Comparison of virulence of Francisella tularensis ssp. holarctica genotypes B.12 and B.FTNF002-00.

BACKGROUND: Two main genetic groups (B.12 and B.FTNF002-00) of Francisella tularensis ssp. holarctica are endemic in Europe. The B.FTNF002-00 group proved to be dominant in Western European countries, while strains of the B.12 group were isolated mainly in Northern, Central and Eastern Europe. The clinical course of tularemia in the European brown hare (Lepus europaeus) also shows distinct patterns according to the geographical area. Acute course of the disease is observed in hares in Western European countries, while signs of sub-acute or chronic infection are more frequently detected in the eastern part of the continent. The aim of the present study was to examine whether there is any difference in the virulence of the strains belonging to the B.FTNF002-00 and B.12 genetic clades. RESULTS: Experimental infection of Fischer 344 rats was performed by intra-peritoneal injection of three dilutions of a Hungarian (B.12 genotype) and an Italian (B.FTNF002-00 genotype) F. tularensis ssp. holarctica strain. Moderate difference was observed in the virulence of the two genotypes. Significant differences were observed in total weight loss values and scores of clinical signs between the two genotypes with more rats succumbing to tularemia in groups infected with the B.FTNF002-00 genotype. CONCLUSIONS: Results of the experimental infection are consistent with previous clinical observations and pathological studies suggesting that F. tularensis ssp. holarctica genotype B.FTNF002-00 has higher pathogenic potential than the B.12 genotype.

First detection of Pasteurella multocida type B:2 in Hungary associated with systemic pasteurellosis in backyard pigs.

This is the first report of Pasteurella multocida type B in Hungarian pigs. This disease was observed in backyard-raised pigs in three households within a small area. Neither the source of the infection nor the epidemiological connection between any of the premises could be determined. The most consistent lesion was dark red discolouration of the skin of the ventral neck and brisket, with accompanying oedema and haemorrhages. The morbidity was low and lethality relatively high, with three dead (50%) and two euthanised (33%) out of six affected animals. A total of three isolates of P. multocida (P55, P56 and P57) were cultured from these cases and examined in detail. These were identified as P. multocida ssp. multocida biovar 3. All were toxA negative and belonged to serotype B:2. Multilocus sequence typing was used to assign these to a new sequence type (ST61) that is closely related to other haemorrhagic septicaemia causing strains of P. multocida regardless of the host. M13 polymerase chain reaction and virulence-associated gene typing also show that type B strains form a highly homogeneous, distinct phylogenic group within P. multocida.

Worldwide phylogenetic relationship of avian poxviruses.

Poxvirus infections have been found in 230 species of wild and domestic birds worldwide in both terrestrial and marine environments. This ubiquity raises the question of how infection has been transmitted and globally dispersed. We present a comprehensive global phylogeny of 111 novel poxvirus isolates in addition to all available sequences from GenBank. Phylogenetic analysis of the Avipoxvirus genus has traditionally relied on one gene region (4b core protein). In this study we expanded the analyses to include a second locus (DNA polymerase gene), allowing for a more robust phylogenetic framework, finer genetic resolution within specific groups, and the detection of potential recombination. Our phylogenetic results reveal several major features of avipoxvirus evolution and ecology and propose an updated avipoxvirus taxonomy, including three novel subclades. The characterization of poxviruses from 57 species of birds in this study extends the current knowledge of their host range and provides the first evidence of the phylogenetic effect of genetic recombination of avipoxviruses. The repeated occurrence of avian family or order-specific grouping within certain clades (e.g., starling poxvirus, falcon poxvirus, raptor poxvirus, etc.) indicates a marked role of host adaptation, while the sharing of poxvirus species within prey-predator systems emphasizes the capacity for cross-species infection and limited host adaptation. Our study provides a broad and comprehensive phylogenetic analysis of the Avipoxvirus genus, an ecologically and environmentally important viral group, to formulate a genome sequencing strategy that will clarify avipoxvirus taxonomy.

Antibiotic susceptibility profiles of Mycoplasma bovis strains isolated from cattle in Hungary, Central Europe.

BACKGROUND: Mycoplasma bovis is a worldwide pathogen, causative agent of pneumonia, mastitis, arthritis, and a variety of other symptoms in cattle. The economic losses due to mycoplasma pneumonia could be reduced by antibiotic treatment. The aim of the present study was to determine the in vitro susceptibility of M. bovis strains isolated from cattle in Hungary to eleven antibiotics. RESULTS: Minimal inhibitory concentration (MIC) values of 35 M. bovis strains collected from different parts of Hungary between 2010 and 2013 were determined by the microbroth dilution method. Strains with high MIC values were found in the case of all applied antibiotics. The most effective antibiotics tested in vitro were fluoroquinolones (MIC90 danofloxacin 0.312 µg/ml, enrofloxacin 0.312 µg/ml, marbofloxacin 0.625 µg/ml). Our results confirm the observations of increasing MIC values to antibiotics commonly used in the therapy of mycoplasma infections, primarily to tetracyclines; tetracycline (MIC90 16 µg/ml) and oxytetracycline (MIC90 ≥ 64 µg/ml) and macrolides; tylosin (MIC90 ≥ 128 µg/ml) and tilmicosin (MIC90 ≥ 128 µg/ml). The growth of many M. bovis strains was not inhibited by gentamicin (MIC90 8 µg/ml), spectinomycin (MIC90 ≥ 256 µg/ml), florfenicol (MIC90 8 µg/ml) or lincomycin (MIC90 ≥ 64 µg/ml). CONCLUSIONS: Our results emphasize the necessity of periodic testing for antibiotic susceptibility in this geographic region. Based on our in vitro examinations, fluoroquinolones could be the most effective drugs for the therapy of M. bovis infections in Hungary. However, current antimicrobial use policies have to be taken into account to avoid further antibiotic resistance development and to reserve fluoroquinolones for the treatment of severe infections which have responded poorly to other classes of antimicrobials.

Phylogeny of Mycoplasma bovis isolates from Hungary based on multi locus sequence typing and multiple-locus variable-number tandem repeat analysis.

BACKGROUND: Mycoplasma bovis is an important pathogen causing pneumonia, mastitis and arthritis in cattle worldwide. As this agent is primarily transmitted by direct contact and spread through animal movements, efficient genotyping systems are essential for the monitoring of the disease and for epidemiological investigations. The aim of this study was to compare and evaluate the multi locus sequence typing (MLST) and the multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) through the genetic characterization of M. bovis isolates from Hungary. RESULTS: Thirty one Hungarian M. bovis isolates grouped into two clades by MLST. Two strains had the same sequence type (ST) as reference strain PG45, while the other twenty nine Hungarian isolates formed a novel clade comprising five subclades. Isolates originating from the same herds had the same STs except for one case. The same isolates formed two main clades and several subclades and branches by MLVA. One clade contained the reference strain PG45 and three isolates, while the other main clade comprised the rest of the strains. Within-herd strain divergence was also detected by MLVA. Little congruence was found between the results of the two typing systems. CONCLUSIONS: MLST is generally considered an intermediate scale typing method and it was found to be discriminatory among the Hungarian M. bovis isolates. MLVA proved to be an appropriate fine scale typing tool for M. bovis as this method was able to distinguish closely related strains isolated from the same farm. We recommend the combined use of the two methods for the genotyping of M. bovis isolates. Strains have to be characterized first by MLST followed by the fine scale typing of identical STs with MLVA.

Antimicrobial susceptibility of Francisella tularensis subsp. holarctica strains from Hungary, Central Europe.

OBJECTIVES: Determining the in vitro susceptibility to 11 antibiotics of Francisella tularensis subsp. holarctica strains belonging to the phylogenetic group B.13, from different areas of Hungary. METHODS: Twenty-nine F. tularensis strains isolated between 2003 and 2010 from free-ranging European brown hares (Lepus europaeus) and a captive patas monkey (Erythrocebus patas) were collected from different parts of Hungary and examined for antibiotic susceptibility with commercially available MIC test strips on modified Francis agar plates; values were interpreted according to CLSI breakpoints. RESULTS: The strains were susceptible to aminoglycosides (MIC(90) values: gentamicin, 0.75 mg/L; and streptomycin, 6.0 mg/L), tetracyclines (MIC(90) values: tetracycline, 0.5 mg/L; and doxycycline, 1.0 mg/L), quinolones (MIC(90) values: ciprofloxacin, 0.047 mg/L; and levofloxacin, 0.023 mg/L) and chloramphenicol (MIC(90) value: 1.5 mg/L), i.e. antibiotics commonly used in therapy. Tigecycline (MIC(90) value: 0.19 mg/L) and rifampicin (MIC(90) value: 1.0 mg/L) were also active against F. tularensis strains, while resistance to erythromycin (MIC(90) value: >256 mg/L) and linezolid (MIC(90) value: 32 mg/L) was observed in all strains. CONCLUSIONS: Based on the results, quinolones are recommended as first choice therapy for F. tularensis infection. The in vitro susceptibility of the strains to tigecycline may encourage the application of this antibiotic as well. The similar antibiotic susceptibilities of the Hungarian strains belonging to different subclades of phylogenetic group B.13 indicates that strains from other Central and Eastern European countries belonging to this group might also have the same susceptibility profile.

First description of hypospadias and inguinal bilateral cryptorchidism in mouflon (Ovis Gmelini Musimon) - short communication.

A mouflon showing severe weight loss and cachexia was examined. The animal had horns and a male-like body frame but the preputial fur was missing. The scrotum was completely absent. Both testicles were located next to the inguinal canal under the skin, and appeared only slightly smaller than normal. The prepuce was located in a perianal position under the anus and it resembled a vulva. The penis was underdeveloped and curled up inside the prepuce like an enlarged clitoris. The bulbourethral region and the urinary bladder looked normal. The orifice of the urethra was located in its normal position, but the glans penis appeared deformed. Microscopic examination of the testicles revealed mild degeneration of the seminiferous tubules and a marked proliferation of the interstitial connective tissue with Leydig cells still present. There were no marked changes in the bulbar part of the penis but the apical part exhibited marked interstitial fibrosis. This is the first description of hypospadias and cryptorchidism in mouflon.

Toxoplasmosis in Tammar wallabies (Macropus eugenii) in the Budapest Zoo and Botanical Garden (2006-2010).

Smaller macropodid species (commonly referred to as wallabies) are extremely susceptible to toxoplasmosis: in most cases, infection with Toxoplasma gondii leads to death within a short time. Between June 2006 and July 2010, T. gondii was detected by immunohistochemical examination in six Tammar wallabies (Macropus eugenii) that died in the Budapest Zoo and Botanical Garden; in another four specimens histopathology revealed T. gondii-like organisms (which could not be differentiated from Neospora caninum solely by morphology), and in another 11 animals toxoplasmosis as the possible cause of death could not be excluded. The current zoo population of 12 Tammar wallabies was tested for T. gondii IgG antibodies by the modified agglutination test (MAT), with negative results. We suppose that most of the deaths were due to acute toxoplasmosis resulting from a recent infection.

Genomic evolution of avian polyomaviruses with a focus on budgerigar fledgling disease virus.

Gammapolyomaviruses may cause serious inflammatory diseases in a broad range of avian hosts. In this study we investigated genomic evolution of and selection constraint acting on avian polyomaviruses (APyVs). Our analyses suggested that goose haemorrhagic polyomavirus (GHPV) evolves more slowly (3.03 × 10-5 s/s/y mean evolutionary rate) than budgerigar fledgling disease virus (BFDV), finch polyomavirus (FPyV) and canary polyomavirus (CaPyV) (1.39 × 10-4 s/s/y, 2.63 × 10-4 s/s/y and 1.41 × 10-4 s/s/y mean evolutionary rate, respectively). In general, purifying selection seems to act on the protein coding regions of APyV genomes, although positive Darwinian selection was also predicted in a few positions (e.g., in the large tumor antigen coding region of BFDV and GHPV and in the capsid protein sequences of BFDV). The importance of these aa changes remains elusive. Overall, a better understanding of adaptive changes in the genome of APyVs requires additional data from various incidental hosts and reservoir species.

Tissue tropism of highly pathogenic avian influenza virus subtype H5N1 in naturally infected mute swans (Cygnus Olor ), domestic geese (Aser Anser var. domestica), pekin ducks (Anas platyrhynchos) and mulard ducks ( Cairina moschata x anas platyrhynchos).

The 2006 epidemic due to highly pathogenic avian influenza virus (HPAIV) subtype H5N1 in Hungary caused the most severe losses in waterfowl which were, according to the literature at the time, supposed to be the most resistant to this pathogen. The presence of pathological lesions and the amount of viral antigen were quantified by gross pathology, histopathology and immunohistochemistry (IHC) in the organs of four waterfowl species [mute swans (n = 10), domestic geese (n = 6), mulard ducks (n = 6) and Pekin ducks (n = 5)] collected during the epidemic. H5N1 subtype HPAIV was isolated from all birds examined. Quantitative real-time reverse transcriptase-polymerase chain reaction (qRRT-PCR) was also applied on a subset of samples [domestic geese (n = 3), mulard (n = 4) and Pekin duck (n = 4)] in order to compare its sensitivity with IHC. Viral antigen was detected by IHC in all cases. However, the overall presence of viral antigen in tissue samples was quite variable: virus antigen was present in 56/81 (69%) swan, 22/38 (58%) goose, 28/46 (61%) mulard duck and 5/43 (12%) Pekin duck tissue samples. HPAIV subtype H5N1 was detected by qRRT-PCR in all birds examined, in 19/19 (100%) goose, 7/28 (25%) mulard duck and 12/28 (43%) Pekin duck tissue samples. As compared to qRRTPCR, the IHC was less sensitive in geese and Pekin ducks but more sensitive in mulard ducks. The IHC was consistently positive above 4.31 log10 copies/reaction but it gave very variable results below that level. Neurotropism of the isolated virus strains was demonstrated by finding the largest amount of viral antigen and the highest average RNA load in the brain in all four waterfowl species examined.

Mandibular Ossifying Fibroma and Multiple Oral Papillomas in a Roe Deer (Capreolus capreolus).

An emaciated, adult, free-ranging roe deer (Capreolus capreolus) presenting a large mandibular mass, was shot by a game warden in Sissach, Switzerland. The head of the roe deer was submitted to the Center for Fish and Wildlife Health for examination. Grossly, the mass consisted of a 6 × 7 × 4 cm mandibular exophytic growth, associated with loss of incisors teeth. On cut section, a hard, light-tan core was rimmed by a thick layer of soft tissue. Computed tomography examination confirmed the mandibular origin of the mass. Histologically, the mass consisted of an unencapsulated fibro-osseous neoplasm. The bony portion was composed of multiple haphazardly arranged spicules rimmed by osteoblasts with no associated periosteal layer. Embedding the bony spicules were short anastomosing and branching streams and bundles of spindled cells. The overlaying partially ulcerated mucosa, showed prominent rete ridges deepening into the submucosa. In addition to the mandibular mass, multiple soft cauliflower-like proliferations were expanding from the gingival surface. Histologically, these masses were arranged in papillary elements composed of pluristratified squamous epithelium with long rete ridges extending into a rich underlying fibrovascular supportive stroma. Neither papillomaviral DNA nor antigen could be identified in association with the oral masses. The gross, histological and radiological features of the mandibular mass are consistent with an ossifying fibroma, while the cauliflower oral masses were diagnosed as papillomas.

Different dynamics of Usutu virus infections in Austria and Hungary, 2017-2018.

Usutu virus (USUV), a mosquito-borne flavivirus closely related to West Nile virus, emerged in Austria in 2001, when it caused a considerable mass-mortality of Eurasian blackbirds. Cases in birds increased until 2003 and quickly declined thereafter, presumably due to developing herd immunity. Since 2006, no further cases were recorded, until two blackbirds were tested positive in 2016. In Hungary, USUV first appeared in 2005 and has caused only sporadic infections since then. Initially, the only genetic USUV lineage found across both countries was Europe 1. This changed in 2015/2016, when Europe 2 emerged, which has since then become the prevalent lineage. Due to dispersal of these strains and introduction of new genetic lineages, USUV infections are now widespread across Europe. In 2009, the first cases of USUV-related encephalitis were described in humans, and the virus has been frequently detected in blood donations since 2016. To monitor USUV infections among the Austrian wild bird population in 2017/2018, 86 samples were investigated by RT-PCR. In 67 of them, USUV nucleic acid was detected (17 in 2017, 50 in 2018). The majority of succumbed birds were blackbirds, found in Vienna and Lower Austria. However, the virus also spread westwards to Upper Austria and southwards to Styria and Carinthia. In Hungary, 253 wild birds were examined, but only six of them were infected with USUV (five in 2017, one in 2018). Thus, in contrast to the considerable increase in USUV-associated bird mortality in Austria, the number of infections in Hungary declined after a peak in 2016. Except for one case of USUV lineage Africa 3 in Austria in 2017, Europe 2 remains the most prevalent genetic lineage in both countries. Since USUV transmission largely depends on temperature, which affects vector populations, climate change may cause more frequent USUV outbreaks in the future.

Detection of African swine fever virus in cell culture and wild boar tissues using a commercially available monoclonal antibody.

Following the introduction of African swine fever virus (ASFV) into Europe in 2007, ASFV infection has spread continuously over the past years and it became a high level disease threat in Europe and also Asia. Examination of suspect clinical cases for ASF with rapid and sensitive laboratory methods can substantially contribute to the detection and characterization of new outbreaks. In this study two sensitive tests were developed for the detection of the p72 major capsid protein of ASFV both in cell culture with an immunocytochemical (IC) and in tissue samples with an immunohistochemical (IHC) method using a commercially available mouse monoclonal antibody (clone 1BC11). The IC test was able to detect the virus at high virus dilutions in cell culture and the IHC test indicated the presence of ASFV in all formalin-fixed and paraffin-embedded tissue samples collected from two wild boars. The reported IC and IHC methods were found to be useful ancillary laboratory tests for research purposes and for the diagnosis of acute ASF.

Multi-Approach Investigation Regarding the West Nile Virus Situation in Hungary, 2018.

The West Nile virus is endemic in multiple European countries and responsible for several epidemics throughout the European region. Its evolution into local or even widespread epidemics is driven by multiple factors from genetic diversification of the virus to environmental conditions. The year of 2018 was characterized by an extraordinary increase in human and animal cases in the Central-Eastern European region, including Hungary. In a collaborative effort, we summarized and analyzed the genetic and serologic data of WNV infections from multiple Hungarian public health institutions, universities, and private organizations. We compared human and veterinary serologic data, along with NS5 and NS3 gene sequence data through 2018. Wild birds were excellent indicator species for WNV circulation in each year. Our efforts resulted in documenting the presence of multiple phylogenetic subclades with Balkans and Western-European progenitor sequences of WNV circulating among human and animal populations in Hungary prior to and during the 2018 epidemic. Supported by our sequence and phylogenetic data, the epidemic of 2018 was not caused by recently introduced WNV strains. Unfortunately, Hungary has no country-wide integrated surveillance system which would enable the analysis of related conditions and provide a comprehensive epidemiological picture. The One Health approach, involving multiple institutions and experts, should be implemented in order to fully understand ecological background factors driving the evolution of future epidemics.

Avian bornaviruses in psittacine birds from Europe and Australia with proventricular dilatation disease.

To determine whether avian bornaviruses (ABVs) were a factor in proventricular dilatation disease (PDD), we used immunohistochemistry, reverse transcription-PCR, and nucleotide sequence analysis to examine paraffin wax-embedded or frozen tissue samples of 31 psittacine birds with this disease. PDD is a fatal disease of psittacine birds associated with nonsuppurative encephalitis and ganglioneuritis of the upper intestinal tract. Tissue samples had been collected from 1999 through 2008 in Austria, Switzerland, Hungary, and Australia. Immunohistochemical demonstration of viral antigen within the brain and vegetative nerve system of the gastrointestinal tract provides strong evidence for a causative role of ABVs in this condition. Partial sequences of nucleoprotein (p40) and matrix protein (gp18) genes showed that virus in most of our cases belonged to the ABV-2 and ABV-4 groups among the 5 genogroups described so far. Viral sequences of 2 birds did not match any of the described sequences and clustered together in a new branch termed ABV-6.

Molecular analysis of the VP7 gene of pheasant rotaviruses identifies a new genotype, designated G23.

Rotavirus-associated enteritis has been reported in pheasants, but there is no information on the genetic/antigenic features of pheasant rotaviruses. In this study, we sequenced the VP7-encoding genome segment of three pheasant rotavirus strains detected during 2008 in Hungary. The full-length genome segment was 1,070 bp long, while the open reading frame was predicted to encode a 330-aa-long protein. The nucleotide sequence identities among the three pheasant rotavirus strains were high (> or =94%), whereas the range of nucleotide sequence identities to other avian and mammalian rotavirus VP7 genes fell between 68 and 73% and between 60 and 66%, respectively. Our findings indicate that these Hungarian pheasant rotaviruses need to be considered representatives of a new VP7 genotype specificity, designated G23.

Generalized tularemia in a vervet monkey (Chlorocebus aethiops) and a patas monkey (Erythrocebus patas) in a zoo.

Generalized tularemia was diagnosed in a vervet monkey (Chlorocebus aethiops) and a patas monkey (Erythrocebus patas), both of which died suddenly in the Szeged Zoo, Szeged, Hungary. Macroscopic lesions in each animal included disseminated, grayish-white foci in the lungs, lymph nodes, spleen, liver, and kidney. All focal lesions were characterized microscopically as purulent to pyogranulomatous to granulomatous inflammation with necrosis. Francisella tularensis subsp. holarctica strains were isolated from tissue samples on modified Francis agar after mouse passage and identified by a commercial carbon-source utilization test and polymerase chain reaction-based amplification and sequencing of a portion of the 16S ribosomal RNA gene.

Papillomavirus-associated fibropapillomas of red deer ( Cervus elaphus ).

Oval, firm, cutaneous tumours with a rough, hairless, pigmented surface, exhibiting a moderately pronounced papillary structure were detected on the abdominal skin of two young red deer ( Cervus elaphus ). One animal was shot in Lower Austria in 2004, the other at a deer farm in Hungary in 2007. Histological examination of both samples classified the tumours as fibropapillomas, showing marked proliferation of fibroblasts and connective tissue, accompanied by hyperkeratosis, parakeratosis and acanthosis of the overlaying epidermis, and occasional foci of inflammation. The distribution of cytokeratin and vimentin was characterised in the lesion. The presence of papillomavirus (PV) antigen was demonstrated by immunohistochemistry in both cases. Papillomavirus-specific DNA was successfully amplified by PCR from one sample. The obtained partial nucleotide sequence of the L2 ORF exhibited the highest critical identity values with the homologous regions of Delta-papillomaviruses, especially the Roe deer papillomavirus (93%). Phylogenetic analysis of the partial L2 ORF sequence alignment of 10 papillomaviruses by both neighbour-joining and maximum parsimony method confirmed that the Red deer PV is very closely related to the Western roe deer papillomavirus (CcPV1).

Frequent rearrangement may explain the structural heterogeneity in the 11th genome segment of lapine rotaviruses - short communication.

In rotaviruses, intragenic recombination or gene rearrangement occurs almost exclusively in the genome segments encoding for non-structural proteins. Rearranged RNA originates by mechanisms of partial sequence duplications and deletions or insertions of non-templated nucleotides. Of interest, epidemiological investigations have pointed out an unusual bias to rearrangements in genome segment 11, notably in rotavirus strains of lapine origin, as evidenced by the detection of numerous lapine strains with super-short genomic electropherotype. The sequence of the full-length genome segment 11 of two lapine strains with super-short electropherotype, LRV-4 and 3489/3, was determined and compared with rearranged and normal cognate genome segments of lapine rotaviruses. The rearranged genome segments contained head-to-tail partial duplications at the 3' end of the main ORF encoding NSP5. Unlike the strains Alabama and B4106, intermingled stretches of non-templated sequences were not present in the accessory RNA of LRV-4 and 3489/3, while multiple deletions were mapped, suggesting the lack of functional constraints. Altogether, these findings suggest that independent rearrangement events have given origin to the various lapine strains that have super-short genome pattern.

Combination therapy of rabies-infected mice with inhibitors of pro-inflammatory host response, antiviral compounds and human rabies immunoglobulin.

Recent studies demonstrated that inhibitors of pro-inflammatory molecular cascades triggered by rabies infection in the central nervous system (CNS) can enhance survival in mouse model and that certain antiviral compounds interfere with rabies virus replication in vitro. In this study different combinations of therapeutics were tested to evaluate their effect on survival in rabies-infected mice, as well as on viral load in the CNS. C57Bl/6 mice were infected with Silver-haired bat rabies virus (SHBRV)-18 at virus dose approaching LD50 and LD100. In one experimental group daily treatments were initiated 4 h before-, in other groups 48 or 96 h after challenge. In the first experiment therapeutic combination contained inhibitors of tumour necrosis factor-α (infliximab), caspase-1 (Ac-YVAD-cmk), and a multikinase inhibitor (sorafenib). In the treated groups there was a notable but not significant increase of survival compared to the virus infected, non-treated mice. The addition of human rabies immunoglobulins (HRIG) to the combination in the second experiment almost completely prevented mortality in the pre-exposure treatment group along with a significant reduction of viral titres in the CNS. Post-exposure treatments also greatly improved survival rates. As part of the combination with immunomodulatory compounds, HRIG had a higher impact on survival than alone. In the third experiment the combination was further supplemented with type-I interferons, ribavirin and favipiravir (T-705). As a blood-brain barrier opener, mannitol was also administered. This treatment was unable to prevent lethal consequences of SHBRV-18 infection; furthermore, it caused toxicity in treated mice, presumably due to interaction among the components. In all experiments, viral loads in the CNS were similar in mice that succumbed to rabies regardless of treatment. According to the findings, inhibitors of detrimental host response to rabies combined with antibodies can be considered among the possible therapeutic and post-exposure options in human rabies cases.