Since the role of wild rodents/small mammals in hepatitis E virus (HEV) epidemiology has been a subject of considerable debate, this study was conducted to investigate the potential presence of HEV ...RNA in small rodents collected within their natural habitats and to detect if they can be potential reservoirs of the virus. A total of 483 small rodents were captured using snap traps placed at 11 regions in Croatia. Sampling was undertaken in 2008 and repeated from 2010 to 2014. Liver samples were tested for the presence of HEV RNA. HEV RNA was detected in only one liver sample (0.21%) originated from Apodemus flavicollis from the location Medvednica, nearby Zagreb collected in 2014. According to the sequence analysis, the isolate has shown to be a member of Orthohepevirus A species, genotype HEV-3. The genotyping results confirmed grouping into subtype 3a, general cluster 3abchij.The detected HEV strain showed to be genetically highly related to strains found in humans and/or domestic pigs and wild boars from Croatia. Our finding indicates that wild small mammals could play a role in the epidemiology of HEV-3 infection and therefore should be taken under consideration as potential reservoirs or/and transmitters of the disease. However, further investigation is needed to recognize their potential for maintaining the infection in natural conditions.
Female as well as male mate choice has already been recognized in many insects. The traits responsible for the mate choice in a striped stink bug (
Graphosoma lineatum
L.) are not yet explored. This ...works concentrates on few possible traits responsible for mate choice in a striped stink bug. For this analysis free living copulating individuals as well as the single ones were collected. On two different locations animals were collected and their body length, pronotum broadness and coloration were measured. No differences were found in all measured traits between copulating and control individuals, showing no preferences when choosing copulation partner. Results also show no evidence of assortative mating, referring to the body length and the pronotum broadness of both sexes. All three measured traits seem not to have an influence on the mate choice of both sexes. Further experimental testing is needed to fully excluded importance of body size and coloration in mate choice of a striped stink bug.
Odabir partnera od strane ženke, kao i mužjaka, već je prepoznat kod mnogih kukaca. Evoluirale su različite strategije odabira partnera na temelju veličine tijela i zabilježene su kod mnogih vrsta kukaca, uključujući i neke smrdibube (Hemiptera: Pentatomidae). Osim veličine tijela kod kukaca je zabilježen i utjecaj obojenosti na odabir partnera. Osobine odgovorne za odabir partnera prugastog smrdljivca (
Graphosoma lineatum
L.) nisu detaljno istražene. Ovaj se rad usredotočuje na utjecaj veličine tijela i obojenosti oba spola kod odabira partnera prugastog smrdljivca. Ukupno 10 dana su na dvije lokacije prikupljane kopulirajuće jedinke, kao i slobodne (nekopulirajuće) jedinke. Životinjama je determiniran spol, izmjerena duljina tijela, širina pronotuma i zabilježena boja. Jedinke u kopulaciji su brojevno označene u svrhu izuzimanja iz analize podatke ponovljenih parenja. Podaci uključuju ukupno 65 različitih kopulacijskih kombinacija, 44 jedinki kontrolnih ženki i 53 kontrolna mužjaka. Za sve tri mjerene osobine nisu pronađene statistički značajne razlike između kopulirajućih i kontrolnih (nekopulirajućih) jedinki, što ukazuje da ne utječu na preferencije prilikom odabira partnera. Rezultati također ne pokazuju dokaze asortativnog parenja, barem što se tiče duljine tijela i širine pronotuma. Kod oba spola čini se da sve tri mjerene osobine nemaju utjecaja na izbor partnera. Unatoč dobivenom, potrebna su daljnja ispitivanja kako bi se u potpunosti isključila važnost veličine tijela i boje u odabiru partnera prugastog smrdljivca, kao i ispitivanja vezana uz mogući utjecaj kemijskih i vibracijskih signala.
Tick-borne diseases are a serious threat to both public and veterinary health. In this study, we used high-throughput sequencing to characterize the virome of three tick species implicated in the ...spread of vector-borne disease throughout Croatia. Ten viruses were identified, including seven potential novel species within the viral families
,
,
,
,
, and
.
Croatia is a focus for many rodent-borne zoonosis. Here, we report a survey of 242 rodents and small mammals, including 43 Myodes glareolus, 131 Apodemus flavicollis, 53 Apodemus agrarius, three ...Apodemus sylvaticus, six Sorex araneus, four Microtus arvalis, one Microtus agrestis, and one Muscardinus avellanarius, collected at eight sites in Croatia over an 8-year period. Multiplex MassTag polymerase chain reaction (PCR) was used for detection of Borrelia, Rickettsia, Bartonella, Babesia, Ehrlichia, Anaplasma, Francisella tularensis, and Coxiella burnetii. Individual PCR assays were used for detection of Leptospira, lymphocytic choriomeningitis virus, orthopoxviruses, flaviviruses, hantaviruses, and Toxoplasma gondii. Of the rodents, 52 (21.5%) were infected with Leptospira, 9 (3.7%) with Borrelia miyamotoi, 5 (2%) with Borrelia afzelii, 29 (12.0%) with Bartonella, 8 (3.3%) with Babesia microti, 2 (0.8%) with Ehrlichia, 4 (1.7%) with Anaplasma, 2 (0.8%) with F. tularensis, 43 (17.8%) with hantaviruses, and 1 (0.4%) with an orthopoxvirus. Other agents were not detected. Multiple infections were found in 32 rodents (13.2%): dual infections in 26 rodents (10.7%), triple infections in four rodents (2.9%), and quadruple infections in two rodents (0.8%). Our findings indicate that rodents in Croatia harbor a wide range of bacteria and viruses that are pathogenic to humans.
Orthohantaviruses are mainly carried and transmitted by wild rodents, although during the last decade, they have also been identified in multiple species of shrews and moles. Orthohantavirus,
(Seewis ...virus, SWSV), first detected in Switzerland in a single
(Eurasian common shrew) specimen, has been further described in several European countries, including Croatia's neighboring Slovenia and Hungary. Croatia is a well-known endemic region for several zoonotic agents including three different orthohantaviruses:
(PUUV),
(DOBV), and
(TULV). In this study, nine shrews were tested and SWSV RNA was detected in liver, lung, and kidney belonging to two shrews (22.22%), one collected on Medvednica mountain in Zagreb County, and the other in the Stara Gradiška area in lowland Croatia. The phylogenetic analysis of the complete S segment's open reading frame (ORF) and partial L-segment revealed that the Croatian sequences, when compared to sequences from the adjacent geographic regions, form a specific genetic lineage. Two SWSV-positive shrew species-
and
(Mediterranean water shrew)-were identified using barcode-based sequence analysis. Therefore, the SWSV detection in
throughout the course of this study is seen as a rare find in this shrew species. To our knowledge, this is the first molecular and phylogenetic analysis of SWSV in Croatia.
Altitudinal distributions, population structures and seasonal dynamics of tick fauna at three localities in Continental Croatia (Medvednica and Papuk) and an alpine biogeographic region (Gorski ...Kotar) were studied. Sampling of questing ticks was performed twice a year (spring and autumn, from 2019 to 2021) at different altitudes (200, 400, 600, 800 and 1000 m above sea level) using the flagging method. In total, 2942 ticks (53.9% larvae, 40.1% nymphs, 6.0% adults) were sampled and 2937 (99.83%) were determined as Ixodes ricinus, 4 (0.14%) as Haemaphysalis concinna and 1 (0.03%) as Ixodes frontalis. Ixodes ricinus was the only species found at all altitudes and sampling sites. The highest tick abundance was recorded at higher altitudes (800–1000 m asl.) on Medvednica and in Gorski Kotar within mixed forests of European beech and European silver fir, while on Papuk most of the ticks were sampled at lower altitudes (200 m asl.) in Sessile oak forest. From 27 pools containing 305 ticks, 1 (3.7%) was positive for Borrelia burgdorferi sensu lato infection. Borrelia burgdorferi s.l. was detected in 20% (1/5) of the pools containing nymphs and adults collected in Gorski Kotar (600 and 800 m asl.). Sequencing of the ospA gene and phylogenetic analysis revealed the presence of the Borrelia burgdorferi sensu stricto genotype.
Small rodents (Rodentia, subfam. Murinae: real mice, Arvicoline voles) greatly affect natural regeneration, stability and dynamics of forest communities worldwide. Every 3–4 years rodent damage in ...Croatian state forests is the most severe in forest regeneration stands, especially in pedunculate oak (Quercus robur L.) and narrow-leaved ash (Fraxinus angustifolia Vahl.) forests, where rodents can seriously impede natural regeneration by damaging seeds, stems and roots of saplings. These negative interactions are an even bigger challenge nowdays as pedunculate oak and narrow-leaved ash have become more vunerable in the last decades and are known as the most sensitive species of lowland forests in Croatia due to microclimatic and macroclimatic changes and the unfavourable interaction of a whole series of anthropogenic, abiotic and biotic factors. In the last 40 years, in Croatian state forests, rodent management consisted of monitoring and mainly rodenticide use. Trying to implement IPM (Integrated Pest Management) postulates into practice over the years, different prevention methods against small rodents were tested, but not many came to use. The aim of this research was to look into different logging residue management approaches and their effect on the rodent damage in two pedunculate oak forest regeneration stands in central Posavina in Croatia. Rodent damage on stem and root of tree saplings was recorded by visual inspection on three plots (5x5 m) with scattered logging residue, and one plot (5x5 m) with no residue at one micro-depression site (95 m a.s.l.), and on one micro-elevation (99 m a.s.l.) site. Plots with scattered logging residue represented a type of forest residue management in which logging debris (branches) is cut to smaller lengths and distributed evenly at the forest regeneration stand. Plot with no logging debris represented a residue management method in which wood mass is completely removed from the regeneration stand after felling. We counted, determined and inspected tree saplings found at chosen plots for rodent damage (on stem and roots) and also determined the average weight and moisture content of logging residue (branches around 5–7 cm in diameter) found at the site. In spring 2017, 3380 tree saplings (2978; 81% pedunculate oak, 7; 0.2% narrow-leaved ash and 395; 11.7% other deciduous species) were inspected for rodent damage. At micro-depression site, on a plot with no logging residue, only 13.4% of the saplings were damaged, while the average share of damaged saplings on three plots with scattered residue was more than six times higher; 87.8%. The average mass of the logging residue weighed at site with scattered residue was 10.14 kg kg/m2 and moisture content was 19.2%. At micro-elevation site, 25.4% of the saplings were damaged on a plot with no logging residue, while the average share of damaged saplings on three plots with scattered residue was two times higher; 51.4%. The average mass of the logging residue weighed at SRP 1–3 was 5.1 kg/m2. We also determined moderately strong positive correlation (R=0.69133) between the mass of logging residue and rodent damage and strong negative correlation (R=–0.89598) between wood moisture content of the logging residue and rodent damage. In years ahead, with unpredictable climate effects and potentially very variable small rodent dynamics, removing the logging residue after the felling could represent a residue management that contributes to a more effective and ecologically based rodent management. It could also become a usable preventive method within IPM and help prevent sever rodent damage, even during the outbreaks in pedunculate oak regeneration stands.
Babesia divergens and B. divergens-like organisms are the main causative agents of human babesiosis in Europe. Recently, the first case of human infection with Babesia microti was confirmed in ...Germany, implicating the presence of zoonotic isolates. To estimate the presence of zoonotic B. microti in Croatia we analyzed 120 small wild mammals that serve as its reservoir by polymerase chain reaction. Yellow-necked mice (Apodemus flavicollis) and bank voles (Myodes glareolus) were both found to be infected with prevalence of 16.2%. Sequence analysis of the portion of 18S rDNA gene demonstrated that six polymerase chain reaction-positive samples, detected in both rodent species, were identical to that of the human Jena/Germany strain (EF413181). The other two isolates were identical to the nonzoonotic Munich strain (AB071177). The results of this study indicate the presence of zoonotic B. microti in A. flavicollis and M. glareolus in Croatia and a potential risk for human health.
Hantaviruses, Leptospira spp., and Babesia spp. are rodent-borne pathogens present worldwide. We studied multiple co-infections of small rodents in Croatia with all three pathogens. Twenty-eight ...Apodemus flavicollis and 16 Myodes glareolus were tested for the presence of hantavirus RNA by real-time RT-PCR, Leptospira strains by renoculture method and Babesia DNA by PCR. Anti-hantavirus antibodies and anti-Leptospira antibodies were detected by serological methods. Very high infection rates with each pathogen were found in A. flavicollis: 20 of 28 rodents (71%) were infected with Dobrava virus, 13 rodents (46%) were infected with Leptospira, and 5 rodents (18%) were infected with Babesia. Multiple co-infections with all three pathogens were found in 3 of 28 (11%) A. flavicollis animals, suggesting that the same rodent host can be infected with several pathogens at the same time. Dual infections with both hantaviruses and Leptospira were found in 7 of 44 rodents (16%), with hantaviruses and Babesia in 2 rodents (5%), and double infection with both Leptospira and Babesia were found in 1 rodent (2%). Since hantaviruses, Leptospira, and Babesia have similar geographical distributions, it is to be expected that in other parts of the world multiple co-infections, representing a serious threat to public health, can be found.
In Croatia, several rodent- and vector-borne agents are endemic and of medical importance. In this study, we investigated hantaviruses and, for the first time, tick-borne encephalitis virus (TBEV) ...and Rickettsia spp. in small wild rodents from two different sites (mountainous and lowland region) in Croatia. In total, 194 transudate and tissue samples from 170 rodents (A. flavicollis, n=115; A. agrarius, n=2; Myodes glareolus, n=53) were tested for antibodies by indirect immunofluorescence assays (IIFT) and for nucleic acids by conventional (hantaviruses) and real-time RT-/PCRs (TBEV and Rickettsia spp.). A total of 25.5% (24/94) of the rodents from the mountainous area revealed specific antibodies against hantaviruses. In all, 21.3% (20/94) of the samples from the mountainous area and 29.0% (9/31) from the lowland area yielded positive results for either Puumala virus (PUUV) or Dobrava-Belgrade virus (DOBV) using a conventional RT-PCR. All processed samples (n=194) were negative for TBEV by IIFT or real-time RT-PCR. Serological evidence of rickettsial infection was detected in 4.3% (4/94) rodents from the mountainous region. Another 3.2% (3/94) rodents were positive for Rickettsia spp. by real-time PCR. None of the rodents (n=76) from the lowland area were positive for Rickettsia spp. by real-time PCR. Dual infection of PUUV and Rickettsia spp. was found in one M. glareolus from the mountainous area by RT-PCR and real-time PCR, respectively. To our knowledge, this is the first detection of Rickettsia spp. in small rodents from Croatia. Phylogenetic analyses of S- and M-segment sequences obtained from the two study sites revealed well-supported subgroups in Croatian PUUV and DOBV. Although somewhat limited, our data showed occurrence and prevalence of PUUV, DOBV, and rickettsiae in Croatia. Further studies are warranted to confirm these data and to determine the Rickettsia species present in rodents in these areas.