Fine mapping epitope on glycoprotein Gc from Crimean-Congo hemorrhagic fever virus

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonosis, caused by CCHF virus (CCHFV) and which there are no diagnostic or therapeutic strategies. The C-terminus of glycoprotein (Gc) encoded by the CCHFV M gene is responsible for CCHFV binding to cellular receptors and acts as a neutralizing-antibody target. In this study, a modified biosynthetic peptide technique (BSP) was used to identify fine epitopes of Gc from the CCHFV YL04057 strain using rabbit antiserum against CCHFV-Gc. Six B cell epitopes (BCEs) and one antigenic peptide (AP) were identified: E1 (⁸⁸VEDASES⁹⁴), E2 (¹¹⁷GDRQVEE¹²³), E3 (²⁴¹EIVTLH²⁴⁶), AP-4 (²⁸¹DFQVYHVGNLLRGDKV²⁹⁶), E5a (³⁷⁰GDTP QLDL³⁷⁷), E5b (³⁷³PQLDLKAR³⁸⁰), and E6 (⁴⁴³HVRSSD⁴⁴⁸). Western blotting analysis showed that each epitope interacted with the positive serum of sheep that had been naturally infected with CCHFV, and the results were consistent with that of Dot-ELISA. The multiple sequence alignment (MSA) revealed high conservation of the identified epitopes among ten CCHFV strains from different areas, except for epitopes AP-4 and E6. Furthermore, three-dimensional structural modeling showed that all identified epitopes were located on the surface of the Gc “head” domain. These mapped epitopes of the CCHFV Gc would provide a basis for further increase our understanding CCHFV glycoprotein function and the development of a CCHFV epitope-based diagnostics vaccine and detection antigen.     

Recent progress in molecular biology of Crimean-Congo hemorrhagic fever

Crimean–Congo hemorrhagic fever (CCHF) is a severe hemorrhagic fever in humans with a case fatality rate of up to 50%. A causative agent of CCHF is CCHF virus, which is a tick-borne virus in the family Bunyaviridae, genus Nairovirus. The virus is transmitted to humans through infected tick bites, squashed ticks or from direct contact with viremic animals or humans. Outbreaks of CCHF have been documented in Africa, the Middle East, Eastern Europe and Western Asia where the vector and/or reservoir ticks of Hyalomma spp. are distributed. Recent advances in molecular and biochemical analyses of CCHF virus revealed that the virus encodes larger proteins compared to other genus of Bunyavirus and the processing of viral proteins are complicated. Recent studies also showed that the CCHF viruses are relatively divergent in its genome sequence and the viruses are grouped in seven different clades. In general, these phylogenetic analyses based on sequences of S-RNA and L-RNA segment of CCHF viruses indicate that the seven clades correlate with their geographical location. The phylogenetic topology based on M-RNA segment sequences of CCHF viruses is different from those based on S-RNA and L-RNA segments. These analyses indicate that M-RNA segment reassortment events occur more frequently than those in S- and L-RNA segments.     

An antigenic investigation of Crimean-Congo hemorrhagic fever virus (CCHFV) in hard ticks from provinces in northern Turkey

Crimean-Congo hemorrhagic fever virus threatens human health. Exposure of the infected tick is a strong risk factor for human disease. In this study, the hard ticks collected from a variety of mammalian species (cattle, sheep, goat, and buffalo) and a turtle in either coastal or inland Black Sea region of Turkey were surveyed for the presence of antigen from Crimean-Congo hemorrhagic fever virus (CCHFV). CCHFV antigen was found in 46 of 421 tick pools (10.92%). Positivity rates for the provinces varied and were as follows: Samsun 33.87%, Ordu 4.34%, Giresun 8.86%, Sinop 6.09%, Amasya 7.40%, Tokat 5.08%, and Sivas 8.06%. CCHFV antigen was detected in seven of 11 tick species tested. These results suggest that these hard ticks may act as a reservoir for CCHFV in northern Turkey.     

Survey of ixodid tick species on domestic cats in Japan

Various species of ixodid ticks, attached to domestic cats in Japan, were identified in spring (April to June) and autumn (September to November). In the spring, a total of 282 ticks, including 61 larvae, 70 nymphs, 127 females and 24 males were collected from 126 cats. Of these, 264 were identified up to the species level. In the spring, Haemaphysalis longicornis was the most frequently (39.7%, 50/126) found tick species on feline hosts, followed by Ixodes ovatus (35.0%, 44/126), Ixodes nipponensis (15.9%, 20/126) and Haemaphysalis flava (9.5%, 12/126). Small numbers of Haemaphysalis megaspinosa, Haemaphysalis japonica, Ixodes persulcatus, Ixodes granulatus and Amblyomma testudinarium were also recovered. H. longicornis was the most frequently found tick species on cats around riversides or river basins, while I. ovatus and I. nipponensis were more frequently found on cats kept near woodland or related areas. I. nipponensis was more frequently found on castrated males. No major statistical differences in the frequency of tick attachment among sex, age or hair length for the three major tick species were found. Of 205 ticks including 173 (84.4%) larvae, 27 (13.2%) nymphs, 4 (2.0%) females and 1 (0.5%) male recovered from 62 cats in autumn, only 32 (15.6%) were identified. Most of the larvae were fully- or partly-engorged Haemaphysalis spp., and it was difficult to identify them further by morphological characterization.     

A climate-based model for the development of the ixodid tick, Rhipicephalus appendiculatus in East Coast fever zones

East Coast fever (Theileria parva infection) is an important parasitic disease of cattle in East and Central Africa. Past experimental studies have provided a great deal of information on the dynamics of the life cycle of the tick vector, Rhipicephalus appendiculatus. This paper describes a mathematical model based on field observations to explain the close relationship between the tick life cycle and climate. The model provides a basis for the future study of different ECF control programmes using computer experiments.     

Montenegro's skin reactions and antibodies against different Leishmania species in dogs from a visceral leishmaniosis endemic area

In this study, humoral (circulating anti-Leishmania antibodies) and cellular (Montenegro's skin test) immune responses of dogs from an endemic area of visceral leishmaniosis were tested using Leishmania chagasi, Leishmania amazonensis and Leishmania braziliensis antigens. The antibody response was tested in three animal groups, selected according to their anti-L. chagasi antibody activity, as measured by ELISA in the serum: 19 negative (O.D. below 0.30), seven with undefined (O.D. between 0.40 and 0.70) and 12 positive (O.D. above 1.0) ELISA result. In the group of animals with positive ELISA, the antibody activity against L. chagasi antigens (mean O.D.=1.31) was significantly higher (ANOVA, P<0.01) than against L. amazonensis (mean O.D.=0.88) or L. braziliensis (mean O.D.=0.87) antigens. The Montenegro's skin test results obtained with L. chagasi and L. braziliensis antigens showed a fair agreement (kappa=0.309). The same was observed when antigens from L. braziliensis and L. amazonensis were compared (kappa=0.374), whereas a moderate agreement between the results from tests performed with L. chagasi and L. amazonensis antigens was observed (kappa=0.530). The induration areas obtained with L. braziliensis antigen were smaller than those obtained with the other antigens. The data presented herein indicate that the use of antigens from different Leishmania species may interfere with the results of the immunological tests performed in dogs in an endemic area of visceral leishmaniosis.     

A survey of cattle producers in the Boophilus microplus endemic area of Queensland to determine attitudes to the control of and vaccination against tick fever

SUMMARY: A survey by mail was used to determine the views of beef producers in the Boophilus microplus endemic area of Queensland on the control of and vaccination against tick fever. Data from 448 questionnaires were analysed, representing 2.7% of beef producers in the survey area. Producers considered buffalo fly ( Haematobia irritans exigua ) infestation as the most important problem whereas tick fever ranked sixth overall. Private veterinarians were regarded as the most important source of information on vaccines for cattle followed by a weekly rural newspaper. From the survey we estimate that about 33% of producers used the tick fever vaccine produced by the Tick Fever Research Centre of Queensland Department of Primary Industries but there were significant (P < 0.05) variations between regions and herds. Large herds (≥ 400 head) in south-east Queensland were the most likely to be vaccinated against tick fever. Of the producers who did not use the vaccine, over 70% replied that there was no need to vaccinate because of the low risk of the disease in their herds. In 52% of unvaccinated herds the treatment of animals with acaricide was considered the most important means of tick fever control and 61% of these herds comprised Bos indicus x Bos taurus or Bos indicus cattle.     

Serologic and molecular survey of horses to Coxiella burnetii in East of Iran a highly endemic area

There are few reports about Q fever in horse populations worldwide. This study aimed to detect the C. burnetii infection by serologic and molecular confirmation using commercial ELISA kit and real-time PCR in the East of Iran a region highly endemic. A total of 177 blood samples and 115 vaginal swabs were randomly collected from horses in East of Iran. The sera samples were analyzed for anti C.burnetii Ig G antibodies by a commercial ELISA kit and nucleic acid extraxted from vaginal samples were used to determine the C. burnetii DNA by real-time PCR assay. Antibodies were detected in 5.64 % (10/177) of sera samples and C. burnetii DNA was detected in 7.82 % (9/115) of horse vaginal samples. There was no significant difference in seroprevalence in different sex, age and breed groups. Our study showed that horses could be considered as a mild potential reservoir of C. burnetii which may be effective on horse health status. However, additional studies are needed to assess whether the horse could be considered as a relevant transmission risk indicator for Q fever.     

Analysis of classical swine fever virus replication kinetics allows differentiation of highly virulent from avirulent strains

To study the replication of classical swine fever virus (CSFV) in cell culture, kinetics of viral plus-strand RNA synthesis, of viral structural and non-structural protein expression as well as of secreted and cell-associated infectious virus were determined. Highly virulent, moderately virulent and avirulent strains that were tested in standardized animal experiments to confirm their virulence were used to search for in vitro parameters allowing the differentiation of strains according to their virulence. No significant qualitative or quantitative differences were found between the strains studied when either RNA replication or protein synthesis were investigated. However, the ratio of cell-associated virus versus secreted virus proved to be considerably lower for the highly virulent strains when compared to avirulent or moderately virulent strains. These data suggest that highly virulent strains of CSFV can be distinguished in cell culture from strains with reduced virulence.     

Comparison of virulence of various hantaviruses related to hemorrhagic fever with renal syndrome in newborn mouse model

The virulence of hantaviruses that are antigenically related but have different genetic characteristics from the prototype of hantavirus, Hantaan (HTN) virus, was examined in newborn mice. The H5 and B78 strains of the Amur (AMR) genotype, the Bao14 strain of the Far East (FE) genotype, and the 76-118 strain of HTN virus were inoculated subcutaneously (1focus-forming unit; FFU) into newborn mice. All of the AMR and FE genotype viruses inoculated mice were died by 16 days post-infection (dpi) and 21 dpi, respectively, while 50% of the HTN virus inoculated mice survived until 30 dpi. The AMR and FE genotype viruses inoculated mice had high viral titers in the lung (1.3x10(6) to 1.3x10(8) FFU/gram [g] tissue) , brain (2.1x10(7) to 1.2x10(9) FFU/g tissue), and kidney(2.5x10(5) to 1.6x10(7) FFU/g tissue), and showed a detectable level of antibodies (titers 1:16-1:32) at 14 dpi. In contrast, the HTN virus infected mice had viruses only in the lungs at low titers (1.1-5.3x10(5) FFU/g tissue). Observations of body-weight changes revealed that the AMR and FE genotype viruses inoculated mice had lower growth rates than the HTN virus inoculated mice. These data suggest that the AMR and FE genotype viruses are more virulent than the HTN virus in newborn mice.     

Effects of tick infestation on Boran (Bos indicus) cattle immunised against theileriosis in an endemic area of Kenya

Boran (Bos indicus) heifers were immunised by infection with local Theileria parva parva, T p lawrencei and T mutans stocks and treated with parvaquone and later exposed to natural tick and tick-borne disease challenge in the Trans-Mara Division of Kenya. The Theileria species parasites in the challenge were maintained in African buffalo and cattle and the tick vectors were supported by several species of wild Bovidae and domestic livestock present in the area. Thirty immune cattle were observed for 30 weeks while grazing on a ranch in the Trans-Mara Division. Of these, 15 were immersed in toxaphene at weekly or twice-weekly intervals while 15 cattle remained without acaricide application. Cattle which became pregnant were withdrawn from the experiment. There was no evidence of any clinical tick-borne disease in either group of cattle during the experiment. Five species of ixodid ticks infested the cattle during the experiment and cattle which were not treated with acaricide had far more ticks. Rhipicephalus appendiculatus was the most abundant tick species, with a mean infestation of 46 adults per animal in the undipped cattle. This tick also appeared to be the cause of the observed reduction in weight gains. Major haematological parameters did not differ significantly between the groups. Behavioural studies showed that the undipped cattle spent less time grazing and ruminating. This study has shown that, at the expense of some loss in productivity, zebu cattle, immunised against ticks and theileriosis, can be kept despite tick infestation.     

Role of splenectomy in prevention of hemorrhagic enteritis and death from hemorrhagic enteritis virus in turkeys

Hemorrhagic diarrhea, gross hemorrhagic enteritis, and death caused by intravenous virus injection of hemorrhagic enteritis virus (HEV) were prevented in otherwise susceptible turkey poults by surgical splenectomy. The splenectomized poults produced anti-HEV antibodies, which indicated that splenectomy did not completely prevent replication of the virus. These results indicate that the spleen is necessary for the development of the intestinal lesions of this disease. The role of a toxic factor in this disease is discussed.