Characterisation of the H5 and N1 genes of an Indonesian highly pathogenic Avian Influenza virus isolate by sequencing of multiple clone approach

Hemagglutinin and neuraminidase are the main antigenic determinants of highly pathogenic avian influenza (HPAI) virus. The features of these surface glycoproteins have been intensively studied at the molecular level. The objective of this research was to characterise the genes encoding these glycoproteins by sequencing of multiple clones. The H5 and N1 genes of isolate A/duck/Tangerang/Bbalitvet-ACIAR-TE11/2007 were each amplified in one or two fragments using reverse transcriptase-PCR (RT-PCR), and subsequently cloned into pGEM-T Easy TA cloning system. The sequencing result demonstrated high homology between respective clones but with several variations that were identified as single nucleotide polymorphisms (SNPs). A total of 1,707 base pair and 1,350 base pair of H5 and N1 genes respectively were successfully assembled from multiple clones containing the genes of interest. The features of both H5 and N1 genes from this isolate resemble the typical characteristics of Indonesian strains of H5N1 virus from sub-clade 2.1.3. Key Words: Avian Influenza, Characterization, Gene Cloning, Hemagglutinin, Neuraminidas

Haematology and plasma biochemistry of wild Black flying-foxes, (Pteropus alecto) in Queensland, Australia

This paper establishes reference ranges for hematologic and plasma biochemistry values in wild Black flying-foxes (Pteropus alecto) captured in South East Queensland, Australia. Values were found to be consistent with those of other Pteropus species. Four hundred and forty-seven animals were sampled over 12 months and significant differences were found between age, sex, reproductive and body condition cohorts in the sample population. Mean values for each cohort fell within the determined normal adult reference range, with the exception of elevated levels of alkaline phosphatase in juvenile animals. Hematologic and biochemistry parameters of injured animals showed little or no deviation from the normal reference values for minor injuries, while two animals with more severe injury or abscessation showed leucocytosis, anaemia, thrombocytosis, hyperglobulinemia and hypoalbuminemia

Ανάπτυξη υβριδικών (ανόργανων / οργανικών) ημιαγωγών πολλαπλών στρωμάτων με βάση το CdSe. Μελέτη της δομής και της φωτοηλεκτροχημικής συμπεριφοράς τους

Pteropid bats (flying-foxes) are the natural reservoir of Hendra virus, an emergent paramyxovirus responsible for fatal infection in horses and humans in Australia. Pteropus alecto (the Black flying-fox) and the paraphyletic P. conspicillatus (the Spectacled flying-fox) appear to be the primary reservoir hosts. Previous studies have suggested that physiological and ecological factors may underpin infection dynamics in flying-foxes, and subsequent spillover to horses and in turn humans. We sought to examine temporal trends in urinary cortisol concentration in wild Australian flying-fox populations, to elucidate the putative relationship between Hendra virus infection and physiological stress. Pooled and individual urine samples were non-invasively collected from under roosting flying-foxes at two latitudi-nally disparate regions in the eastern Australian state of Queensland. Hendra virus detection, and (in individual urine samples) sex and species determination were PCR-based. Urinary cortisol measurement used a validated enzyme immunoassay. We found no direct correlation between increased urinary cortisol and Hendra virus excretion, but our findings do suggest a biologically plausible association between low winter temperatures and elevated cortisol levels in P. alecto in the lower latitude Southeast Queensland roosts. We hypothesize an indirect association between low winter temperatures and increased Hendra virus infection and excretion, mediated by the physiological cost of thermoregulation. Our findings and our approach are directly relevant to elaboration of the disease ecology of Nipah virus and other emerging henipaviruses in bats. More broadly, they inform investigation of emerging disease infection dynamics across the wildlife/livestock/human interface

Interaction of sleep and emotion across the menstrual cycle

Menstruating individuals experience an increased risk for sleep and affective disorders, attributed in part to monthly oscillations in sex hormones. Emotional functioning and sleep continuity worsens during the perimenstrual phase of the menstrual cycle. This study examined the interactive effects of sleep, menstrual phase, and emotion in healthy women. Participants (N = 51, 43% Caucasian) aged 18–35 (m = 24 years) completed actigraphy and daily sleep/emotion diaries over two menstrual cycles (m days = 51.29). Diary and actigraphic total wake time at night (TWT) and daily ratings of positive and negative affect were compared across four phases of the menstrual cycle: perimenstrual, mid-follicular, periovulatory, and mid-luteal. Relationships between phase, sleep, and emotion were estimated using multistep hierarchical linear modelling. Mean menstrual cycle length was 28.61 ± 2.69 days. Perimenstrual phase positively predicted anger (p < 0.001) but no other emotions. Additionally, the perimenstrual phase predicted higher rates of TWT, such that diary TWT was 8–16 min longer during the perimenstrual (m = 67.54, SE = 3.37) compared to other phases (p < 0.001). Actigraphic TWT was also increased by 4–7 min (m = 61.54, SE = 3.37) in the perimenstrual phase (p < 0.001). Positive emotions were 0.05–0.10 points lower (p = 0.006–0.02) when TWT was greater in the perimenstrual phase. Greater rates of anger and sleep disruption were seen during the perimenstrual phase compared with other phases. When poor sleep occurred during the perimenstrual phase individuals reported reduced positive emotions. Reducing perimenstrual sleep disruptions may be an important intervention target for those at risk for affective disorders

Degradation and remobilization of endogenous retroviruses by recombination during the earliest stages of a germ-line invasion

Endogenous retroviruses (ERVs) are proviral sequences that result from colonization of the host germ line by exogenous retroviruses. The majority of ERVs represent defective retroviral copies. However, for most ERVs, endogenization occurred millions of years ago, obscuring the stages by which ERVs become defective and the changes in both virus and host important to the process. The koala retrovirus, KoRV, only recently began invading the germ line of the koala (Phascolarctos cinereus), permitting analysis of retroviral endogenization on a prospective basis. Here, we report that recombination with host genomic elements disrupts retroviruses during the earliest stages of germ-line invasion. One type of recombinant, designated recKoRV1, was formed by recombination of KoRV with an older degraded retroelement. Many genomic copies of recKoRV1 were detected across koalas. The prevalence of recKoRV1 was higher in northern than in southern Australian koalas, as is the case for KoRV, with differences in recKoRV1 prevalence, but not KoRV prevalence, between inland and coastal New South Wales. At least 15 additional different recombination events between KoRV and the older endogenous retroelement generated distinct recKoRVs with different geographic distributions. All of the identified recombinant viruses appear to have arisen independently and have highly disrupted ORFs, which suggests that recombination with existing degraded endogenous retroelements may be a means by which replication-competent ERVs that enter the germ line are degraded

Survival of avirulent thermostable Newcastle disease virus (strain I-2) in raw, baked, oiled, and cooked white rice at ambient temperatures

Raw white rice has not been considered a good carrier for oral vaccination, probably because of its antiviral activity. Methods are required to overcome antiviral activity in raw white rice. This study was carried out to determine the effects of various treatments of raw white rice on the survival of strain I-2 of Newcastle disease virus. These included cooking and baking the rice or mixing the rice with vegetable oil prior to coating with vaccine virus. The vaccine-coated rice was then stored for 30 min and 24 h, followed by quantitative recovery of the virus. Thirty min after mixing, uncooked, cooked, and baked rice, and rice mixed with vegetable oil showed titers of 106.2, 107.2, 106.6, and 107.0 EID50/0.1 ml, respectively. After storage for 24 h at 22-25℃, the titers dropped to 105.0, 106.5, 105.0, and 106.0 EID50/0.1 ml for uncooked, cooked, baked, and oiled rice, respectively

Novel insights into viral infection and oncogenesis from Koala Retrovirus (KoRV) infection of HEK293T cells

Koala retrovirus is thought to be an underlying cause of high levels of neoplasia and immunosuppression in koalas. While epidemiology studies suggest a strong link between KoRV and disease it has been difficult to prove causality because of the complex nature of the virus, which exists in both endogenous and exogenous forms. It has been difficult to identify koalas completely free of KoRV, and infection studies in koalas or koala cells are fraught with ethical and technical difficulties, respectively. This study uses KoRV infection of the susceptible human cell line HEK293T and RNAseq to demonstrate gene networks differentially regulated upon KoRV infection. Many of the pathways identified are those associated with viral infection, such as cytokine receptor interactions and interferon signalling pathways, as well as viral oncogenesis pathways. This study provides strong evidence that KoRV does indeed behave similarly to infectious retroviruses in stimulating antiviral and oncogenic cellular responses. In addition, it provides novel insights into KoRV oncogenesis with the identification of a group of histone family genes that are part of several oncogenic pathways as upregulated in KoRV infection

Koala retrovirus viral load and disease burden in distinct northern and southern koala populations

Koala retrovirus (KoRV) displays features of both an endogenous and exogenous virus and is linked to neoplasia and immunosuppression in koalas. This study explores the apparent differences in the nature and impact of KoRV infection between geographically and genetically separated "northern" and "southern" koala populations, by investigating the disease status, completeness of the KoRV genome and the proviral (DNA) and viral (RNA) loads of 71 northern and 97 southern koalas. All northern animals were positive for all KoRV genes (gag, pro-pol and env) in both DNA and RNA forms, whereas many southern animals were missing one or more KoRV genes. There was a significant relationship between the completeness of the KoRV genome and clinical status in this population. The proviral and viral loads of the northern population were significantly higher than those of the southern population (P

Genetic diversity of Koala retrovirus (KoRV) env gene subtypes: Insights into northern and southern koala populations

Koala retrovirus (KoRV) is a recently endogenised retrovirus associated with neoplasia and immunosuppression in koala populations. The virus is known to display sequence variability and to be present at varying prevalence in different populations, with animals in southern Australia displaying lower prevalence and viral loads than northern animals. This study used a PCR and next generation sequencing strategy to examine the diversity of the KoRV env gene in both proviral DNA and viral RNA forms in two distinct populations representative of the “northern” and “southern” koala genotypes. The current study demonstrated that the full range of KoRV subtypes is present across both populations, and in both healthy and sick animals. KoRV-A was the predominant proviral subtype in both populations, but there was marked diversity of DNA and RNA subtypes within individuals. Many of the northern animals displayed a higher RNA viral diversity than evident in their proviral DNA, indicating relatively higher replication efficiency of non-KoRV-A subtypes. The southern animals displayed a lower absolute copy number of KoRV than the northern animals as reported previously and a higher preponderance of KoRV-A in individual animals. These discrepancies in viral replication and diversity remain unexplained but may indicate relative protection of the southern population from KoRV replication due to either viral or host factors and may represent an important protective effect for the host in KoRV’s ongoing entry into the koala genome

Scavenging Ducks and Transmission of Highly Pathogenic Avian Influenza, Java, Indonesia

These ducks may be a source of infection for chickens and humans