Search

Equine herpesvirus-1 (EHV-1) causes respiratory diseases, abortion, and encephalomyelitis in horses. The EHV-1 immediate-early (IE) protein, essential for viral replication, is transactivated by the binding of a multiprotein complex including the open reading frame 12 (ORF12) and some host factors to the IE promoter region. Promoter-associated non-coding RNAs (pancRNAs), which are transcribed from bidirectional promoters, regulate the transcription of neighboring genes in mammals and pathogens. In this study, we identified a novel pancRNA transcribed from across the areas of the 5′-untranslated region and a promoter of EHV-1 IE and named it IE pancRNA. IE pancRNA and mRNA were simultaneously expressed in EHV-1-infected RN33B-A68B2M cells. This pancRNA was also transcribed in RK13 and E. Derm cells, which are highly susceptible to EHV-1 infection. Furthermore, IE pancRNA upregulated IE gene expression in the presence of ORF12, and stable expression of IE pancRNA increased the number of EHV-1-infected RN33B-A68B2M cells. These results suggest that IE pancRNAs facilitate EHV-1 proliferation by promoting IE gene expression.

Mycobacterium avium subsp. hominissuis (MAH) is one of the most prevalent mycobacteria causing non-tuberculous mycobacterial disease in humans and animals. Of note, MAH is a major cause of mycobacterial granulomatous mesenteric lymphadenitis outbreaks in pig populations. To determine the precise source of infection of MAH in a pig farm and to clarify the epidemiological relationship among pig, human and environmental MAH lineages, we collected 50 MAH isolates from pigs reared in Japan and determined draft genome sequences of 30 isolates. A variable number of tandem repeat analysis revealed that most pig MAH isolates in Japan were closely related to North American, European and Russian human isolates but not to those from East Asian human and their residential environments. Historical recombination analysis revealed that most pig isolates could be classified into SC2/4 and SC3, which contain MAH isolated from pig, European human and environmental isolates. Half of the isolates in SC2/4 had many recombination events with MAH lineages isolated from humans in East Asia. To our surprise, four isolates belonged to a new lineage (SC5) in the global MAH population. Members of SC5 had few footprints of inter-lineage recombination in the genome, and carried 80 unique genes, most of which were located on lineage specific-genomic islands. Using unique genetic features, we were able to trace the putative transmission route via their host pigs. Together, we clarify the possibility of species-specificity of MAH in addition to local adaptation. Our results highlight two transmission routes of MAH, one exposure on pig farms from the environment and the other via pig movement. Moreover, our study also warns that the evolution of MAH in pigs is influenced by MAH from patients and their residential environments, even if the MAH are genetically distinct.

Protein arginine methyltransferases (PRMTs) are increasingly recognized as potential therapeutic targets but PRMT7 remains an understudied member of this enzyme family. Here, the authors develop a chemical probe for PRMT7 and apply it to elucidate the role of PRMT7 in the cellular stress response.

Mycobacterium avium subsp. hominissuis (MAH) is one of the most important agents causing non-tuberculosis mycobacterial infection in humans and pigs. There have been advances in genome analysis of MAH from human isolates, but studies of isolates from pigs are limited despite its potential source of infection to human. Here, we obtained 30 draft genome sequences of MAH from pigs reared in Japan. The 30 draft genomes were 4,848,678–5,620,788 bp in length, comprising 4652–5388 coding genes and 46–75 (median: 47) tRNAs. All isolates had restriction modification-associated genes and 185–222 predicted virulence genes. Two isolates had tRNA arrays and one isolate had a clustered regularly interspaced short palindromic repeat (CRISPR) region. Our results will be useful for evaluation of the ecology of MAH by providing a foundation for genome-based epidemiological studies.

This study aimed to follow the time-course pathogenesis of EHV-9 abortion in early and late trimesters. Twenty-seven pregnant hamster dams were divided into three groups: (G1) control, (G2) EHV-9-inoculated on the 5th day (early trimester), and (G3) EHV-9-inoculated on the 10th day of gestation (late trimester). Dams were sacrificed at different time points during gestation and examined for viremia and viral DNA in different fetal and maternal tissues and pathological changes in fetal tissue, placenta, and cytokines. Animals in G3 showed a marked increase in the number of dead fetuses than those in G2. Histopathological findings of G2 showed early band coagulative necrosis of maternal spaces and stromal decidual cells. Necrotic changes were observed within the decidua basalis, spongiotrophoblast layer, and labyrinth. First, the virus was localized within mononuclear leukocytes in the decidua capsularis and basalis, and within the necrotic chorionic villi and cervical epithelium. G3 demonstrated degenerative changes within the chorionic villi and trophospongium. The virus antigen was observed within the chorionic villi, trophoblasts, mononuclear cells, and fetal tissues. In conclusion, EHV-9 induced abortion mostly occurs through necrosis of the chorionic villi and cannot cross through the capsular placenta in the early trimester but can through the developed decidual placentation.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

The grasscutter (also known as the greater cane rat; Thryonomys swinderianus) is a large rodent native to West Africa that is currently under domestication process for meat production. However, little is known about the physiology of this species. In the present study, aiming to provide information about gut microbiota of the grasscutter and better understand its physiology, we investigated the intestinal microbiota of grasscutters and compared it with that of other livestock (cattle, goat, rabbit, and sheep) using 16S rRNA metagenomics analysis. Similar to the other herbivorous animals, bacteria classified as Bacteroidales, Clostridiales, Ruminococcaceae, and Lachnospiraceae were abundant in the microbiome of grasscutters. However, Prevotella and Treponema bacteria, which have fiber fermentation ability, were especially abundant in grasscutters, where the relative abundance of these genera was higher than that in the other animals. The presence of these genera might confer grasscutters the ability to easily breakdown dietary fibers. Diets for grasscutters should be made from ingredients not consumed by humans to avoid competition for resources and the ability to digest fibers may allow the use of fiber-rich feed materials not used by humans. Our findings serve as reference and support future studies on changes in the gut microbiota of the grasscutter as domestication progresses in order to establish appropriate feeding methods and captivity conditions.

Chlamydia-related bacteria of the Chlamydiales order have recently been described as emerging pathogens that cause pneumonia and abortion in animals and humans. We investigated the presence of Chlamydiales using real-time polymerase chain reaction (PCR) by targeting the 16S rRNA gene of a broad range of Chlamydiales in 827 fecal samples from pet birds kept in individual homes in Japan. Of the 827 samples, 493 (59.6%) tested positive for the Chlamydiales 16S rRNA gene in the real-time PCR assay. We determined the nucleic acid sequences of PCR products from 17 Chlamydiales strains. A homology search and phylogenetic analysis using these sequences confirmed that the detected Chlamydiales included C. pecorum and a broad range of Chlamydiarelated bacteria. To the best of our knowledge, this is the first study to detect a wide range of Chlamydia-related bacteria in birds. [ABSTRACT FROM AUTHOR]

The members of family Chlamydiaceae have a broad host range and cause many kinds of diseases in humans and animals. Several cases of Chlamydiaceae being detected in atypical hosts have been reported recently. Consequently, cross-species monitoring of Chlamydia in wildlife and livestock is pertinent for public health, animal hygiene and wildlife conservation. In this study, we conducted molecular surveillance of Chlamydia in wild birds and livestock around a small village in the foothills of Mt. Afadjato, Ghana where direct contact between wildlife and livestock occurs. Among 29 captured wild birds and 63 livestock, 5 sheep, 30 goats and 28 chickens, the positive ratios of Chlamydia were 24.1%, 40.0%, 43.3% and 26.9%, respectively. Chlamydia pecorum was detected in wild birds, goats, sheep and chickens. On the basis of the variable domain 2 region of ompA, several samples from different hosts showed identical sequences and were phylogenetically located to the same clusters. In addition, using ompA, C. psittaci, C. abortus and C. gallinacea were also detected in this small habitat. Further genetic and pathogenic analyses of the chlamydial distribution in this area, which represents the interface of wild and domestic animal interactions, may improve our knowledge of their transmission among different hosts. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the neuropathogenesis of equine herpes virus 9 (EHV-9) by studying the effects of a single point mutation introduced in two different EHV-9 genes. The two EHV-9 mutants, 14R and 19R, were generated carrying a point mutation in two separate EHV-9 genes. These mutants, along with the wild-type EHV-9, were used to infect a hamster model. The EHV-9- and 19R-infected groups showed earlier and more severe clinical signs of infection than the 14R-infected group. The white blood cells (WBCs) count was significantly increased in both EHV-9- and 19R-infected groups compared to the 14R-infected group at the 4th day post infection (DPI). Viremia was also detected earlier in both EHV-9- and 19R-infected groups than 14R-infected group. There were differences in the anterograde transmission pattern of both EHV-9 and 19R compared to 14R inside the brain. Serum TNF-α, IL-6 and IFN-γ levels were significantly increased in both EHV-9- and 19R-infected groups compared to the 14R-infected group. Histopathological and immunohistochemical analyses revealed that the mean group scores for the entire brain were significantly higher in both EHV-9- and 19R- infected groups than 14R-infected group. Collectively, these results confirm that the gene product of Open Reading Frame 19 (ORF19) plays an important role in EHV-9 neuropathogenicity and that the mutation in ORF19 is responsible for the attenuation of EHV-9. [ABSTRACT FROM AUTHOR]

Equine herpesvirus type 4 (EHV-4) is one of the most important pathogens in horses. To clarify the key genes of the EHV-4 genome that cause abortion in female horses, we determined the whole genome sequences of a laboratory strain and 7 Japanese EHV-4 isolates that were isolated from 2 aborted fetuses and nasal swabs of 5 horses with respiratory disease. The full genome sequences and predicted amino acid sequences of each gene of these isolates were compared with of the reference EHV-4 strain NS80567 and Australian isolates that were reported in 2015. The EHV-4 isolates clustered in 2 groups which did not reflect their pathogenicity. A comparison of the predicted amino acid sequences of the genes did not reveal any genes that were associated with EHV-4-induced abortion. [ABSTRACT FROM AUTHOR]

The changes in protein degradation behavior in fish surimi upon addition of different protease inhibitors were examined, with the aim of identifying the proteases involved in the modori reaction in different species and grades of fish and at different temperatures. The modori reactions at 40, 50, and 60°C in four frozen surimi samples: grades FA and KA blue grenadier and grades SA and RA Alaska pollock were investigated. In the grade FA blue grenadier surimi, the modori reaction primarily involved metalloproteases at 40°C, serine proteases and cysteine proteases at 50°C, and cysteine proteases at 60°C. The reaction in the grade KA blue grenadier surimi involved multiple proteases at 40°C, and metalloproteases and cysteine proteases at 50 and 60°C. In the Alaska pollock surimi, multiple proteases were suggested to be equally involved in the modori reaction at 40, 50, and 60°C in both grades SA and RA samples.

The compositions and inosinic acid contents of ATP-related compounds in various processed fish meat products were examined in this study. Moreover, the relationships between these variables and the heating processes used on the products were investigated. Dried and salted fish meat products manufactured immediately after manufacturing, were found to maintain some inosinic acid contents, whereas those stored for a long period of time had low inosinic acid contents. It was observed that the inosine content in processed fish meat products manufactured without the heat treatment was decreased during the storage at 5°C. Therefore, the low inosinic acid content in such processed fish meat products was considered to be because the maintenance of inosine monophosphate (IMP)-degrading enzyme activity, resulting in the decomposition of inosinic acid even under refrigerated storage. The inosine content of unheated water extract decreased faster than that of fish meat product, and the effect of heating temperature could be observed more clearly. The results suggested that IMP-degrading enzymes in club mackerel, Atlantic bluefin tuna, and Japanese jack mackerel were deactivated by heating at 70°C or higher, while those in fine patterned puffer and half-smooth golden pufferfish were deactivated with heating at 60°C or higher. Since IMP decomposition was suppressed incrementally with heating, it appears that the multiple enzymes involved in the decomposition of inosinic acid might have varying thermal stabilities.

Avian bornaviruses (ABVs) were recently discovered as the causative agents of proventricular dilatation disease (PDD). Although molecular epidemiological studies revealed that ABVs exist in Japan, no Japanese isolate has been reported thus far. In this study, we isolated four strains of Psittaciform 1 bornavirus from psittacine birds affected by PDD using QT6 quail cells. To our knowledge, this is the first report to isolate ABVs in Japan and to show that QT6 cells are available for ABV isolation. These isolates and QT6 cells would be powerful tools for elucidating the fundamental biology and pathogenicity of ABVs. [ABSTRACT FROM AUTHOR]

In the previous study, one of the authors investigated stationary response characteristics of linear systems subjected to non-Gaussian random excitation. It was revealed that the probability distribution of the stationary response varies significantly depending upon the bandwidth of the excitation power spectrum. In real engineering problems, on the other hand, the transient response before reaching the stationary state is also crucial. Therefore, in this paper, we explore transient response characteristics to non-Gaussian random excitation by Monte Carlo simulation. The non-Gaussian excitation is prescribed by both a probability density function and a power spectrum with bandwidth parameter. In order to find the response characteristics that appears commonly for various non-Gaussian excitation, we consider four kinds of non-Gaussian distributions for the probability distributions of the excitations. Their shapes are much different from a Gaussian distribution and also distinct from each other. For such non-Gaussian random excitations, we look into the relationship between the transient response characteristics and the excitation bandwidth. Specifically, first, the transient response distribution is examined. Then, we observe the variance, skewness and kurtosis of the transient response to discuss the time variation of the response distribution in more detail.

VP22 is a major tegument protein of Equine herpesvirus type 1 (EHV-1) that is a conserved protein among alphaherpesviruses. However, the roles of VP22 differ among each virus, and the roles of EHV-1 VP22 are still unclear. Here, we constructed an EHV-1 VP22 deletion mutant and a revertant virus to clarify the role of VP22. We found that EHV-1 VP22 was required for efficient viral growth in cultured cells, but not for virulence in a hamster model. [ABSTRACT FROM AUTHOR]

The article presents a study which suggests that Chlamydia pecorum strains similar to 22-58 and 24-100 might be endemic in Yamaguchi Prefecture, Japan and cause enteric disease in cattle. Topics include the isolation of C. pecorum (22-58) from the jejunum of a calf which died of necrotizing enterocolitis in 2010, the isolation of C. pecorum (24-100) from the feces of a calf with diarrhea in 2012, and the significant increase in neutralizing antibody titers against C. percorum in paired sera.

This study investigated possible differences in the suwari reaction among surimi and examined the possibility of using measurements obtained during the analysis of the suwari reaction as measures of quality. The changes in breaking strength during the suwari reaction of six kinds of frozen surimi, comprising high-grade and low-grade samples of three species – blue grenadier, Alaska pollock, and threadfin bream – were examined to determine the setting speed and activation energy. There was little difference in the setting speed between the grades of each fish species, likely due to the lower setting ability of the low-grade surimi which quickly reached the maximum breaking strength. In contrast, the activation energy of the high-grade surimi samples of all three species was lower by 1.3–2.5 times. Thus, the activation energy of the suwari reaction was demonstrated to be an effective measure of surimi setting ability and quality, which varies between species and surimi grade.

The article discusses a study that investigated the subclinical infection rate of psittacine beak and feather disease (PBFD) in live psittacine birds imported from some countries and bred in Japan from April 2003 to March 2004. An overview of PBFD is presented. It also discusses the process of randomly selecting beak and feather disease virus (BFDV) samples for sequencing, and the details relating to the nucleotide sequences of the selected BFDV samples.

Abstract  Infectious bursal disease (IBD) virus (IBDV) serotype 1 is the causative agent of IBD, a highly contagious immunosuppressive disease of young chickens. In this study, we examined IBDV infection in apparently healthy 21 guinea fowls and 20 pigeons obtained in Tanzania and Zambia by virus neutralization test (VNT) and reverse transcription polymerase chain reaction (RT-PCR) for the VP2 hypervariable region (VP2-HVR) of IBDV. Two guinea fowls (9.5%) in Tanzania were RT-PCR and VNT positive for IBDV, and 1 pigeon (5%) in Tanzania was RT-PCR positive and VNT negative. Phylogenetic analysis based on the nucleotide sequences of the PCR products indicated that segment A of IBDV detected from one guinea fowl and a pigeon belonged to the very virulent genotype of European/Asian type, while the other IBDV detected from a guinea fowl belonged to the classical genotype. To our knowledge, this is the first report of detection of the IBDV genome in free-living pigeons and guinea fowls. The detection of IBDV from apparently healthy guinea fowls and pigeons elucidates the role of wild birds in the epidemiology of IBDV. [ABSTRACT FROM AUTHOR]

Objectives: Chlamydiae are obligate intracellular bacteria, causing a variety of diseases, i.e. pneumonia, sexually transmitted disease, conjunctivitis and zoonosis. Tryptophan depletion by interferon-gamma (IFN-γ) is the most important host defence system against chlamydial infection. Thus chlamydial tryptophan metabolism is thought to play key roles for IFN-γ resistance, persistent infection and host/tissue tropisms. We tested tryptophan derivatives for activity against chlamydia-infected cells.Methods: Rates of chlamydial infection and sizes of the inclusions were evaluated by in vitro infection using three Chlamydiaceae species, Chlamydia trachomatis, Chlamydophila pneumoniae and Chlamydophila felis, which show significant divergence of tryptophan synthesis genes and different susceptibilities to IFN-γ.Results: Melatonin and serotonin, which are recognized as neural hormones for maintenance of organism homeostasis, reduced chlamydial infection but not other bacterial growth tested here. Unlike IFN-γ, melatonin limited infection of all three chlamydiae and the effects were not recovered by tryptophan supplementation. Melatonin treatment only of host cells could diminish infection and the infection reduction was neutralized by a pertussis toxin, an inhibitor of G proteins. Ligands of melatonin and serotonin receptors also hampered infection.Conclusions: Inhibition mechanisms of chlamydial infection by melatonin and serotonin appear to be different from those of IFN-γ and involve specific G-protein-coupled receptors. Melatonin is deemed to inhibit early progression of the chlamydial development cycle, such as establishment of intracellular infection and/or conversion from elementary body to reticulate body. Utilization of melatonin, serotonin or their derivatives may be advantageous for harmless prevention of chlamydial infection. [ABSTRACT FROM AUTHOR]

Twenty-three clinical strains of Chlamydia trachomatis were isolated from men with chlamydial nongonococcal urethritis and examined for the presence of fluoroquinolone resistance-associated alterations in GyrA and ParC. The minimum inhibitory concentrations (MICs) of fluoroquinolones, erythromycin, and tetracycline were determined for 6 of the 23 isolates. In 12 of the 23 isolates, a single amino-acid change was found in GyrA, and in 1 isolate, two amino acids were changed. In all 23 isolates, an Arg-83-to-Gly substitution was observed in ParC, and in 3 isolates, an additional amino-acid change was found. Some changes occurred within the quinolone resistance-determining regions (QRDRs) of GyrA and ParC, but not at positions critical for fluoroquinolone resistance. Of the 6 isolates for which MICs of the agents were determined, 1 isolate had a Cys-66 ? Arg substitution in GyrA, and all had the Arg-83 ? Gly substitution in ParC. However, all 6 isolates were susceptible to fluoroquinolones. First-pass urine specimens were obtained from two men who were positive for C. trachomatis after levofloxacin treatment, and the gyrA and parC genes of C. trachomatis were amplified by polymerase chain reaction (PCR) and examined for fluoroquinolone resistance-associated mutations. Pre- and post-treatment C. trachomatis persisting in each of them had identical amino-acid sequences in the QRDR of GyrA and ParC. Further, the substitutions found in GyrA and ParC were not located at positions critical for fluoroquinolone resistance. The present study suggests that fluoroquinolone resistance-associated alterations in GyrA and ParC may be uncommon in clinical strains of C. trachomatis. [ABSTRACT FROM AUTHOR]

Fish express two different chitinases, acidic fish chitinase-1 (AFCase-1) and acidic fish chitinase-2 (AFCase-2), in the stomach. AFCase-1 and AFCase-2 have different degradation patterns, as fish efficiently degrade chitin ingested as food. For a comparison with the enzymatic properties and the primary structures of chitinase isozymes obtained previously from the stomach of demersal fish, in this study, we purified chitinase isozymes from the stomach of Japanese sardine Sardinops melanostictus, a surface fish that feeds on plankton, characterized the properties of these isozymes, and cloned the cDNAs encoding chitinases. We also predicted 3D structure models using the primary structures of S. melanostictus stomach chitinases. Two chitinase isozymes, SmeChiA (45 kDa) and SmeChiB (56 kDa), were purified from the stomach of S. melanostictus. Moreover, two cDNAs, SmeChi-1 encoding SmeChiA, and SmeChi-2 encoding SmeChiB were cloned. The linker regions of the deduced amino acid sequences of SmeChi-1 and SmeChi-2 (SmeChi-1 and SmeChi-2) are the longest among the fish stomach chitinases. In the cleavage pattern groups toward short substrates and the phylogenetic tree analysis, SmeChi-1 and SmeChi-2 were classified into AFCase-1 and AFCase-2, respectively. SmeChi-1 and SmeChi-2 had catalytic domains that consisted of a TIM-barrel (β/α)8–fold structure and a deep substrate-binding cleft. This is the first study showing the 3D structure models of fish stomach chitinases.

Nucleotide sequences of the VP2 hypervariable region (VP2-HVR) of 10 infectious bursal disease viruses detected in indigenous and exotic chickens in Zambia from 2004 to 2005 were determined. Phylogenetic analysis showed that the viruses diverged into two genotypes and belonged to the African very virulent types (VV1 and VV2). In the phylogenetic tree, strains in one genotype clustered in a distinct group and were closely related to some strains isolated in western Africa (VV1), with nucleotide similarities of 95.7% – 96.5%. Strains in the other genotype were clustered within the eastern African VV type (VV2), with nucleotide similarities of 97.3% – 98.5%. Both genotypes were distributed in the southern parts of Zambia and had a unique conserved amino acid substitution at 300 (E→A) in addition to the putative virulence marker at positions 222(A), 242(I), 256(I), 294(I) and 299(S). These findings represent the first documentation of the existence of the African VV-IBDV variants in both indigenous and exotic chickens in Zambia. [ABSTRACT FROM AUTHOR]