Simple Summary Mastitis is defined as the inflammation of the mammary gland and is one of the most widespread and economically important diseases of dairy cows. Bacteria are the most reported mastitis-causative agents, while other pathogens are often overlooked because they are not routinely investigated. Incomplete diagnosis may result in inappropriate antimicrobial therapy, treatment failure, antimicrobial resistance, dissemination of pathogens, and mastitis recurrences. Thus, this study aimed to investigate the presence of not only bacteria but also other microorganisms associated with cattle mastitis on dairy farms in Bosnia and Herzegovina, a country that lacks an effective mastitis control programme and bacteriological analysis of mastitic milk. The current study revealed Mycoplasma bovis as the main pathogen and a variety of other mastitis-causing agents in cattle: bacteria (Escherichia coli, Staphylococcus aureus, coagulase negative staphylococci, Streptococcus agalactiae, Streptococcus uberis, and others), fungi (Candida spp.), and algae (Prototheca zopfii). The finding of mastitis cases requiring currently unavailable treatment and vaccines emerges in the broader scope of etiological agents in routine mastitis diagnosis. These measures applied at the herd and national levels are crucial for more effective mastitis control, animal health and welfare, the dairy industry, and public health. Abstract To obtain improved insights into the complex microbial aetiology of bovine mastitis, this study investigated the pathogens involved in cattle mastitis in Bosnia and Herzegovina. A total of 179 milk samples from cows with clinical mastitis (CM) and subclinical mastitis (SCM), as well as eight bulk tank milk (BTM) samples from 48 dairy farms, were analysed by standard bacteriological and mycological methods. Mycoplasma detection and identification were performed using culture techniques and real-time polymerase chain reaction (PCR). A total of 88 (49.2%) mastitis samples were positive for known mastitis pathogens at 32 of 47 farms (68.1%). Mycoplasma bovis was a predominant pathogen (25/187; 13.4%) in the majority of herds (14/48; 29.2%) and accounted for 48.9% of positive CM samples. Escherichia coli was the second most dominant CM pathogen (34%), followed by Streptococcus agalactiae (10.6%), whereas Staphylococcus aureus and coagulase-negative staphylococci were the most common in SCM samples (17.1%). Other mastitis pathogens included Candida spp. and Prototheca zopfii. Two BTM samples were positive for M. bovis only, and one was positive for a mixed culture of S. aureus and Streptococcus uberis. The finding of various causative agents of bovine mastitis, with M. bovis emerging as the main pathogen, emphasizes the significance of comprehensive testing that includes not only common mastitis pathogens but also mycoplasmas, fungi, and algae.

Bats are a natural host for a number of viruses, many of which are zoonotic and thus present a threat to human health. RNA viruses of the family Filoviridae, many of which cause disease in humans, have been associated with specific bat hosts. Lloviu virus is a Filovirus which has been connected to mass mortality events in Miniopterus schreibersii colonies in Spain and Hungary, and some studies have indicated its immense zoonotic potential. A die-off has been recorded among Miniopterus schreibersii in eastern Bosnia and Herzegovina for the first time, prompting the investigation to determine the causative agent. Bat carcasses were collected and subjected to pathological examination, after which the lung samples with notable histopathological changes, lung samples with no changes and guano were analyzed using metagenomic sequencing and RT-PCR. A partial Lloviu virus genome was sequenced from lung samples with histopathological changes and found to be closely related to Hungarian and Italian virus sequences. Further accumulation of mutations on the GP gene, coding the glycoprotein responsible for cell tropism and host preference, enhances the need for further characterization and monitoring of this virus to prevent spillover events and protect human health.

Mycoplasma ovipneumoniae is an important pathogen in sheep, goats, and wild ruminants. We sequenced M. ovipneumoniae strains 150 and 274 from Bosnia and Herzegovina. Strain 150 has a circular genome of 1,053,380 bp with 29.15% GC content while strain 274 has 1,081,404 bp with 28.82% GC content. ABSTRACT Mycoplasma ovipneumoniae is an important pathogen in sheep, goats, and wild ruminants. We sequenced M. ovipneumoniae strains 150 and 274 from Bosnia and Herzegovina. Strain 150 has a circular genome of 1,053,380 bp with 29.15% GC content while strain 274 has 1,081,404 bp with 28.82% GC content.

Mycoplasma ovipneumoniae, a well-established respiratory pathogen of sheep and goats, has gained increased importance recently because of its detection in wild ruminants including members of the Cervidae family. Despite its frequent isolation from apparently healthy animals, it is responsible for outbreaks of severe respiratory disease which are often linked to infections with multiple heterologous strains. Furthermore, M. ovipneumoniae is characterized by an unusually wide host range, a high degree of phenotypic, biochemical, and genomic heterogeneity, and variable and limited growth in mycoplasma media. A number of mechanisms have been proposed for its pathogenicity, including the production of hydrogen peroxide, reactive oxygen species production, and toxins. It shows wide metabolic activity in vitro, being able to utilize substrates such as glucose, pyruvate, and isopropanol; these patterns can be used to differentiate strains. Treatment of infections in the field is complicated by large variations in the susceptibility of strains to antimicrobials, with many showing high minimum inhibitory concentrations. The lack of commercially available vaccines is probably due to the high cost of developing vaccines for diseases in small ruminants not presently seen as high priority. Multiple strains found in affected sheep and goats may also hamper the development of effective vaccines. This review summarizes the current knowledge and identifies gaps in research on M. ovipneumoniae, including its epidemiology in sheep and goats, pathology and clinical presentation, infection in wild ruminants, virulence factors, metabolism, comparative genomics, genotypic variability, phenotypic variability, evolutionary mechanisms, isolation and culture, detection and identification, antimicrobial susceptibility, variations in antimicrobial susceptibility profiles, vaccines, and control.

Little is known about the presence of methicillin-resistant Staphylococcus aureus (MRSA) in a canary bird (Serinus canarius domesticus). Although MRSA in a canary bird was previously reported, to the best of our knowledge, this is the first full description of the isolation and antibiotic resistance pattern of MRSA in this pet bird. A swab was taken from the nonhealing wound on the lateral thigh of a four-year-old, caged, housed alone, male canary bird. After the identification of Staphylococcus aureus, the antibiotic susceptibility profile of the isolate was obtained by the disk diffusion test. According to the resistance to Oxacillin and Cefoxitin, the isolate was identified as MRSA. The mecA gene was confirmed by PCR. The bird was treated by offering drinking water medicated with an injectable enrofloxacin formulation at 200 mg/L over 10 days period. Two weeks after therapy, intensive contraction of the wound was observed with a reduction in size. A week later, the complete epithelization of the wound defect was verified. In this study, we could not confirm the source of infection in a canary bird, but we believe that transmission was from wild birds when the cage was putting out in order to allow the bird to sunbathe or more likely via contact with the owner. The results of this study underline the necessity for further investigations on the epidemiological role of canary birds as potential reservoirs of MRSA.

The objective of this study was to determine and compare the antimicrobial resistance profiles of clinical Staphylococcus pseudintermedius (n = 90) and Staphylococcus aureus (n = 61) isolates from different animal species in Bosnia and Herzegovina. Among S. pseudintermedius isolates from dogs (n = 86), resistance to penicillin (87.2%) was most common followed by amoxicillin (76.7%) and amoxicillin/clavulanic acid (67.4%). A total of 25 isolates were found to be resistant to oxacillin of which 21 were mecA-positive and multidrug resistant (MDR). The most frequent MDR pattern was penicillins-lincosamides-cephalosporins-macrolides-fluoroquinolones. Among the 61 methicillin-susceptible S. pseudintermedius isolates, 18 were MDR (penicillins-lincosamides-macrolides). Most of S. aureus isolates were resistant to penicillin (50.8%), followed by amoxicillin (41%) and amoxicillin/clavulanic acid (31%). Resistance to cefoxitin was detected in only two isolates. All S. aureus isolates were mecA and mecC-negative. MDR was observed in six S. aureus isolates of which five were from cattle (penicillins-lincosamides-fluoroquinolones). Isolates from red foxes (Vulpes vulpes) were sensitive to most of the antimicrobials tested. The high number of methicillin-resistant S. pseudintermedius and MDR isolates in dogs exposed in this study underlines the urgent need for establishment of national antimicrobial resistance surveillance program in animals in the country, as well as for the surveillance of veterinary antimicrobial consumption.

The objective of this study was to determine the minimum inhibitory concentrations (MICs) of nine antimicrobials (enrofloxacin, ciprofloxacin, norfloxacin, gentamicin, spectinomycin, oxytetracycline, tylosin, florfenicol, and tiamulin) against 24 Mycoplasma ovipneumoniae isolates obtained from sheep and goats and to compare the resulting antimicrobial profiles. Enrofloxacin and ciprofloxacin had the lowest MIC50 values (<0.03 μg/mL) and MIC90 values (0.25 μg/mL) for all tested isolates. The highest MIC50 value (2 μg/mL) was obtained for florfenicol, while oxytetracycline and tylosin exhibited the highest MIC90 values (16 μg/mL). The MIC values for all fluoroquinolones and oxytetracycline were significantly lower for sheep isolates. Sheep isolates were considerably more susceptible to norfloxacin and tylosin than were goat isolates. This study demonstrated differences in antimicrobial susceptibilities between sheep and goat isolates, revealing M. ovipneumoniae in goat isolates to be less susceptible. The results suggest a possible link between antimicrobial profiles of M. ovipneumoniae isolates and their host ruminant species.

Mycoplasma ovipneumoniae (Movp) is an emerging pathogen that causes respiratory disease in small ruminants worldwide. It is considered to be difficult and time consuming to grow, which complicates diagnostic and control measures including isolation (an essential step required prior to the characterisation of strains), antimicrobial susceptibility testing and the development of vaccines. The objectives of this study were to analyse in vitro growth patterns of Movp strains, and the effects of different media used to support their growth. The study was conducted on 20 ovine and caprine Movp strains, isolated using Thiaucourt's medium. The rapid growth phase varied among the strains from 24 h to 72 h, although 60% of strains (12 of 20) reached a peak at 48 h. All strains were viable at 72 h after incubation, and declining viability was observed at 96 h (13 of 20 remained viable; 65%), 120 h (9 of 20; 45%) and 144 h (4 of 20; 20%). Growth was not detected at 168 h. All strains were able to grow in modified tryptone soy broth, while PH mycoplasma medium-Hayflick modified medium supported the growth of only two strains. Improved techniques of Movp cultivation require consideration of the growth variability among strains, the time of subculturing (during the first three days of incubation) and selection of appropriate media.

Little is known about the presence of mycoplasmas in the genital tracts of domestic and stray bitches or in the vaginas of ovariohysterectomized (OHE) bitches. Moreover, to our knowledge, there has been no research to investigate the presence of canine vaginal mycoplasmas during the different stages of the reproductive cycle. We investigated the occurrence of mycoplasmas in the vaginas of healthy domestic and stray intact bitches, to correlate their presence with specific stages of the reproductive cycle, and to compare them with those in OHE bitches. We also investigated the presence of uterine mycoplasmas. Mycoplasmas were isolated from 41 of 122 vaginal swabs (34%) from domestic (27%) and stray (39%) bitches. Mycoplasma canis was the most commonly identified species (n = 26; 63%), and was detected in both intact (60%) and OHE (73%) bitches. Mycoplasma isolates from the vaginas of healthy bitches did not vary during the various stages of the estrous cycle. Mycoplasmas were not detected in uterine samples.

The objective of the present study was to investigate the presence of Aspergillus species in Bosnian sudžuk, the most preferred dry sausage in Bosnia and Herzegovina, and evaluate their aflatoxigenic potential. A total of 145 bulk and vacuum packed samples of Bosnian sudžuk were collected from retail in the Sarajevo region; 105 samples from individual artisanal producers, and 40 samples were industrially produced Bosnian sudžuk. In total, four and seven A. flavus and A. parasiticus were isolated, respectively, while one or more genes responsible for biosynthesis of aflatoxins (nor-1, ver-1, omt-1 and apa-2) were detected in eight isolates. Aflatoxigenic strains were not found in the industrially produced samples. This is the first report about the presence of aflatoxigenic Aspergillus species in Bosnian sudžuk. Key words: A. flavus; A. parasiticus; aflatoxigenicity; Bosnian sudžuk, aflatoxin genes

1) Department of Ecology, Faculty of Natural Sciences, University of Sarajevo, Zmaja od Bosne 33, 71000 Sarajevo, Bosnia and Herzegovina 2) Herpetological Association in Bosnia and Herzegovina: ATRA, Alipašina 207, 71000 Sarajevo, Bosnia and Herzegovina 3) Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium 4) Department of Microbiology, Faculty of Veterinary Medicine, University of Sarajevo, Zmaja od Bosne 90, 71000 Sarajevo, Bosnia and Herzegovina