidophilus culture, inactivated baker’s yeast, C vitamin, lactose and glucose) prepared using the authors’ own recipe. Treatments of chickens were conducted during the first three days of life and for three days using the chickens’ vaccination drinking water. The experiment lasted for 42 days. Feed and water were offered ad libitum during the experiment. Body weight, daily feed intake, body weight gain, feed conversion ratio (FCR), carcass weight, carcass yield and European production index (EPI) were studied in this experiment. The addition of the experimental probiotic significantly increased (P<0.05) body weight gain at 21, 35 and 42 days of age, however, the probiotic Probios® improved body weight gain over the same period without any significant difference compared to the control group. FCR was significantly improved at 21 and 35 days of age in both E1 and E2 groups, but at the end of fattening the FCR was not affected. Feed consumption was not influenced by the treatments. The results obtained indicate that carcass weight significantly increased (P<0.05) in the groups of chickens treated by the test mixture or probiotic. It was concluded that addition of test mixture or probiotic improved body weight gain, feed conversion ratio, carcass weight and EPI.

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. 

During its 132-year production, the characteristics of Livno cheesechanged, first because of the transition from sheep to cow’s milk, or their mixture.Livno cheese is specific primarly due to the presence of a specific plant cover inmountain area, climatic conditions and milk of autochthonous sheep. The purposeof this work is to determine the fatty acid composition of Livno cheese, with specialreference to the contenct of bioactive components that have a positive effect onhuman health, and to tracking any changes in their content depending on thesampling or feeding period. For the production of Livno cheese, which was sampledafter 90 days of ripening at ambient conditions, sheep’s milk was used, mentioningthat the cow’s milk was added in proportion (80:20), which is commonly used in thetraditional production of this cheese. Samples were analyzed by gas chromatographyin As Vitas laboratory in Oslo Innovation Center, according to the proceduredescribed in Luna and al. (2005). A total of 24 fatty acids were determined during thethree sampling periods (July, August and September) In cheese samples, saturatedfatty acid content (SFAs) was higher in relation to monounsaturated(MUFAs) andpolyunsaturated (PUFAs) fatty acids. The fatty acid composition of the tested cheesesamples is specific, because it contains fatty acids which have been proven to havean extremely beneficial effect on human health.

The study included a total of 127 sheep milk samples from two different areas (Livno and Travnik) in summer feeding period (July, August and September). Fatty acids in milk were determined by gas chromatography (GC). The animals were marked with the appropriate number of ear tags on the basis of which we always took samples from the same animals through different periods. Fatty acids in milk were determined by gas chromatography and the following fatty acids composition: butyric acid, caproic acid, caprylic acid, capric acid, stearic acid, oleic acid, linoleic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, rumenic acid. The fatty acid content of sheep's milk in this study showed a tendency of variation, both within and between sampling areas, and characterized by its relatively high content of saturated fatty acid (SFA) during the period of harvest.