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Highlights • Bartonella DNA was detected in 10.5%–35.9% cervids and 85% deer keds. • Two Bartonella lineages in Norwegian cervids and deer keds were detected. • One lineage of Bartonella sp. is transmitted by L. cervi. • For other lineage the possible vectors still remain unknown. Abstract Bartonella bacteria are arthropod-borne and can cause long-term bacteremia in humans and animals. The predominant arthropod vectors and the mode of transmission for many novel Bartonella species remain elusive or essentially unstudied. The aim of this study was to investigate the prevalence of Bartonella spp. in Norwegian cervids and deer keds (Lipoptena cervi) and to characterise the bacteria by sequencing of the partial gltA gene and 16 S–23 S rRNA intergenic spacer region (ITS) in order to evaluate a possible transmission route. A total of 260 spleen samples and 118 deer keds were collected from cervids by hunters in the Southern part of Norway. Bartonella DNA was detected in 10.5% of spleen samples of roe deer (n = 67), in 35.1% red deer (n = 37), in 35.9% moose (n = 156), and in 85% pools of adult wingless deer ked (n = 59). Two Bartonella lineages were identified based on phylogenetic analysis of the gltA gene and ITS region sequences. [ABSTRACT FROM AUTHOR]

Birds are increasingly considered important in the global dispersal of tick-borne pathogens. The purpose of this study was to investigate the prevalence of Borrelia burgdorferi sensu lato genospecies in Ixodes ricinus ticks collected on passerine birds from two ornithological stations (Jomfruland and Lista) in southern Norway. In both sampling sites a total of 608 (109 larvae and 499 nymphs) of immature ticks were collected from 157 infested birds representing 26 species. The presence of B. burgdorferi s.l. DNA in ticks was tested using direct PCR amplification of the ospA gene, localized on lp-54 plasmid in B. burgdorferi s.l. genome. Twenty one out of 157 infested (13%) birds carried B. burgdorferi s.l. infected Ixodes ricinus ticks. B. burgdorferi s.l. DNA was detected in 19.7% of the tick samples. In total, 6.4% of larvae (7/109) and 10% of nymphs (50/499) were positive. The vast majority of infected ticks were found on blackbird (Turdus merula). The most frequent genospecies was B. garinii (85.5%), followed by B. afzelii (9.0%) and B. valaisiana (5.5%). Five different B. garinii genotypes were identified. Our findings showed that the blackbirds may support the circulation and spread of B. garinii and B. valaisiana. B. garinii was not detected in questing ticks from investigated locations. B. afzelii genospecies, however, dominated in questing ticks, rodents and ticks from rodents. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum is a tick-borne bacterium that infects a wide range of animal species. The aim of our study was to investigate the prevalence of A. phagocytophilum in Norwegian moose Alces alces and to characterize the bacteria by sequencing of partial msp 4 and 16S rRNA genes. Hunters collected spleen samples from 99 moose of different ages during 2013 and 2014 in two areas: Aust-Agder County (n = 70) where Ixodes ricinus ticks are abundant and Oppland County (n = 29) where ticks were either absent, or abundance very low. A. phagocytophilum was detected only in moose from the I. ricinus - abundant area. The overall prevalence of infection according to 16S rRNA and msp4 gene-based PCR was 41.4% and 31.4% respectively. Sequence analysis of the partial 16S rRNA and msp4 gene revealed two and eight different sequence types respectively. Four of eight msp4 sequence types determined in this study were unique, while others were identical to sequences derived from other ruminants and ticks. The present study indicates that moose could be a potential wildlife reservoir of A. phagocytophilum in Norway. [ABSTRACT FROM AUTHOR]

Norway represents the northern limit in the geographical distribution of Ixodes ricinus ticks in Europe. During the last decade an expansion of the range of this tick further north and at higher altitudes has been detected. This could affect the spread of Borrelia burgdorferi sensu lato pathogens in new territories and increase the risk of human infection in the country. The aim of the present study was to determine the prevalence of B. burgdorferi s.l. in I. ricinus ticks in different localities of Norway. Ticks were collected in 24 locations from northwest to southeast Norway during spring-summer 2004 and 2006-2008. We used fla gene as a target for amplification of spirochete DNA. Multiplex PCR based on ospA gene of B. burgdorferi s.l. was used for species identification. The heterogeneity of B. garinii strains was investigated using ospA-based sequencing analysis. B. burgdorferi s.l. was detected in 171 (7.2%) out of 2360 I. ricinus ticks analyzed. The infection rate varied from zero to 32% in the investigated locations. Infection rate in adults (11.9%) was significantly higher than in nymphs (4.7%). The overall infection rate in ticks from western Norway (4.0%) was significantly lower than in those from southern Norway (10%). Four B. burgdorferi s.l. genospecies were identified, and the most common was B. afzelii (71.9.0%), followed by B. garinii (12.9%), B. burgdorferi s.s. (7%) and B. valaisiana (1.2%). A total of eight (4.8%) of the infected ticks harboured mixed infections. Phylogenetic relationship between B. garinii strains and their correspondence to OspA serotype types was compared with the sequences registered in GenBank database. Five genotypes of B. garinii derived from ticks were identified. The most frequent was OspA type 6. [ABSTRACT FROM AUTHOR]

A total of 325 Listeria monocytogenes isolates from human, food and environmental sources were ribotyped with the Qualicon Automated Riboprinter Microbial Characterization System, using the enzyme EcoR1. The human isolates (n=137) represented all isolates from clinical cases in Norway from 1992 to 2005. The food and environmental isolates (n=188) were collected from different food related sources in Norway in the period 1989-2002. A total of 37 ribotypes were differentiated. Most common ribotypes (i.e., ribotypes represented by >5 isolates) were isolated from human as well as food and environmental sources. The exceptions were ribotypes DUP-1062D and 1058A, which were found only among food and environmental samples (14 and 5 times, respectively), and DUP-1042A, which was identified only among human clinical isolates. DUP-1030A, the most frequent ribotype among the human isolates, as well as ribotypes DUP-1042B, DUP-1042C, DUP-1049B and DUP-1062B were identified from both foods, environmental and human sources. DUP-1039C (n=54) and DUP-1045B (n=27) were frequently isolated from patients, food and environmental samples. The isolates were classified into lineages based on ribotyping results. The lineage I strain DUP-1038B was isolated every year from 1992 to 2005 from the human clinical samples. Out of 137 listeriosis cases, 76 (55.6%) were caused by lineage II strains. We found a considerable overlap between ribotypes, lineages and isolation sources. It does not seem possible to establish food strain specific regulations for L. monocytogenes based on ribotyping. [ABSTRACT FROM AUTHOR]

Campylobacteriosis is one of the most frequently occurring acute gastroenteritis in humans and 10% are caused by Campylobacter coli. A total of 136 isolates of C. coli from humans, poultry, and pigs were identified by multiplex polymerase chain reaction (PCR) and genetically characterized and compared by ribotyping. Automatic riboprints were performed with the PstI restriction enzyme and RiboPrinter®. All poultry, pig and human strains represented a heterogeneous spectre of ribotypes. Ten of 23 human strains (43%) could be given DUP-ID from the library represented by DUP-PSTI-1200 (n = 7), DUP-PST1-1201 (n = 2) and DUP-PSTI-1211 (n = 1). Eighteen of 28 (64%) poultry strains were given a DUP-ID. Three isolates were closely related to human strains DUP-PSTI-1201 (n = 2) and DUP-PSTI-1200 (n = 1) and may play an important role in the epidemiology of campylobacteriosis. Nineteen of 85 pig isolates (23%) could be given a DUP-ID, but none were common to human isolates. An overlap was found among poultry and pig isolates with DUP-PSTI-1182 and DUP-PSTI-1140. [ABSTRACT FROM AUTHOR]

Out of 125 samples we isolated 45 thermophilic Campylobacter spp. from Bø River by filtration followed by growth on selective agar plates. The species were identified by biotyping, multiplex PCR and riboprinting. C. jejuni represented 17 (37.7%), C. coli 2 (4.4%), C. lari 5 (11.1%), C. hyointestinalis 1 (2.2%) and C. species 20 (44.5%). Five of C. jejuni were classified in ribogroups (DUP-PST1-ID) while four C. lari were identified in three. The RiboPrinter® identified C. hyointestinalis. Dendrogram analysis grouped the isolates into two main clades and seven subclades. C. lari and C. coli were grouped in one clade while most C. spp. and the C. hyointestinalis were grouped in the other. C. jejuni were clustered in five subclades and display a high degree of diversity. Because of the frequent isolation of campylobacters in surface water and the lack of knowledge of the pathogenesis of Campylobacter strains and their virulence factors, special hygienic precautions should be taken to avoid the risk of transmitting campylobacteriosis from water. [ABSTRACT FROM AUTHOR]

Out of 100 faecal swab samples, 88 thermophilic Campylobacter representing ten herds were isolated. The species were identified by biotyping and multiplex polymerase chain reaction (PCR). Of the isolates, 86 (97.7%) were C. coli, one was C. lari, and one was C. jejuni. All three species were isolated from the same herd. Automatic riboprints were performed with the PstI restriction enzyme and RiboPrinter® (automated microbial characterization system). The identification of the isolates was predicted when the corresponding pattern matched one of the patterns in the DuPont identification (DUP-ID) library and was then assigned an identification number. A total of 81 selected C. coli isolates were riboprinted. The isolates could be divided into two clades and five subclades, and 19 isolates could be given five different DUP-IDs from the library: DUP-PST1-1182 (n=6), DUP-PST1-1208 (n=10), DUP-PST1-1175 (n=1), DUP-PST1-1140 (n=1), and DUP-PST1-1163 (n=1). Pigs are healthy carriers of C. coli which showed a high degree of diversity among and within the herds. [ABSTRACT FROM AUTHOR]

The purpose of this study was to use automated ribotyping procedure to track Listeria monocytogenes transmission in the cold smoked fish production chain and to characterize L. monocytogenes subtypes associated with the salmon processing industry. A total of 104 isolates, which had previously been obtained from a raw fish slaughter and processing plant (plant B) and an adjacent, downstream, salmon smoking operation (plant A), were characterized. These isolates had been obtained through a longitudinal study on Listeria presence, which covered a 31-week period, in both plants. Isolates had been obtained from samples taken from different machinery used throughout the production process. In addition, six isolates obtained from products produced in plant A two years after the initial study were included, so that a total of 110 isolates were characterized. Automated ribotyping was performed using both the restriction enzymes EcoRI and PvuII to increase the discriminatory power. The 110 L. monocytogenes isolates could be divided into 11 EcoRI ribotypes; PvuII ribotype data yielded multiple subtypes within 7 EcoRI ribotypes for a total of 21 subtypes based on both EcoRI and PvuII ribotyping. A total of three EcoRI ribotypes (DUP-1023C, DUP-1045B, and DUP-1053E) were isolated at multiple sampling times from both plants. In addition, one subtype (DUP-1053B) was isolated at multiple sampling times in only plant A, the salmon smoking operation. These data not only support that L. monocytogenes can persist throughout the salmon production system, but also showed that L. monocytogenes may be transmitted between slaughter and smoking operations or may be unique to smoking operations. While the majority of subtypes isolated have been rarely or never linked to human listeriosis cases, some subtypes have previously caused human listeriosis outbreaks and cases. Molecular subtyping thus is critical to identify L. monocytogenes transmission and niches in order to allow design and implementation of control strategies at the appropriate stage of production and in order to reduce the prevalence of L. monocytogenes linked to human disease. [ABSTRACT FROM AUTHOR]

Two plants processing salmon fillets and cold smoked salmon were investigated for occurrence of Listeria in products and the environment. Analyses were conducted for a period of 31 weeks. At plant A, 252 samples were examined of which 97 were from unprocessed fish and 155 from cold-smoked fish. At plant B, 189 samples of unprocessed fish were investigated. The first examination of unprocessed fish at plant A showed a presence of L. monocytogenes and L. spp. in 81% and 19% of the samples respectively. For cold-smoked fish the figures were 43% and 23%. At plant B, L. monocytogenes was isolated in 63% of the samples. During the test period, management at the processing plant initiated various hygiene precautions to improve the sanitary situation. The last batch of analyses of unprocessed fish at plant A showed a presence of L. monocytogenes and L. spp. in 42% and. 33% of the samples respectively. For cold-smoked fish, the figures were 6% and 11%. The isolation figures at plant B for L. monocytogenes and L. spp. were 50% and 17% respectively. The hygienic precautions did not have a significant effect on the presence of L. monocytogenes and L. spp. We suggest that Listeria bacteria are a part of the resident flora and are not eliminated by current cleaning and sanitation programmes. Cold-smoking, however, gave a significant reduction in the isolation of L. monocytogenes (P?=?0.0082), while the isolation of L. spp. did not decrease after this process. [ABSTRACT FROM AUTHOR]

Campylobacteriosis caused by Campylobacter jejuni, Campylobacter coli and Campylobacter lari is one of the most frequently occurring acute gastroenteritis diseases in humans. An important risk factor is untreated drinking water. Thermotolerant campylobacters were isolated from Bø River water using 100-ml filtrate samples. Campylobacters were isolated from 32 of 60 samples (53.3%). Of the 75 strains isolated, 47 (62.7%) were detected using the enrichment procedure and 28 (37.2%) by direct plating of the filter. Campylobacter coli was the dominating species (44%) followed by Campylobacter jejuni (34.6%) and Campylobacter lari (14.7%). Five isolates (6.7%) could not be classified. Campylobacter were isolated without the occurrence of fecal coliforms indicating that fecal coliforms cannot be used as index bacteria to predict the occurrence of campylobacters. Case-control studies have indicated untreated drinking water and barbecuing as important risk factors, and waterborne campylobacteriosis has been caused by the three species isolated. The virulence factors of isolated environmental strains are not fully known. It was concluded that it is hazardous to use untreated surface water for drinking. Special efforts should be taken to avoid contamination of agricultural products from irrigation. [ABSTRACT FROM AUTHOR]

Expansion of the range of Ixodes ricinus ticks further north and to higher altitudes affects the spread of tick-borne encephalitis virus (TBEV) in new territories and increases the risk of human infections. Over the past decade, the number of tick-borne encephalitis (TBE) cases has increased in Norway and parts of the southern coast have been established as endemic. The aims of this study were to confirm the existence of TBEV endemic foci, determine the spread of TBE in different localities of Norway, review and compare relevant published records on the prevalence of TBEV in the different developmental stages and sex of the ticks. Ticks were collected from nine locations along the southern coast of Norway during June-July 2009. For the detection of TBEV-specific RNA, a real-time RT-PCR targeting a part of the 3’ non-coding region of the TBEV genome was used. A total of 1542 ticks grouped in 250 pools were examined and TBEV RNA was detected in 5 pools with overall prevalence of 0.32%. [ABSTRACT FROM AUTHOR]

Abstract: Lyme borreliosis (LB) caused by the spirochete Borrelia burgdorferi sensu lato is the most common tick-borne zoonosis in the Northern Hemisphere. B. burgdorferi s.l. can infect humans and wild and domestic animals. Ixodes ricinus is the main vector, and small rodents are the most important mammalian reservoirs hosts of B. burgdorferi s.l. in Europe. The prevalence of B. burgdorferi s.l. in I. ricinus ticks from captured rodents, calculated specific infectivities, and transmission coefficients were estimated in order to investigate the role of voles and mice in transmission of the LB causative agent. A total of 12.3% (53 out of 431) of immature I. ricinus ticks from rodents in Lithuania and 3.25% (21 out of 646) in Norway were infected with B. burgdorferi s.l. In Lithuania a total of 40% infested Microtus arvalis, 29% of Myodes glareolus and 4.8% of Apodemus flavicollis carried infected larvae and 67% of M. glareolus, 36% of M. arvalis but none of A. flavicollis carried infected nymphs. In Norway, 2.4% of larvae and 12.1% of nymphs feeding on A. flavicollis were infected. A total of 9% of infested A. flavicollis carried infected larvae and 13% – infected nymphs. Borrelia afzelii was the single genospecies identified in ticks feeding on rodents in Lithuania, and was predominant in ticks collected from rodents in Norway. According to calculated indices of specific infectivity and tick-to host transmission coefficient, M. arvalis and M. glareolus voles were found to be more efficient in transmitting B. burgdorferi s.l. to ticks than A. flavicollis mice. GLMM analysis showed that rodent species significantly influenced the probability of a larva being infected with B. burgdorferi s.l. The larvae feeding on M. arvalis and M. glareolus were more likely to be infected with B. burgdorferi s.l. than those feeding on A. flavicollis. This is the first study to report the quantitative roles of voles and mice in the transmission of B. burgdorferi s.l. to larval ticks in Lithuania and Norway. [Copyright &y& Elsevier]

Abstract: The zoonotic rickettsial pathogen Anaplasma phagocytophilum has a broad geographic distribution and a high degree of biological and clinical diversity. To investigate the genetic diversity of A. phagocytophilum strains in the Baltic region and Norway, three species of Ixodidae ticks were examined for A. phagocytophilum infection, and two genes of the pathogen genome were analyzed. Analysis of partial 16S rRNA and partial major surface protein (msp4) gene sequences was accomplished through nested PCR and sequence analysis. Strains identified in this study were compared with those originating from other European countries and the United States. Seven 16S rRNA gene variants and fifteen msp4 gene variants of A. phagocytophilum were detected. Nine sequences had unique nucleotide polymorphisms and therefore differed from other A. phagocytophilum sequences previously submitted to GenBank. The present study represents the first molecular characterization of A. phagocytophum strains circulating in Lithuania and describes the strains detected in Ixodes persulcatus ticks in Latvia and Estonia. This is also the first report describing A. phagocytophilum strains isolated from Dermacentor reticulatus ticks. [Copyright &y& Elsevier]

Lyme borreliosis (LB) caused by the spirochete Borrelia burgdorferi s.l. is the most frequently diagnosed tick-bome zoonozis in Europe, North America, and Asia. B. burgdorferi s.l. can infect humans and wild and domestic animals. Small rodents are the most important reservoir host of B. burgdorferi s.l. The aim of present study was to detect f B. burgdorferi s.l. in different tissue samples of small rodents and identify Borrelia genospecies and strains using different molecular markers and detection methods. Flagellin (fla) and outer surface protein A (ospA) encoding genes of Borrelia genome were used as targets for PCR amplification. We found that the ospA gene was a more sensitive marker for the detection of B. burgdorferi s.l. than fla gene. The presence of B. burgdorferi s.l. in urinary bladder, spleen and ear biopsy samples of 136 small rodents was compared. B. burgdorferi s.l. infection was detected with different rates in ear and bladder tissues, but was not found in spleen samples. Multiplex PCR assay based on ospA gene was used for identifying B. burgdorferi s. s., B. afzelii, and B. garinii genospecies. B. afzelii was a single genospecies detected in small rodents from Norway and was predominant in rodents from Lithuania. B. garinii strains from rodents were identified by nucleotide sequencing of PCR products. Phylogenetic relationship between B. garinii strains and their correspondence to OspA serotype types were compared with the sequences registered in GenBank database. B. garinii strains detected in the present study showed similarities with sequences of OspA serotype 5 type and OspA serotype 6 type. [ABSTRACT FROM AUTHOR]

During the last centuries, red deer (Cervus elaphus) have survived a drastic decline in population number which has gone as far as extinction in many regions of the continent. Today's deer population is abundant and widespread, however, affected by re-introduction and keeping isolated animals in enclosures. It is necessary to observe the genetics of these animals and to monitor changes that occur in populations as well as apply the obtained knowledge in creating and restoring the species. This study investigates the levels of genetic diversity of red deer. Muscle and liver samples were obtained from 27 individuals corresponding to 2 red deer populations from Lithuania and Norway. Samples were analyzed by RAPD (random amplified polymorphic DNA) polymerase chain reaction (PCR) using ten primers (ROTH-180-01, ROTH-180-02, ROTH-180-03, ROTH-180-04, ROTH-180-05, ROTH-180-06, ROTH-180-07, ROTH-180-08, ROTH-180-09, ROTH-180-10). PCR products were sorted according to their size by electrophoresis in 1.7% agarose gel. Gel electrophoresis showed that there were 64 polymorphic loci with the fragment size varying from 150 to 3 000. The "TREECON for Windows" programme was used for the estimation of genetic variability. Genetic variability among the red deer separate individuals varied from 0.08 to 0.80. [ABSTRACT FROM AUTHOR]

A total of 693 small rodents were live-trapped in Lithuania and Norway. In Lithuania, the mice Apodemus flavicollis and A. agrarius were more frequently infested with ticks than the voles Microtus arvalis and Myodes glareolus. The overall prevalence of infestation with Ixodes ricinus in different sampling locations of Lithuania ranged for A. flavicollis from 0% to 92.3%, for M. glareolus from 5.9% to 100%, and for M. arvalis from 0% to 37.8%. In Norwegian sites the prevalence of infestation was higher for A. flavicollis (62.7.98% in different sites) than for A. sylvaticus (42.9.77.8%). The most intensive infestation of rodents with I. ricinus larvae was observed in May and June. Contrasting patterns of infestation with larvae and nymphs were observed in males and females of M. glareolus and A. flavicollis. [ABSTRACT FROM AUTHOR]

Background: Ixodes ricinus ticks transmit Babesia species to vertebrate hosts. Using molecular tools we were able to detect the presence of this piroplasmid in its vector. The aims of this study were to investigate the prevalence and identity of Babesia species in questing ticks collected in various areas of Norway. Methods: DNA from questing l. ricinus ticks were examined with a realtime PCR for the presence of Babesia. Positive samples of tick DNA were identified to species using PCR, and sequence analysis. Results: From a total of 1908 questing l. ricinus ticks, 17 (0.9%) indicated the presence of Babesia spp. after realtime- PCR screening. Ixodes ricinus harbouring Babesia spp. was detected in 9 out of 22 localities. Further molecular analyses of DNA from these positive ticks indicate the presence of Babesia venatorum, B. divergens, B. capreoli and a currently undescribed Babesia in Norwegian ticks. The most prevalent was B. venatorum found in 71% of the positive ticks. Conclusions: A total of 17 out of 1908 (0.9%) ticks were positive for Babesia. Our data confirm that there are several Babesia species in ticks in Norway. Babesia venatorum was the most prevalent. This species has a zoonotic potential and may cause human babesiosis following a tick bite [ABSTRACT FROM AUTHOR]

Five salmon processing plants were investigated for periods of eight (plants A and B) and four (plants C, D and E) months for the occurrence of Listeria monocytogenes. A total of 226 strains of L. monocytogenes were isolated and subtyped. Automated ribotyping with DuPont Qualicon RiboPrinter® system with the enzyme EcoR1 was used to differentiate and characterize isolates for simple molecular tracking. The isolated strains could be divided into 16 DuPont ribotypes. The ribotypes DUP-1023C, DUP-1039C, DUP-1044E, DUP-1045B, DUP-1046A, DUP-1062B and 1062C were found to be persistent in the plants, as they were found through the whole sampling period. The remaining 9 ribotypes were isolated sporadically. Five of the persistent subtypes (DUP-1023C, DUP-1039C, DUP-1046A, DUP-1062B and 1062C) where isolated in high numbers. Ribotype DUP-1045B and DUP-1039C were found in four of the five investigated plants. [ABSTRACT FROM AUTHOR]

The use of molecular methods such as species-specific PCR, Multiplex-PCR, RT-PCR, reverse line blot hybridisation in investigations of tick-borne pathogens allowed to detect and identify the causative agents of Lyme borreliosis, anaplasmosis, and babesiosis in ticks and rodents in Lithuania and Norway. The overall prevalence of Borrelia burgdorferi s. l. infection detected in Lithuanian and Norwegian ticks was found to be 13.3% (223/1679) and 5.4% (75 /1383), respectively. A total 68 of 398 (17.1%) rodent ear extractions screened by PCR were found positive for B. burgdorferi s. l. infection. B. burgdorferi s. l. was detected in 53 of 428 (12.4%) immature Ixodes ricinus ticks collected on rodents in Lithuania and in 30 of 782 (3.8%) collected on rodents in Norway. In 24 of 173 (13.8%) ticks feeding on passerine migrating birds collected in Norway, B. burgdorferi s. l. pathogens were detected. Three clinically important species (B. afzelii, B. garinii and B. burgdorferi s. s.) were identified in ticks and rodents. Anaplasma sp. was detected in 5% of questing ticks and 19.6% of ticks collected from birds in Lithuania. A. phagocytophilum pathogens were detected in 7.1% of ticks from birds and in 93 of 1634 (5.7%) I. ricinus ticks collected from vegetation in Norway. To Babesia divergens, positive were 2% and 0.9% of questing ticks collected in Lithuania and Norway respectively. [ABSTRACT FROM AUTHOR]