Serological evidence suggests that several Simbu serogroup viruses circulated in Israel.

Viruses of the Simbu serogroup are arboviruses that are known to cause outbreaks of abortion, stillbirth and congenitally deformed neonates. This study presents the results of antibody screening of Simbu serogroup viruses in heifers born in Israel after October 2013, and in adult milking cows born before May 2012. Thirteen dairy cattle farms in five regions, and one sheep flock, entered this study. Serum samples that were found to be positive by ELISA were further tested by specific virus- neutralization test against a panel of Simbu serogroup viruses including Akabane, Aino, Sathuperi, Shamonda, and Peaton viruses. Antibody detection in lactating adult cows revealed that several viruses were circulating in Israel between 2008-2014. Moreover, during autumn 2014 the heifers became serum-positive after being exposed to more than one Simbu serogroup virus concurrently. The results of this study shed new light on Simbu virus infections in Israel, and may contribute to the epidemiology of the Simbu serogroup around the Mediterranean Basin in general.

Monitoring for bovine arboviruses in the most southwestern islands in Japan between 1994 and 2014.

BACKGROUND: In Japan, epizootic arboviral infections have severely impacted the livestock industry for a long period. Akabane, Aino, Chuzan, bovine ephemeral fever and Ibaraki viruses have repeatedly caused epizootic abnormal births and febrile illness in the cattle population. In addition, Peaton, Sathuperi, Shamonda and D'Aguilar viruses and epizootic hemorrhagic virus serotype 7 have recently emerged in Japan and are also considered to be involved in abnormal births in cattle. The above-mentioned viruses are hypothesized to circulate in tropical and subtropical Asia year round and to be introduced to temperate East Asia by long-distance aerial dispersal of infected vectors. To watch for arbovirus incursion and assess the possibility of its early warning, monitoring for arboviruses was conducted in the Yaeyama Islands, located at the most southwestern area of Japan, between 1994 and 2014. RESULTS: Blood sampling was conducted once a year, in the autumn, in 40 to 60 healthy cattle from the Yaeyama Islands. Blood samples were tested for arboviruses. A total of 33 arboviruses including Akabane, Peaton, Chuzan, D' Aguilar, Bunyip Creek, Batai and epizootic hemorrhagic viruses were isolated from bovine blood samples. Serological surveillance for the bovine arboviruses associated with cattle diseases in young cattle (ages 6-12 months: had only been alive for one summer) clearly showed their frequent incursion into the Yaeyama Islands. In some cases, the arbovirus incursions could be detected in the Yaeyama Islands prior to their spread to mainland Japan. CONCLUSIONS: We showed that long-term surveillance in the Yaeyama Islands could estimate the activity of bovine arboviruses in neighboring regions and may provide a useful early warning for likely arbovirus infections in Japan. The findings in this study could contribute to the planning of prevention and control for bovine arbovirus infections in Japan and cooperative efforts among neighboring countries in East Asia.

Intraoperative adjustment of eyelid level in aponeurotic blepharoptosis surgery.

There have been numerous reports on the use of aponeurotic surgery to correct involutional blepharoptosis. However, it is still difficult to determine optimal eyelid level during operation. Here we present our new method to adjust eyelid level intraoperatively. After the aponeurosis was temporally sutured to the tarsus, while still in the supine position, the patient was asked to look up, and the position of the eyelid margin was confirmed. The margin should be located above the pupil but within the cornea while the patient gazes up. And it is ideal if the eyelid position is located in the upper half of this range. Although 3 of 29 patients were reoperated on in the follow-up period, only 1 patient required readjustment in the perioperative period. Our method is simple, easy and reduces operative time, because it is not necessary to change patient position during the operation.

Comparison of ultrasonography-assisted closed reduction with conventional closed reduction for the treatment of acute nasal fractures.

Ultrasonography has often been reported to be a useful tool in cases of nasal fracture, not only for diagnosing such fractures but also for intraoperatively assessing surgical outcomes. In this study, we examined the utility of ultrasonography for intraoperatively assessing the results of surgery for acute nasal fractures. In the conventional group, the outcome of each fracture reduction procedure was intraoperatively confirmed by visual inspection and palpation. In the ultrasound group, intraoperative ultrasonography was used to assess the condition of the fracture before and after closed reduction. The outcomes of the reduction procedures and the reoperation rate were compared between the two groups. According to computed tomography-based evaluations, there were no significant differences in the outcomes of the reduction procedures between the two groups (p > 0.05). As for the reoperation rate, two patients (2.8%) in the conventional group underwent reoperations, but no patient (0%) required reoperations in the ultrasound group. However, the difference in the reoperation rate between the two groups was not significant (p > 0.05). These results indicate that visual inspection and palpation are as reliable as ultrasonography for intraoperatively assessing the outcomes of surgery for acute nasal fractures. Surgeons should not depend on ultrasonography alone, but rather should use it in addition to visual inspection and palpation.

Treatment of acute temporomandibular joint dislocation using manipulation technique for disk displacement.

Temporomandibular joint dislocation is not frequently encountered, but it is often difficult to reduce the dislocation with conventional methods described in textbooks. The key points to success of reduction depend on the patient's position, route of approach, and timing of reducing each side. We apply a manipulation technique for disk displacement to the reduction that corresponds to these key points. Using our method, temporomandibular joint dislocation can be easily reduced, without using sedative or analgesics. This method is simple, convenient, and worth trying in place of the conventional method.

Molecular identification of field-collected Culicoides larvae in the southern part of Japan.

Although Culicoides biting midges act as a vector of important human and domestic animal diseases, their ecology is poorly understood. The lack of proper identification systems of Culicoides larvae is one of the main obstacles to progress in research. Based on mitochondrial sequences of 19 Japanese Culicoides species, we designed a universal primer set to amplify the partial sequence of the mitochondrial cytochrome c oxidase I (cox 1). The polymerase chain reaction product amplified from extracted DNA of Culicoides larvae using the primer set was directly sequenced, and species identification based on the variation at cox1 was conducted. Using the molecular identification system, we sorted 243 specimens of field-collected larvae from the southern part of Japan into 10 species including Culicoides arakawae (Arakawa), Culicoides oxystoma Kieffer, and Culicoides brevitarsis Kieffer, which are regarded as vectors of important livestock animal diseases. Eight species of Culicoides larvae, including C. arakawae and C. oxystoma, were recovered from active paddy fields and an abandoned paddy field. The result suggests that paddy fields contribute to breeding a variety of Culicoides species and maintenance and spread of Culicoides-borne pathogens. In contrast, larvae of C. brevitarsis were collected from cattle dung in pastures. The molecular identification system described herein using nucleotide sequences successfully achieved larval identification and will be useful for a better understanding of larval habitats of Culicoides biting midges.

Preauricular transparotid approach to mandibular condylar fractures without dissecting facial nerves.

Preauricular transparotid approach without dissecting the facial nerve was used for surgical treatment of 15 condylar fractures in 14 patients. The parotid fascia was opened just above the fracture site, and by dissecting the parotid gland and masseter muscle, the fracture was directly exposed. The facial nerve itself was not dissected expressly. All fractures could be reduced accurately and fixed firmly with miniplates. A direct approach just above the fracture site provided good vision of the fracture, avoiding facial nerve palsy caused by strong retraction. Moreover, by not dissecting the facial nerve, the operation time was shortened. This approach was useful for surgical treatment of both condylar neck and subcondylar fractures.

Comparison of pediatric and adult nasal fractures.

Nasal fractures are the most common facial fracture in children and adults. Generally, it is believed that reduction of pediatric nasal fracture is more difficult and should be performed earlier compared with that of adult nasal fracture. However, there has been no article to prove this theory. We investigated 423 patients with acute nasal fractures requiring surgery and divided them into the following 2 groups: patients 12 years and younger (pediatric group) and patients 13 years and older (adult group). We then compared these patients in various aspects. There were no significant differences in the cause of fracture or postoperative conditions. Only the type of fracture and the anesthesia were different between these 2 groups. In the pediatric group, the interval between injury and surgery was arbitrarily divided into 2 groups, but there was no significant difference between these groups in the postoperative conditions. Some reports recommended that pediatric nasal fractures should be reduced within 3 to 5 days, but it cannot be proven. In conclusion, it is not necessary to distinguish treatment of pediatric nasal fracture from that of adult nasal fracture.

Genetic reassortment between Sathuperi and Shamonda viruses of the genus Orthobunyavirus in nature: implications for their genetic relationship to Schmallenberg virus.

The recent outbreak of malformations in ruminants in Northern Europe caused by Schmallenberg virus induced us to analyze the genetic properties of the related orthobunyaviruses and clarify their relationship. The sequencing of three genomic RNA segments of Sathuperi, Shamonda and Douglas viruses (SATV, SHAV and DOUV) revealed that the M RNA segment of SATV and DOUV had a high degree of sequence identity with that of Schmallenberg virus, but the S and L RNA segments closely matched those of SHAV. Phylogenetic analysis of the three genomic RNA segments indicated that Schmallenberg virus is a reassortant, with the M RNA segment from SATV and the S and L RNA segments from SHAV.

Treatment of fingertip amputation: comparison of results between microsurgical replantation and pocket principle.

In this article, a comparison of replantation using microsurgical replantation (replantation) and the Brent method and its modification (pocket principle) in the treatment of fingertip amputation is reported. As a classification of amputation level, we used Ishikawa's subzone classification of fingertip amputation, and the cases of amputations only in subzone 2 were included in this study. Between these two groups, there was no statistical difference in survival rate, postoperative atrophy, or postoperative range of motion. In terms of sensory recovery, some records were lost and exact study was difficult. But there was no obvious difference between these cases. In our comparison of microsurgical replantation versus the pocket principle in treatment of subzone 2 fingertip amputation, there was no difference in postoperative results. Each method has pros and cons, and the surgeon should choose which technique to use based on his or her understanding of the characteristics of both methods.

The 2010 foot-and-mouth disease epidemic in Japan.

Foot-and-mouth disease (FMD) occurred recently for the first time in a decade in Japan. The index case was detected on a beef-breeding farm in Miyazaki Prefecture, Southern Japan, on April 20, 2010. After confirmation of this first case, control measures such as stamping out, movement restriction and disinfection were implemented. However, these strategies proved insufficient to prevent the spread of FMD and emergency vaccination was adopted. Up until the last outbreak on July 4, 2010, a total of 292 outbreaks had been confirmed, with about 290,000 animals having been culled. The epidemic occurred in an area with a high density of cattle and pigs, making disease control difficult. Invasion of the disease into a high-density area aided its rapid spread and led to difficulties in locating suitable burial sites. Epidemiological investigations indicated that the disease was introduced into Japan approximately one month before detection. This delay in initial detection is considered to have allowed an increased number of outbreaks in the early stage of the epidemic. Nevertheless, the epidemic was contained within a localized area in Miyazaki Prefecture and was eradicated within three months because of intensive control efforts including emergency vaccination. Although this epidemic devastated the livestock industry in Japan, many lessons can be learnt for the future prevention and control of infectious diseases in animals.

Reduction of nasal fracture using an airway tube as a pivot.

When repairing nasal fracture by closed reduction, an airway tube is useful for the pivot of the forceps. By placing the tube on the upper white lip and using it as a pivot, the fracture is reduced with Walsham forceps or Langenbeck elevator. This method is useful for all nasal fractures that are too firm to reduce by conventional closed reduction.

Differential host gene responses in mice infected with two highly pathogenic avian influenza viruses of subtype H5N1 isolated from wild birds in Thailand.

In Thailand, highly pathogenic avian influenza (HPAI) viruses of subtype H5N1 had been isolated from various wild birds during the HPAI outbreak in poultries. In this study, we examined the pathogenicity of two wild bird isolates (A/Pigeon/Thailand/VSMU-7-NPT/2004; Pigeon04 and A/Tree sparrow/Ratchaburi/VSMU-16-RBR/2005; T.sparrow05) in mice. They showed similar replication in several organs and lethal outcome. However, on day 3 post-infection, Pigeon04 induced mRNA expression of proinflammatory cytokines (IL6 and TNFα) and MIP-2, neutrophil chemoattractant, in the lungs, resulting in severe pneumonia that was accompanied by neutrophil infiltration. In contrast, on day 7 post-infection, T.sparrow05 induced the expression of several cytokines to a greater extent than Pigeon04; it also potently induced mRNA expression of several cytokines in brains of the infected mice that triggered frequent inflammatory events. In sum, our study demonstrated that two HPAI viruses induced different host responses, despite having similar replications, resulting in lethal outcome in mice.

Genetic characterization and susceptibility on poultry and mammal of H7N6 subtype avian influenza virus isolated in Japan in 2009.

From February to March 2009, six strains of H7N6 subtype avian influenza virus were isolated from quails in three farms in Aichi prefecture in Japan. The isolates were shown to be low pathogenic for chicken by the examination performed using the "Manual of Standards for Diagnostic Tests and Vaccines" by World organisation for Animal Health (OIE). The deduced amino acid sequence at the cleavage site was PE (I/Q/L) PKRR (nucleotide sequences were cct gaa (a/c) (t/a) a cc (a/g) aaa aga aga), suggesting persistence in domestic poultry for some time. The direct putative ancestor strain could not be elucidated by phylogenetic analysis of all genome segments of the quail isolates. Diverged date from a putative common ancestor in a non-rooted phylogenetic tree among quail viruses was estimated between March 2002 and July 2004. Three putative N-linked glycosylation sites resided in the vicinity of the receptor binding pocket of HA1 region. They are considered to decrease the reactivity of neutralizing antibody against the virus. Experiments for the infectivity and pathogenicity of a quail strain to poultry indicated that the quail isolate had higher infectivity to quails than chickens and ducks. Direct and dust-borne and/or droplet-borne transmissions among quail were proven in quails with and without direct contact with experimentally infected quails. The virus is seldom transmitted among chickens either directly or indirectly, and indirect transmission from infected quails to chickens was not observed. The pathogenicity of the quail strain for mammalian, pig and mouse was low, although it could replicate in those animals.

Real-time reverse transcription-PCR assay for differentiating the Pandemic H1N1 2009 influenza virus from swine influenza viruses.

Since the Pandemic H1N1 2009 (H1N1pdm) influenza virus emerged in human in 2009, H1N1pdm, classical swine H1, Eurasian avian-like H1, human-like H1 and human-like H3 swine influenza viruses have circulated in pig populations, and avian H9N2 viruses have been isolated in pigs as well. In this study, TaqMan single-step real-time reverse transcription-PCR (rtRT-PCR) assays targeting the hemagglutinin gene were developed to differentiate H1N1pdm from other genetic lineages of the H1 subtype and other subtypes of influenza viruses circulating in human and pig populations for veterinary use. H1N1pdm rtRT-PCR detected H1N1pdm RNA and did not cross-react with classical swine H1, Eurasian avian-like H1, human-like H1, human-like H3 swine and avian H9 influenza viruses RNA. Classical swine H1, Eurasian avian-like H1, human-like H1 and H3 and avian H9 rtRT-PCR were reacted exclusively with viral RNA of their respective lineages and subtypes. The results demonstrate that these assays are useful for the diagnosis of the H1N1pdm virus in both human- and animal-health-related fields.

Genetic characterization of Aino and Peaton virus field isolates reveals a genetic reassortment between these viruses in nature.

Sequence determination and phylogenetic analysis were conducted using the S, M and L RNA segments of the 10 Aino, 6 Peaton and 1 Sango virus (AINOV, PEAV and SANV) field isolates of the genus Orthobunyavirus in the family Bunyaviridae, respectively. The Japanese AINOV strains were genetically stable, but the sequence differences between the Japanese and Australian AINOV strains were considerably larger than those among the Japanese AINOV strains. A similar result was found in the genetic relationship among Japanese and Australian PEAVs, and SANV which was isolated in Nigeria and was thought as a synonym of PEAV, suggesting that geographic separation contributed significantly to the evolution of those viruses. The Australian AINOV strain B7974 is more closely related to the Australian PEAV strain CSIRO110 than to the Japanese AINOV strains in the S and L RNA segments, while the phylogenetic position of the M RNA segment of the B7974 strain was clustered with those of the Japanese AINOV strains. Our findings indicate that the B7974 strain is a reassortment with the M RNA segment derived from AINOV and the S and L RNA segments derived from an Australian PEAV.

Foot-and-mouth disease virus antigen detection enzyme-linked immunosorbent assay using multiserotype-reactive monoclonal antibodies.

Monoclonal antibody (MAb)-based sandwich direct enzyme-linked immunosorbent assay (MSD-ELISA) methods that can detect foot-and-mouth disease virus (FMDV) antigens, both multiserotype (MSD-ELISA/MS) (for O, A, C, and Asia 1) and single-serotype (MSD-ELISA/SS) (for O, A, and Asia 1, specifically), were developed. MAb 1H5 was used as an antigen-trapping antibody that reacted with all seven serotypes of FMDV. The MAbs 71F2, 70C4, 16C6, and 7C2 were used as peroxidase-labeled detecting antibodies for multiple serotypes (O, A, C, and Asia 1), type O, type A, and type Asia 1, respectively, in both MSD-ELISA/MS and MSD-ELISA/SS. Our MSD-ELISAs showed high specificity. They produced a very low background of negative samples (buffer, plasma, and saliva) and were able to detect FMDV antigens from clinical samples (plasma and saliva), with results correlating with those of real-time reverse transcription-PCR. In terms of sensitivity, the MSD-ELISAs showed higher optical density values against each diluted serotype antigen than the indirect sandwich ELISA method, which is currently recommended in the manual of the World Organization for Animal Health. The sensitivity and specificity of the MSD-ELISAs seem to be sufficient for the antigenic diagnosis of FMDV.

Neutralizing monoclonal antibody sandwich liquid-phase blocking enzyme-linked immunosorbent assay for detection of Foot-and-mouth disease virus type O antibodies.

Liquid-phase blocking enzyme-linked immunosorbent assay (LPBE) using the neutralizing monoclonal antibody (mAb) sandwich method (M-LPBE) for detection of Foot-and-mouth disease virus (FMDV) type O antibodies was developed. Two neutralizing mAbs, 72C1 and 65H6, were raised against the FMDV O/JPN/2000 strain, and used as trapping and peroxidase-labeled detecting antibodies, respectively. Sera from animals experimentally infected with FMDV showed specific positive results by M-LPBE, which were correlated with the results of the virus neutralization test (VNT). When 303 negative bovine and 302 negative swine sera were tested, the specificity of M-LPBE was 100% and 99.7%, respectively. In addition, nine samples that had been collected in 2000 in Japan and regarded as evidently false positives by LPBE (supplied by the World Reference Laboratory for Foot-and-Mouth Disease) were uniformly negative by M-LPBE, just like VNT. Therefore, M-LPBE seems to have sufficient specificity for FMDV type O antibody screening and diagnosis.

Phylogenetic relationships of the G gene sequence of bovine ephemeral fever virus isolated in Japan, Taiwan and Australia.

The G gene encoding the neutralization antigen of bovine ephemeral fever virus (BEFV) was characterized in order to define the virus's molecular epidemiology in Japan and the genetic relationships among the Japanese, Taiwanese and Australian isolates. The nucleotide and amino acid sequences of the gene were highly conserved among the Japanese strains, regardless of the year of isolation, and were closely related to the Taiwanese strains. By phylogenetic analysis, the Japanese and Taiwanese strains were classified clearly into three chronological clusters: 1966, 1984-1989 and 1996-2004, indicating that the epidemics of bovine ephemeral fever may occur almost simultaneously in both countries by the same genotype. On the other hand, the Australian strains were distantly related to these East Asian strains and placed in the independent fourth cluster of the phylogenetic tree. It is suggested that three amino acid substitutions at residues 224, 271 and 499 in the neutralizing epitopes, of which two generate new glycosylation sequences, are responsible for antigenic variations of bovine ephemeral fever virus. The cross-neutralization test using the bovine ephemeral fever virus isolated in Japan demonstrated that the vaccine developed based on the oldest Japanese strain, YHL, appears to still be effective for controlling bovine ephemeral fever in Japan.

Comparison of the characters of the plaque-purified viruses from foot-and-mouth disease virus O/JPN/2000.

At least two biotypes were observed at the 2nd passage stage after the isolation of Foot-and-mouth disease Virus (FMDV) O/JPN/2000 strain. These 2 types of viruses differed from their plaque phenotypes and were distinguishable by using a monoclonal antibody (MAb) 64G8 that was made for the FMDV O/JPN/2000 strain. One of these 2 biotypes formed small plaque (SP) and with immuno staining showed a positive reaction to MAb 64G8, while the other formed clear large plaque (LP) and did not react with MAb 64G8. The amino acid sequences of the capsid coding region (VP1-VP4) of the SP virus (SPV) and the LP virus (LPV) revealed two substitutions on the 133rd amino acid in VP2, and the 56th amino acid in VP3. These amino acid changes of SPV and LPV are Asn to Asp, Arg to His, respectively. The Arg of the 56th amino acid in VP3 that have been known as critical position of cell culture adapted virus. Only LPV showed high pathogenicity in suckling mice, and its LD(50) was calculated to be about 10(2) TCID(50)/0.1 ml. These results showed that the SPV that existed at the 2nd passage stage from isolation was a low virulence virus, which may suggest why the pathogenicity of O/JPN/2000 did not show clear symptoms in infected cattle.