• Porcine astrovirus viremia in apparently healthy domestic pigs. • Possible high fecal prevalence and persistence in domestic pigs in Croatia. • The vast genetic diversity of PoAstV strains on a ...relatively confined area. • The first report of PoAstV lineage 5 presence in Europe.
Astroviruses are emerging viral agents, primarily enteropathogenic in mammals, but recently have been acknowledged to have extra-intestinal implications in humans and mink. Porcine astrovirus is thought to be widely distributed and highly prevalent among pigs, nevertheless its clinical significance remains doubtful as it can be detected in diarrheic as well as in healthy pigs. Recent reports imply the immense genetic variability among porcine astrovirus strains with five distinct lineages being characterized so far. Herein, we report porcine astrovirus circulation in the blood of healthy pigs in different age categories bred on two large industrial holdings in Croatia, with viral RNA seroprevalence of 3.89%. These are the first extra-intestinal findings of astrovirus in pigs, indicating a more complex pathogenesis than previously thought. Partial polymerase sequences of serum-derived strains provisionally clustered into porcine astrovirus lineages 2 and 4, sharing high genetic identity with previously described porcine astrovirus strains. The results were supported by detecting porcine astrovirus strains in composite fecal samples, regardless of pig category or holding tested. Phylogenetic analysis of derived strains suggested the presence of porcine astrovirus lineages previously detected in pig sera with an additional highly genetically divergent lineage 5, reported for the first time in Europe. Moreover, the existence of possible sub lineages should not be excluded. The results obtained in the present study, contribute to knowledge of porcine astrovirus pathogenesis; even though it’s possible clinical significance remains unclear. High fecal prevalence accompanied with vast genetic diversity on a relatively confined area, underscores the importance of pigs as porcine astrovirus reservoirs with eventual recombination events as a possible outcome.
Canine respiratory coronavirus is a relatively new addition to the list of pathogens causing canine infectious disease complex. The virus is highly contagious, with a high prevalence in the dog ...population worldwide, especially in shelters. This study aimed to establish the presence and risk factors associated with infection in privately owned dogs and breeding colonies. This study was the first to demonstrate the presence of canine respiratory coronavirus in Croatia. Out of the 257 serum samples, 35.03% of dogs from breeding kennels and 43% of pet dogs tested enzyme-linked immunoassay positive, but the difference was not statistically significant. Sex was not an important risk factor, but the seropositivity rate increased with age. Mixing of dogs during hunting, training and dog shows was not associated with a higher seroprevalence in the breeding colonies. Daily cleaning and disinfection showed little effect on the infection spread. The study was done on a limited sample. However, it still provides evidence that the epizootiology of this disease is complex. There is no available vaccine for canine respiratory coronavirus, and further studies on environmental and risk factors will give the valuable data needed to prevent this disease. Keywords: canine respiratory coronavirus; seroprevalence; breeding colonies; ELISA; Croatia Pseci respiratorni koronavirus nedavno je dodan na listu patogena koji uzrokuju zarazni kašalj legla. Zbog svoje izrazite kontagioznosti proširio se diljem svijeta, a osobito je cest u pasa u skloništima za nezbrinute životinje. Osnovni je cilj ovog istraživanja bio ispitati proširenost infekcije u pasa u privatnom vlasništvu i uzgajivacnicama te ustanoviti cimbenike rizika koji pogoduju infekciji. Ovo je bilo prvo istraživanje koje je dokazalo prisutnost pseceg respiratornog koronavirusa u Hrvatskoj. Od 257 pretraženih uzoraka seruma 35,03% uzoraka pasa privatnih vlasnika i 43% uzoraka pasa iz uzgajivacnica dalo je pozitivan rezultat imunoenzimnog testa. Važno je napomenuti da razlika u seroprevalenciji izmedu ovih dviju populacije nije bila statisticki znakovita. Spol takoder nije bio znakovit cimbenik rizika, ali je seroprevalencija rasla s porastom dobi životinja. Miješanje životinja iz razlicitih uzgoja, prilikom lova, dresure ili izložbi pasa, nije dovelo do porasta seroprevalencije u uzgajivacnicama. Ni provodenje dnevne dezinfekcije nije znakovito utjecalo na seroprevalenciju. Ovo je istraživanje provedeno na ogranicenom broju uzoraka, ali ipak dokazuje složenost epizootiologije infekcije psecim respiratornim koronavirusom. Kako cjepivo još uvijek nije dostupno, dodatna istraživanja cimbenika okoliša i domacina, važnih za epizootiologiju ove bolesti, dat ce važne podatke nužne za provodenje mjera prevencije. Kljucne rijeci: pseci respiratorni koronavirus; seroprevalencija; uzgajivacnice; ELISA; Hrvatska
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.
The feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) may cause persistent, lifelong and lethal infections in domestic and wild felids worldwide. FIV has been confirmed in most ...Felidae species, while FeLV infection is rare among non-domestic cats. The view that retroviruses are pathogenic in domestic cats but not in other free-ranging felid species was disproved by recent findings of retroviral pathology in several wild felids. The epidemiology of retroviral infections in felids in Croatia was only investigated in urban domestic cats, while there are no data for wild cat species. As the reintroduced Dinaric lynx (Lynx lynx) population suffers from low genetic diversity, which reduces their ability to adapt to new viral outbreaks, the health status of this lynx population is of particular concern. Two different commercial immunochromatographic assays were used for qualitative detection of FIV antibodies and FeLV antigens, while PCR was used for amplification of proviral gag and env genes in Eurasian lynx blood samples. All the 17 Eurasian lynx samples collected between 2001 and 2019 tested negative in both immunochromatographic and molecular tests. Even though our sample size was rather small, considering the fact that the population size of lynx in Croatia is estimated at 40 - 60 animals, our results can be considered representative for the population’s health status. Also, data about retroviral prevalence in Eurasian lynxes are scarce, so any new findings are very valuable.
This study was performed on 29 domestic cats with a variety of clinical signs, possibly related to FIV infection. Blood samples were tested by a rapid immunochromatographic (ICA) procedure for ...detection of FIV antibodies. Subsequently, polymerase chain reaction (PCR) was performed to amplify a portion of the proviral gag gene. All 11 positive PCR products were sequenced and compared with previously reported FIV sequences. Croatian proviral isolates that could be amplified were clustered within subtype B, and additional heterogeneity was confirmed by the formation of three separate clusters. Phylogenetic analysis of circulating strains in Croatia and in southeast Europe is necessary to improve diagnostic methods and selection of the appropriate vaccinal strains.
The objectives of this study were to describe the epizootiological and clinical features of canine patients with leptospirosis, and to determine the association between the health parameters observed ...and patient survival. The study population consisted of sixty patients admitted to the Department of Microbiology and Infectious diseases with Clinic, Faculty of Veterinary Medicine in Zagreb, Croatia from 2009 to 2017. The majority of the infections were caused by serogroup Pomona (50%), while serogroups Icterohaemorhagiae, Grippotyphosa, Australis and Sejroe were identified in 30%, 8.3%, 5% and 1.7% of the patients, respectively. At initial presentation, the most frequently detected organ dysfunction was renal failure (85%), followed by pulmonary (60%) and hepatic injury (58.3%). When the frequency of animals with involvement of organ systems was calculated, 31.7% had three systems involved, 46.7% had two systems involved, 13.3% had one, and 5% did not have any system involved. The mortality risk in the group that had 0, 1, 2 and 3 systems involved was 0%, 62.5%, 46.4%, and 89.5%, respectively. In the univariable analysis, several factors were univariably associated with the risk of a lethal outcome including: day of admission (P<0.01), sex (P = 0.09), Leptospira vaccination status (P = 0.11), the presence of moderate to severe acute kidney injury (P<0.01), pulmonary involvement (P = 0.11), and several other parameters. In the multivariable Cox's model, the presence of moderate and severe kidney injury was identified as a statistically significant factor associated with lower survival. The frequency of severe clinical cases of canine leptospirosis caused by the serogroup Pomona found in this study supports the need to consider inclusion of strains of this serogroup in vaccines available on the European market. Key words: dog; lepto spiro sis; Pomona; survival
Malassezia pachydermatis is an opportunistic yeast regularly isolated from the healthy and diseased skin of various animal species. There is scarcity of data about its presence on pinnipeds' skin and ...even less information about its pathogenicity in aquatic animals. In this study, a twenty-seven year old California sea lion (Zalophus californianus) was presented with alopecia and a cutaneous lesion on the right side of the trunk that had appeared in the spring, worsened through the summer and significantly improved in the winter months. Swab samples were obtained from the lesion and healthy skin sites (tail, posterior flippers, and trunk) and M. pachydermatis was recovered from all samples. The number of colonies grown from the lesion site was significantly higher than those cultured from the healthy skin. Employing the ITS-1 molecular marker we investigated whether there is any connection between sequence types of M. pachydermatis isolates recovered from the lesion, and those isolated from the healthy skin sites. Sequencing of the ITS-1 region of all isolates revealed a 99% match to the sequence of M. pachydermatis. In addition, all sequences belonged to the same genotype and there was no difference found between the isolates obtained from the lesion site and the healthy skin. Key words: Malassezia pachydermatis; alopecia; cutaneous lesion; California sea lion (Zalophus californianus)
Feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) are among the most common infectious diseases of cats and have a global impact on the health of domestic cats. Both viruses ...belong to the Retroviridae family and like other members of this family they are associated with lifelong infection after integration of the proviral DNA into the host cell genome. Prevalence data are necessary to define the risk factors, and prophylactic, management, diagnostic and therapeutic measures for stray and owned sick cats. In this study 324 domestic cats were tested with commercially available assays. The tested cats were divided into two groups, stray and owned sick cats. The overall percentage of seropositives for FIV infection was 18.51% and the prevalence for FeLV infection was 14.50%. FIV prevalence ranged from 13.13% in stray cats up to 20.88% within the sick owned cat group. The prevalence for FeLV infection was 6.06% in stray cats and 18.22% among sick owned cats. Regarding FIV infection, our study confirmed a significantly higher percentage of seropositives for male cats, as well as for sexually intact ones within the sick owned group. Males were significantly more likely to have positive results for both retroviral infections. The study confirmed the high rate of retroviral infections in cats from the Zagreb urban area. Males, sexually intact ones, and territorial aggression are predisposing factors for FIV infection, but not for FeLV. Preventive measures should include identification and segregation of infected cats, castration of outdoor male cats, and vaccination. Key words: FIV; FeLV; prevalence; riskfactors; prevention; Croatia
BARBIC, LJ., V. SAVIC,K. KOVACEVIC, J. KAPETAN, V. STEVANOVIC, S. KOVAC, S. HADINA, Z. MILAS, V. STARESINA, N. TURK, M. PERHARIC, J. MADIC: Outbreak of equine influenza in Croatia in 2015 and post ...outbreak epidemiological situation. Vet. arhiv 88, 437-451, 2018. In March 2015, a few days after a major horse fair event in Bjelovar, Croatia, an equine influenza outbreak began and in the days that followed spread to more than 20 stud farms in the continental part of Croatia. The epidemiological investigation showed the importance of the national and international movement of asymptomatic carrier animals as a major risk factor for the introduction of the equine influenza virus and its spread in the naive population. Molecular characterization and phylogenetic analyses confirmed that the EI outbreak was caused by an imported viral strain of the H3N8 subtype, phylogenetically similar to recent European strains belonging to Florida sublineage clade 2. The post-outbreak equine influenza seroprevalence in continental Croatia, based on ELISA testing, was 12.3% and varied between 1.1% and 32.6% on a county level. The highest seroprevalence in counties with predominantly sport and leisure horses highlighted animal management as a principal risk factor for equine influenza infection. On the other hand, variations in haemagglutination inhibition titres in the tested serum samples suggested different times of infection acquisition and unreported outbreaks of equine influenza in continental Croatia after the 2015 outbreak. Vaccination coverage, even after the large outbreak in 2015, was still below 10%, which suggests the lack of education of horse owners and represents a high risk for further outbreaks. In conclusion, influenza is a wide spread infection with multiple, often unrecorded, outbreaks in continental Croatia. A high risk of further outbreaks is present due to low vaccination coverage, an increase in the sport and leisure horse population and the intensive movement of those animals. In order to prevent further outbreaks of equine influenza in Croatia it is necessary to introduce adequate surveillance and mandatory vaccination at least for the sport and leisure horse population where there is international transport of animals. Key words: equine influenza; outbreak; international transport; seroprevalence; ELISA; IHA; vaccination coverage; Croatia
Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) emerged in late 2019 and has since caused a global pandemic. Experimental studies and sporadic reports have confirmed susceptibility of ...dogs and cats to SARS‐CoV‐2 infection. However, the importance of pet animals in the epidemiology of this infection is unclear. This study reports on a first large‐scale serosurvey of SARS‐CoV‐2 infections in dogs and cats in Europe. From 26 February 2020, just one day after the first confirmed human case of SARS‐CoV‐2 infection in Croatia, to 15 June 2020, dog and cat serum samples were collected from animals admitted to three veterinary facilities in Croatia. Additionally, on 25 May 2020, a total of 122 serum samples from employees of the Faculty of Veterinary Medicine University of Zagreb were collected. Total of 656 dogs and 131 cat serum samples were tested using an in‐house microneutralisation test (MNT). Human serum samples, as well as 172 randomly selected, dog sera were tested using enzyme‐linked immunosorbent assay (ELISA). ELISA‐positive human sera were subsequently tested using MNT. Neutralising antibodies were confirmed in 0.76% cats and 0.31% dogs. ELISA reactivity was recorded in 7.56% tested dog sera. On the other hand, 5.19% of administrative, basic and pre‐clinical sciences department personnel and 5.13% of animal health service providers and laboratory personnel tested ELISA positive. Neutralising antibodies were not confirmed in any of the human samples. In conclusion, seropositivity among pet animals in Croatia is low, especially when compared to results from China. A small number of seropositive animals with a low titre of neutralising antibodies suggest infections are rare and are following infections in the human population. Additionally, contact with animals does not seem to be an occupational risk for veterinary practitioners.
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