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A total of 249 serum samples from 13 wild animal species namely fallow deer (Dama dama, n = 1), roe deer (Capreolus capreolus, n = 80), red deer (Cervus elaphus, n = 22), chamois (Rupicapra rupicapra, n = 21), mouflon (Ovis musimon, n = 4), brown hare (Lepus europaeus, n = 2), nutria (Myocastor coypus, n = 1), red fox (Vulpes vulpes, n = 97), stone marten (Martes foina, n = 12), European badger (Meles meles, n = 2), golden jackal (Canis aureus, n = 2) Eurasian lynx (Lynx lynx, n = 2) and grey wolf (Canis lupus, n = 3) were analysed for the presence of antibodies against Leptospira interrogans sensu stricto. Serum samples were examined via the microscopic agglutination test for the presence of specific antibodies against Leptospira serovars Icterohaemorrhagiae, Bratislava, Pomona, Grippotyphosa, Hardjo, Sejroe, Australis, Autumnalis, Canicola, Saxkoebing and Tarassovi. Antibodies to at least one of the pathogenic serovars were detected in 77 (30.9%, CI = 25–37%) sera. The proportion of positive samples varied intraspecifically and was the biggest in large carnivores (lynx, wolf and jackal, 86%), followed by mezzo predators: stone marten (67%) and red fox (34%), and large herbivores: red deer (32%), roe deer (25%), alpine chamois (10%) and mouflon (0%). Out of the 77 positive samples, 42 samples (53.8%) had positive titres against a single serovar, while 35 (45.4%) samples had positive titres against two or more serovars. The most frequently detected antibodies were those against the serovar Icterohaemorrhagiae. The present study confirmed the presence of multiple pathogenic serovars in wildlife throughout Slovenia. It can be concluded that wild animals are reservoirs for at least some of the leptospiral serovars and are a potential source of leptospirosis for other wild and domestic animals, as well as for humans.

The repeated occurrence of anthrax in grazing animals should be a reminder of a widespread presence of Bacillus anthracis spores in the environment. Its rapid diagnosis is critical to protect public health. Here, we report a case of anthrax in cattle that was investigated using conventional and molecular methods. In 2015, six cows suddenly died within three days and the number of dead animals increased to a total of 12 within two weeks. At necropsy, anthrax was suspected. Therefore, spleen tissue samples were collected (from 6/12 animals) and laboratory tests (microscopy, cultivation, and real-time PCR) performed. The results of tissue staining for microscopy and cultivation were in congruence, while B. anthracis real-time PCR outperformed both. Spleen tissues from all six animals were real-time PCR-positive, while B. anthracis was successfully cultivated and detected by microscopy from the spleen of only three animals. Additionally, the ear tissue from another (1/12) cow tested positive by real-time PCR, supporting the suitability of ear clippings for molecular confirmation of B. anthracis. Genotyping of the isolates using multiple-locus variable-number tandem repeat analysis (MLVA) revealed a common source of infection as all three typed isolates had an indistinguishable MLVA genotype, which has not been observed previously in Europe. The results indicate that molecular testing should be selected as the first-line tool for confirming anthrax outbreaks in animals to ensure timely protection of public health.

The European Union (EU) regulates the control of cattle diseases listed in categories A and B of the European Animal Health Law (AHL). However, no strict mandatory EU regulation exists for the control of other cattle diseases that are listed in categories C, D and E. Slovenia has five control programmes (CPs) for the latter cattle diseases: bovine viral diarrhoea (BVD), infectious bovine rhinotracheitis (IBR), enzootic bovine leukosis (EBL), bluetongue and anthrax. Two (IBR and BVD) are voluntary and the others (EBL, anthrax and bluetongue) are compulsory. The three compulsory CPs are funded by the government. All the CPs are run by the government and laboratory tests are performed by the National Veterinary Institute. The rules for the CPs are laid down in Slovenian legislation. In addition, there is a national directive for the control of salmonellosis. Both BVD and IBR are endemic and have CPs based on increased biosecurity, testing and culling or vaccination, financed by the animal owners. Slovenia has been officially free of EBL since 2005 and carries out surveillance based on serological testing of a representative number of herds and inspection of carcasses at slaughter or necropsy. Vaccination is the main disease control measure for anthrax (sporadic) and bluetongue (currently perceived free—vaccination since 2017). Lack of motivation of farmers to participate in voluntary disease CPs and to implement and follow strict biosecurity measures are the most pressing issues in improving the health status of Slovenian cattle. An overview of the existing CPs and the circumstances leading to their implementation are presented.

Agriculture is an important production sector in Albania that makes a significant contribution to the gross domestic product (GDP) and employment. The livestock sector contributes more than half of the agricultural GDP. The Albanian cattle population represents 50% of the total livestock units and accounts for 85% of the national milk production, the rest being supplied by small ruminants. Cattle productivity, health and welfare are hindered by infectious diseases, some of which are also transmissible to humans (zoonosis). The aim of this manuscript is to provide an overview of the control of selected regulated and non-EU regulated cattle diseases in Albania and to highlight specific challenges for the Albanian cattle industry. The most important infectious cattle diseases in Albania for which national control and eradication strategies are in place are bovine brucellosis, bovine tuberculosis, and anthrax, which are all zoonotic. Additionally, lumpy skin disease recently emerged in the Balkan region and is currently subject to controls. Most of the available funds and European Union support are allocated to the control of EU regulated zoonotic diseases. For control of non-EU regulated cattle diseases, no funds are available resulting in the lack of national control programmes (CPs). Based on research, clinical investigations and laboratory results, several non-EU regulated cattle infectious diseases appear endemic in Albanian dairy farms. While no national CPs exist for any of them, regional initiatives are available on a voluntary basis to control infectious bovine rhinotracheitis and bovine viral diarrhea. In the voluntary CPs, there is no monitored requirement to prove disease freedom of purchased animals and to re-evaluate the herd's free status after the introduction of animals into a herd. Data on animal movements that are routinely collected could potentially be used to control the risk of purchase, but quality needs to be further improved to increase its usefulness in disease CPs. This overview aims to collate existing information on the CPs implemented in Albania and to evaluate these to highlight gaps and threats in disease control, as well as opportunities and strengths through a SWOT (Strengths, Weaknesses, Opportunities, and Threats) analysis, with the goal of providing a framework for the future implementation of animal disease control measures in Albania.

Reports on non-invasive blood sampling are limited, and there are only a few studies on using kissing bugs (Reduviidae) and medicinal leeches (Hirudo medicinalis) for hematology and biochemistry testing in various zoo animal species. The aim of this study was to evaluate the usefulness of non-invasive blood sampling with medicinal leeches for arbovirus epidemiological investigations in various animal species from one zoo collection. Medicinal leeches were manually applied on 35 animals of 11 species. Control blood samples were obtained by venipuncture of the jugular vein. Antibodies to tick-borne encephalitic virus (TBEV) were detected by using the immunoenzymatic method or an immunofluorescent assay (IFAT), depending on the animal species. One of the 35 animals (2.9%) was seropositive (Ovis aries), whereas the rest of the samples were seronegative in both methods of sampling (non-invasive by leeches vs. invasive by venipuncture). Blood sampling using medicinal leeches showed promising results. It is likely a good alternative to other more complex and invasive methods, and it can provide significant advancement in blood sampling for preventive medicine and epidemiological studies in zoo animals.

In the 1950s, infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV) disease was clinically detected and documented in cattle for the first time in Slovenia. The bovine herpes virus 1 (BoHV-1) was confirmed several times from infected herds by virus isolation on cell cultures. To keep the IC virus-free, high biosecurity measures were introduced. Before entering the IC, all calves are serologically tested and quarantined. Bulls in Slovenian insemination centres (IC) have been negative for IBR /IPV infection since 1979. From 1985 to 1991, few large-scale studies of the prevalence of IBR/IPV were carried out. In 1985, a high percentage (56.9%) of serologically positive animals were found in large state farms with Holstein Friesian cattle. Epidemiological studies in farm with bulls' mother herds were also carried out in the farms with Simmental and Brown cows. Antibodies against BoHV-1 were detected in the serum of 2.3% of Brown cattle and 3.5% of Simmental cattle. In the year 2000, 3.4% of bulk tank milk samples from 13,349 dairy farms were detected BoHV-1 antibodies positive. The highest percentage of positive animals was found in regions with an intensive grazing system (6.2% positive) and the lowest percentage in the east part of Slovenia (0.9% positive) on farms with mostly Simmental cattle. In 2006, a total 204,662 sera of cattle older than 24 months were tested for the presence of BoHV-1 antibodies and positive cattle were detected in 3.6% of tested farms. These farms kept 34,537 animals that were potential carriers of the BoHV-1. Most of the positive farms kept Holstein Friesian cattle, descendants from the state-owned farms, which were privatised or closed after 1990. In 2015, the Administration of the Republic of Slovenia for Food Safety, Veterinary and Plant Protection issued a rule that describes the conditions for granting and maintaining the status of BoHV-1 free holdings. The rule provides a voluntary control programme for breeders who want to obtain BoHV-1 free status and are willing to cover all the cost of acquiring and maintaining that status. There has been very little response from breeders.

Monitoring infectious diseases is one of the most important pillars of preventive medicine in zoos. Screening for parasitic and bacterial infections is important to keep animals and equipment safe from pathogens that may pose a risk to animal and human health. Zoos usually contain many different animal species living in proximity with people and wild animals. As an epidemiological probe, 188 animals (122 mammals, 65 birds, and one reptile) from a zoo in Slovenia were examined for selected pathogens. Antibodies to Toxoplasma gondii and Neospora caninum were detected by ELISA in 38% (46/122) and 3% (4/122) of mammals, and in 0% (0/64) and 2% (1/57) of birds, respectively, the reptile (0/1) was negative. A statistically significant difference in T. gondii prevalence was found in Carnivora compared to Cetartiodactyla and primate antibodies to Encephalitozoon cuniculi were detected by IFAT in 44% (52/118) of mammals and 20% (11/56) of birds, respectively, the reptile (0/1) was negative. Herbivores had a higher chance of being infected with E. cuniculi compared to omnivores. Antibodies to Chlamydia abortus and Coxiella burnetii were not detected in any of the 74 tested zoo animals. The sera of 39 wild rodents found in the zoo were also examined, they were negative for all three parasites. The parasite T. gondii was detected by PCR in the tissue of two mute swans (Cygnus olor), three eastern house mice (Mus musculus), one yellow-necked field mouse (Apodemus flavicollis), and one striped field mouse (A. agrarius). Positive samples were genotyped by a single multiplex PCR assay using 15 microsatellite markers, one sample from a mute swan was characterized as type II. This micro-epidemiological study offers a better understanding of pathogens in zoo animals and an understanding of the role of zoos in biosurveillance.

In the period from 2015 to 2020, an entomological survey for the presence of West Nile virus (WNV) and Usutu virus (USUV) in mosquitoes was performed in northwestern Croatia. A total of 20,363 mosquitoes were sampled in the City of Zagreb and Međimurje county, grouped in 899 pools and tested by real-time RT-PCR for WNV and USUV RNA. All pools were negative for WNV while one pool each from 2016 (Aedes albopictus), 2017 (Culex pipiens complex), 2018 (Cx. pipiens complex), and 2019 (Cx. pipiens complex), respectively, was positive for USUV. The 2018 and 2019 positive pools shared 99.31% nucleotide homology within the USUV NS5 gene and both clustered within USUV Europe 2 lineage. The next-generation sequencing of one mosquito pool (Cx. pipiens complex) collected in 2018 in Zagreb confirmed the presence of USUV and revealed several dsDNA and ssRNA viruses of insect, bacterial and mammalian origin.

We report a case of natural infection with severe acute respiratory syndrome coronavirus 2 transmitted from an owner to a pet ferret in the same household in Slovenia. The ferret had onset of gastroenteritis with severe dehydration. Whole-genome sequencing of the viruses isolated from the owner and ferret revealed a 2-nt difference.