Advances in Virology The latest articles from Hindawi © 2018 , Hindawi Limited . All rights reserved. Adaptation and Molecular Characterization of Two Malaysian Very Virulent Infectious Bursal Disease Virus Isolates Adapted in BGM-70 Cell Line Sun, 05 Nov 2017 00:00:00 +0000 Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 109.50 TCID50/mL and  TCID50/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogenetic analysis showed that these two isolates were of the vvIBDV. It appears that a single mutation of UPM190 and UPM0081 IBDV isolates at D279N could facilitate vvIBDV strain adaptability in CEE and BGM-70 cultures. Nafi’u Lawal, Mohd Hair-Bejo, Siti Suri Arshad, Abdul Rahman Omar, and Aini Ideris Copyright © 2017 Nafi’u Lawal et al. All rights reserved. Phage-Displayed Peptides Selected to Bind Envelope Glycoprotein Show Antiviral Activity against Dengue Virus Serotype 2 Sun, 10 Sep 2017 00:00:00 +0000 Dengue virus is a growing public health threat that affects hundreds of million peoples every year and leave huge economic and social damage. The virus is transmitted by mosquitoes and the incidence of the disease is increasing, among other causes, due to the geographical expansion of the vector’s range and the lack of effectiveness in public health interventions in most prevalent countries. So far, no highly effective vaccine or antiviral has been developed for this virus. Here we employed phage display technology to identify peptides able to block the DENV2. A random peptide library presented in M13 phages was screened with recombinant dengue envelope and its fragment domain III. After four rounds of panning, several binding peptides were identified, synthesized, and tested against the virus. Three peptides were able to block the infectivity of the virus while not being toxic to the target cells. Blind docking simulations were done to investigate the possible mode of binding, showing that all peptides appear to bind domain III of the protein and may be mostly stabilized by hydrophobic interactions. These results are relevant to the development of novel therapeutics against this important virus. Carolina de la Guardia, Mario Quijada, and Ricardo Lleonart Copyright © 2017 Carolina de la Guardia et al. All rights reserved. Evaluation of NxTAG Respiratory Pathogen Panel and Comparison with xTAG Respiratory Viral Panel Fast v2 and Film Array Respiratory Panel for Detecting Respiratory Pathogens in Nasopharyngeal Aspirates and Swine/Avian-Origin Influenza A Subtypes in Culture Isolates Tue, 29 Aug 2017 00:00:00 +0000 This study evaluated a new multiplex kit, Luminex NxTAG Respiratory Pathogen Panel, for respiratory pathogens and compared it with xTAG RVP Fast v2 and FilmArray Respiratory Panel using nasopharyngeal aspirate specimens and culture isolates of different swine/avian-origin influenza A subtypes (H2N2, H5N1, H7N9, H5N6, and H9N2). NxTAG RPP gave sensitivity of 95.2%, specificity of 99.6%, PPV of 93.5%, and NPV of 99.7%. NxTAG RPP, xTAG RVP, and FilmArray RP had highly concordant performance among each other for the detection of respiratory pathogens. The mean analytic sensitivity (TCID50/ml) of NxTAG RPP, xTAG RVP, and FilmArray RP for detection of swine/avian-origin influenza A subtype isolates was 0.7, 41.8, and 0.8, respectively. All three multiplex assays correctly typed and genotyped the influenza viruses, except for NxTAG RRP that could not distinguish H3N2 from H3N2v. Further investigation should be performed if H3N2v is suspected to be the cause of disease. Sensitive and specific laboratory diagnosis of all influenza A viruses subtypes is especially essential in certain epidemic regions, such as Southeast Asia. The results of this study should help clinical laboratory professionals to be aware of the different performances of commercially available molecular multiplex RT-PCR assays that are commonly adopted in many clinical diagnostic laboratories. K. H. Chan, K. K. W. To, P. T. W. Li, T. L. Wong, R. Zhang, K. K. H. Chik, G. Chan, C. C. Y. Yip, H. L. Chen, I. F. N. Hung, J. F. W. Chan, and K. Y. Yuen Copyright © 2017 K. H. Chan et al. All rights reserved. Global Status of Porcine circovirus Type 2 and Its Associated Diseases in Sub-Saharan Africa Sun, 12 Mar 2017 00:00:00 +0000 Globally, Porcine circovirus type 2 (PCV2) is a recognized viral pathogen of great economic value in pig farming. It is the major cause of ravaging postweaning multisystemic wasting syndrome (PMWS) and many other disease syndromes generally regarded as Porcine circovirus associated diseases (PCVAD) in Europe. PCV2 infections, specifically PMWS, had impacted huge economic loss on swine production at different regions of the world. It has been studied and reported at different parts of the globe including: North and South America, Europe, Asia, Oceania, Middle East, and the Caribbean. However, till date, this virus and its associated diseases have been grossly understudied in sub-Sahara African region and the entire continent at large. Two out of forty-nine, representing just about 4% of countries that make up sub-Sahara Africa presently, have limited records on reported cases and occurrence of the viral pathogen despite the ubiquitous nature of the virus. This review presents an overview of the discovery of Porcine circovirus and its associated diseases in global pig herds and emphasizes the latest trends in PCV2 vaccines and antiviral drugs development and the information gaps that exist on the occurrence of this important viral pathogen in swine herds of sub-Saharan Africa countries. This will serve as wake-up call for immediate and relevant actions by stakeholders in the region. Kayode O. Afolabi, Benson C. Iweriebor, Anthony I. Okoh, and Larry C. Obi Copyright © 2017 Kayode O. Afolabi et al. All rights reserved. Patterns of Human Respiratory Viruses and Lack of MERS-Coronavirus in Patients with Acute Upper Respiratory Tract Infections in Southwestern Province of Saudi Arabia Mon, 27 Feb 2017 09:54:03 +0000 We undertook enhanced surveillance of those presenting with respiratory symptoms at five healthcare centers by testing all symptomatic outpatients between November 2013 and January 2014 (winter time). Nasal swabs were collected from 182 patients and screened for MERS-CoV as well as other respiratory viruses using RT-PCR and multiplex microarray. A total of 75 (41.2%) of these patients had positive viral infection. MERS-CoV was not detected in any of the samples. Human rhinovirus (hRV) was the most detected pathogen (40.9%) followed by non-MERS-CoV human coronaviruses (19.3%), influenza (Flu) viruses (15.9%), and human respiratory syncytial virus (hRSV) (13.6%). Viruses differed markedly depending on age in which hRV, Flu A, and hCoV-OC43 were more prevalent in adults and RSV, hCoV-HKU1, and hCoV-NL63 were mostly restricted to children under the age of 15. Moreover, coinfection was not uncommon in this study, in which 17.3% of the infected patients had dual infections due to several combinations of viruses. Dual infections decreased with age and completely disappeared in people older than 45 years. Our study confirms that MERS-CoV is not common in the southwestern region of Saudi Arabia and shows high diversity and prevalence of other common respiratory viruses. This study also highlights the importance and contribution of enhanced surveillance systems for better infection control. Ahmed A. Abdulhaq, Vinod Kumar Basode, Anwar M. Hashem, Ahmed S. Alshrari, Nassrin A. Badroon, Ahmed M. Hassan, Tagreed L. Alsubhi, Yahia Solan, Saleh Ejeeli, and Esam I. Azhar Copyright © 2017 Ahmed A. Abdulhaq et al. All rights reserved. Herpes Simplex Virus Latency: The DNA Repair-Centered Pathway Wed, 01 Feb 2017 00:00:00 +0000 Like all herpesviruses, herpes simplex virus 1 (HSV1) is able to produce lytic or latent infections depending on the host cell type. Lytic infections occur in a broad range of cells while latency is highly specific for neurons. Although latency suggests itself as an attractive target for novel anti-HSV1 therapies, progress in their development has been slowed due in part to a lack of agreement about the basic biochemical mechanisms involved. Among the possibilities being considered is a pathway in which DNA repair mechanisms play a central role. Repair is suggested to be involved in both HSV1 entry into latency and reactivation from it. Here I describe the basic features of the DNA repair-centered pathway and discuss some of the experimental evidence supporting it. The pathway is particularly attractive because it is able to account for important features of the latent response, including the specificity for neurons, the specificity for neurons of the peripheral compared to the central nervous system, the high rate of genetic recombination in HSV1-infected cells, and the genetic identity of infecting and reactivated virus. Jay C. Brown Copyright © 2017 Jay C. Brown. All rights reserved. Incidence and Molecular Characterization of Hepatitis E Virus from Swine in Eastern Cape, South Africa Mon, 16 Jan 2017 00:00:00 +0000 Hepatitis E virus-mediated infection is a serious public health concern in economically developing nations of the world. Globally, four major genotypes of HEV have been documented. Hepatitis E has been suggested to be zoonotic owing to the increase of evidence through various studies. Thus far, this paper reports on prevalence of hepatitis E virus among swine herd in selected communal and commercial farms in the Eastern Cape Province of South Africa. A total of 160 faecal samples were collected from swine herds in Amathole and Chris Hani District Municipalities of Eastern Cape Province for the presence of HEV. Of the 160 faecal samples screened, only seven were positive (4.4%) for HEV. The nucleotide sequences analyses revealed the isolates as sharing 82% to 99% identities with other strains (KX896664, KX896665, KX896666, KX896667, KX896668, KX896669, and KX896670) from different regions of the world. We conclude that HEV is present among swine in the Eastern Cape Province, albeit in low incidence, and this does have public health implications. There is a need for maintenance of high hygienic standards in order to prevent human infections through swine faecal materials and appropriate cooking of pork is highly advised. Olusesan Adeyemi Adelabu, Benson Chuks Iweriebor, U. U. Nwodo, Larry Chikwelu Obi, and Anthony Ifeanyi Okoh Copyright © 2017 Olusesan Adeyemi Adelabu et al. All rights reserved. Molecular Detection of Torque Teno Sus Virus and Coinfection with African Swine Fever Virus in Blood Samples of Pigs from Some Slaughterhouses in Nigeria Wed, 19 Oct 2016 09:31:47 +0000 Torque teno sus virus 1 (TTSuV1a/TTSuV1b) infection is present in pig herds worldwide. This study investigated the prevalence of TTSuV1a/TTSuV1b infections in domestic pigs from some slaughterhouses in Nigeria as well as coinfection with African swine fever virus (ASFV) and described the phylogeny in relation to global strains. One hundred and eighty-one (181) blood samples from four slaughterhouses were used for the study and viral nucleic acid detection was carried out by PCR. Comparative sequence analysis was carried out to infer phylogeny. The overall prevalence of TTSuV1a/b was 17.7%. Prevalence of individual genotypes was 10.5% and 7.2% for TTSuV1a and TTSuV1b, respectively. Coinfection of ASFV/TTSuV1a/b was 7.7% while that of TTSuV1a and TTSuV1b was 1.7%. ASFV alone was detected in 11.91% of the total samples. The Nigerian TTSuV1a and TTSuV1b shared a sequence identity of 91–100% and 95–100%, respectively, among each other. The ASFV sequences were 100% identical to members of genotype 1. This is the first report on the presence of TTSuV1a/b in domestic pigs in Nigeria and coinfection with ASFV. Although the prevalence of TTSuV1a/b in Nigeria was low, we recommend further studies to establish the trend and possible role in the pathogenesis of ASFV. Pam D. Luka, Joseph Erume, Bitrus Yakubu, Olajide A. Owolodun, David Shamaki, and Frank N. Mwiine Copyright © 2016 Pam D. Luka et al. All rights reserved. Immunogenicity of RSV F DNA Vaccine in BALB/c Mice Mon, 05 Sep 2016 13:16:41 +0000 Respiratory syncytial virus (RSV) causes severe acute lower respiratory tract disease leading to numerous hospitalizations and deaths among the infant and elderly populations worldwide. There is no vaccine or a less effective drug available against RSV infections. Natural RSV infection stimulates the Th1 immune response and activates the production of neutralizing antibodies, while earlier vaccine trials that used UV-inactivated RSV exacerbated the disease due to the activation of the allergic Th2 response. With a focus on Th1 immunity, we developed a DNA vaccine containing the native RSV fusion (RSV F) protein and studied its immune response in BALB/c mice. High levels of RSV specific antibodies were induced during subsequent immunizations. The serum antibodies were able to neutralize RSV in vitro. The RSV inhibition by sera was also shown by immunofluorescence analyses. Antibody response of the RSV F DNA vaccine showed a strong Th1 response. Also, sera from RSV F immunized and RSV infected mice reduced the RSV infection by 50% and 80%, respectively. Our data evidently showed that the RSV F DNA vaccine activated the Th1 biased immune response and led to the production of neutralizing antibodies, which is the desired immune response required for protection from RSV infections. Erdal Eroglu, Ankur Singh, Swapnil Bawage, Pooja M. Tiwari, Komal Vig, Shreekumar R. Pillai, Vida A. Dennis, and Shree R. Singh Copyright © 2016 Erdal Eroglu et al. All rights reserved. Isolation and Metagenomic Identification of Avian Leukosis Virus Associated with Mortality in Broiler Chicken Mon, 15 Aug 2016 07:14:13 +0000 Avian leukosis virus (ALV) belongs to the family Retroviridae and causes considerable economic losses to the poultry industry. Following an outbreak associated with high mortality in a broiler flock in northern part of Malaysia, kidney tissues from affected chickens were submitted for virus isolation and identification in chicken embryonated egg and MDCK cells. Evidence of virus growth was indicated by haemorrhage and embryo mortality in egg culture. While viral growth in cell culture was evidenced by the development of cytopathic effects. The isolated virus was purified by sucrose gradient and identified using negative staining transmission electron microscopy. Further confirmation was achieved through next-generation sequencing and nucleotide sequence homology search. Analysis of the viral sequences using the NCBI BLAST tool revealed 99-100% sequence homology with exogenous ALV viral envelope protein. Phylogenetic analysis based on partial envelope sequences showed the Malaysian isolate clustered with Taiwanese and Japanese ALV strains, which were closer to ALV subgroup J, ALV subgroup E, and recombinant A/E isolates. Based on these findings, ALV was concluded to be associated with the present outbreak. It was recommended that further studies should be conducted on the molecular epidemiology and pathogenicity of the identified virus isolate. Faruku Bande, Siti Suri Arshad, and Abdul Rahman Omar Copyright © 2016 Faruku Bande et al. All rights reserved. Molecular and Phylogenetic Analysis of Bovine Papillomavirus Type 1: First Report in Iraqi Cattle Mon, 20 Jun 2016 07:51:37 +0000 This study aimed to provide the first molecular characterization of bovine papillomavirus type 1 (BPV-1) in Iraq. BPV is a widely spread oncogenic virus in Iraqi cattle and is associated with the formation of both benign and malignant lesions, resulting in notable economic losses in dairy and beef cattle. In the current study, 140 cutaneous papilloma specimens were collected from cattle in central Iraq. These samples were submitted to histopathological examination, PCR, and sequencing analysis. The histopathology revealed that the main lesion type among the specimens was fibropapilloma. BPV-1 DNA was detected in 121 of the samples (86.42%) in Iraqi cattle as the main causative agent for the disease. A partial sequence for the E2, L2 genes, and complete sequence for the E5 gene were deposited in GenBank. Phylogenetic analysis confirmed the presence of BPV-1 and showed that the origin of infection may be imported European cattle. Obtaining a complete E5 gene sequence enabled us to perform structural predictions. This study presents the first report of BPV-1 infection in the Iraqi cattle and contributes to extending the knowledge of the origin of the spread of this disease. The results of this study will aid in the development of appropriate control measures and therapeutic strategies. Mohammed A. Hamad, Ahmed M. Al-Shammari, Shoni M. Odisho, and Nahi Y. Yaseen Copyright © 2016 Mohammed A. Hamad et al. All rights reserved. Vitamin D-Regulated MicroRNAs: Are They Protective Factors against Dengue Virus Infection? Wed, 11 May 2016 13:46:26 +0000 Over the last few years, an increasing body of evidence has highlighted the critical participation of vitamin D in the regulation of proinflammatory responses and protection against many infectious pathogens, including viruses. The activity of vitamin D is associated with microRNAs, which are fine tuners of immune activation pathways and provide novel mechanisms to avoid the damage that arises from excessive inflammatory responses. Severe symptoms of an ongoing dengue virus infection and disease are strongly related to highly altered production of proinflammatory mediators, suggesting impairment in homeostatic mechanisms that control the host’s immune response. Here, we discuss the possible implications of emerging studies anticipating the biological effects of vitamin D and microRNAs during the inflammatory response, and we attempt to extrapolate these findings to dengue virus infection and to their potential use for disease management strategies. John F. Arboleda and Silvio Urcuqui-Inchima Copyright © 2016 John F. Arboleda and Silvio Urcuqui-Inchima. All rights reserved. A Cross-Study Biomarker Signature of Human Bronchial Epithelial Cells Infected with Respiratory Syncytial Virus Wed, 04 May 2016 13:52:00 +0000 Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract infections in children, elderly, and immunocompromised individuals. Despite of advances in diagnosis and treatment, biomarkers of RSV infection are still unclear. To understand the host response and propose signatures of RSV infection, previous studies evaluated the transcriptional profile of the human bronchial epithelial cell line—BEAS-2B—infected with different strains of this virus. However, the evolution of statistical methods and functional analysis together with the large amount of expression data provide opportunities to uncover novel biomarkers of inflammation and infections. In view of those facts publicly available microarray datasets from RSV-infected BEAS-2B cells were analyzed with linear model-based statistics and the platform for functional analysis InnateDB. The results from those analyses argue for the reevaluation of previously reported transcription patterns and biological pathways in BEAS-2B cell lines infected with RSV. Importantly, this study revealed a biosignature constituted by genes such as ABCC4, ARMC8, BCLAF1, EZH1, FAM118A, FAM208B, FUS, HSPH1, KAZN, MAP3K2, N6AMT1, PRMT2, S100PBP, SERPINA1, TLK2, ZNF322, and ZNF337 which should be considered in the development of new molecular diagnosis tools. Luiz Gustavo Gardinassi Copyright © 2016 Luiz Gustavo Gardinassi. All rights reserved. Direct Detection and Identification of Enteroviruses from Faeces of Healthy Nigerian Children Using a Cell-Culture Independent RT-Seminested PCR Assay Sun, 20 Mar 2016 06:23:44 +0000 Recently, a cell-culture independent protocol for detection of enteroviruses from clinical specimen was recommended by the WHO for surveillance alongside the previously established protocols. Here, we investigated whether this new protocol will show the same enterovirus diversity landscape as the established cell-culture dependent protocols. Faecal samples were collected from sixty apparently healthy children in Ibadan, Nigeria. Samples were resuspended in phosphate buffered saline, RNA was extracted, and the VP1 gene was amplified using WHO recommended RT-snPCR protocol. Amplicons were sequenced and sequences subjected to phylogenetic analysis. Fifteen (25%) of the 60 samples yielded the expected band size. Of the 15 amplicons sequenced, 12 were exploitable. The remaining 3 had electropherograms with multiple peaks and were unexploitable. Eleven of the 12 exploitable sequences were identified as Coxsackievirus A1 (CVA1), CVA3, CVA4, CVA8, CVA20, echovirus 32 (E32), enterovirus 71 (EV71), EVB80, and EVC99. Subsequently, the last exploitable sequence was identified as enterobacteriophage baseplate gene by nucleotide BLAST. The results of this study document the first description of molecular sequence data on CVA1, CVA8, and E32 strains present in Nigeria. The result further showed that species A enteroviruses were more commonly detected in the region when cell-culture bias is bypassed. Temitope Oluwasegun Cephas Faleye, Moses Olubusuyi Adewumi, Bamidele Atinuke Coker, Felix Yasha Nudamajo, and Johnson Adekunle Adeniji Copyright © 2016 Temitope Oluwasegun Cephas Faleye et al. All rights reserved. Serological Survey of Hantavirus in Inhabitants from Tropical and Subtropical Areas of Brazil Thu, 10 Mar 2016 10:08:40 +0000 Brazil has reported more than 1,600 cases of hantavirus cardiopulmonary syndrome (HPS) since 1993, with a 39% rate of reported fatalities. Using a recombinant nucleocapsid protein of Araraquara virus, we performed ELISA to detect IgG antibodies against hantavirus in human sera. The aim of this study was to analyze hantavirus antibody levels in inhabitants from a tropical area (Amazon region) in Rondônia state and a subtropical (Atlantic Rain Forest) region in São Paulo state, Brazil. A total of 1,310 serum samples were obtained between 2003 and 2008 and tested by IgG-ELISA, and 82 samples (6.2%), of which 62 were from the tropical area (5.8%) and 20 from the subtropical area (8.3%), tested positive. Higher levels of hantavirus antibody were observed in inhabitants of the populous subtropical areas compared with those from the tropical areas in Brazil. Felipe Alves Morais, Alexandre Pereira, Aparecida Santo Pietro Pereira, Marcos Lazaro Moreli, Luís Marcelo Aranha Camargo, Marcello Schiavo Nardi, Cristina Farah Tófoli, Jansen Araujo, Lilia Mara Dutra, Tatiana Lopes Ometto, Renata Hurtado, Fábio Carmona de Jesus Maués, Tiene Zingano Hinke, Sati Jaber Mahmud, Monica Correia Lima, Luiz Tadeu Moraes Figueiredo, and Edison Luiz Durigon Copyright © 2016 Felipe Alves Morais et al. All rights reserved. Cumulative Impact of HIV and Multiple Concurrent Human Papillomavirus Infections on the Risk of Cervical Dysplasia Mon, 22 Feb 2016 13:28:22 +0000 Infection with HIV is known to increase the risk of cervical cancer. In addition, evidence suggests that concurrent infection with multiple human papillomavirus (HPV) genotypes increases the risk of cervical dysplasia more than infection with a single HPV genotype. However, the impact of the combination of HIV coinfection and presence of multiple concurrent HPV infections on the risk of cervical dysplasia is uncertain. We compared the results of HPV testing and Pap smears between HIV-infected and HIV-uninfected young women to assess the cumulative impact of these two conditions. We found that both HIV and the presence of multiple concurrent HPV infections are associated with increased risk of associated Pap smear abnormality and that the impact of these two risk factors may be additive. David H. Adler, Melissa Wallace, Thola Bennie, Beau Abar, Tracy L. Meiring, Anna-Lise Williamson, and Linda-Gail Bekker Copyright © 2016 David H. Adler et al. All rights reserved. Adapted Lethality: What We Can Learn from Guinea Pig-Adapted Ebola Virus Infection Model Thu, 18 Feb 2016 15:46:32 +0000 Establishment of small animal models of Ebola virus (EBOV) infection is important both for the study of genetic determinants involved in the complex pathology of EBOV disease and for the preliminary screening of antivirals, production of therapeutic heterologic immunoglobulins, and experimental vaccine development. Since the wild-type EBOV is avirulent in rodents, the adaptation series of passages in these animals are required for the virulence/lethality to emerge in these models. Here, we provide an overview of our several adaptation series in guinea pigs, which resulted in the establishment of guinea pig-adapted EBOV (GPA-EBOV) variants different in their characteristics, while uniformly lethal for the infected animals, and compare the virologic, genetic, pathomorphologic, and immunologic findings with those obtained in the adaptation experiments of the other research groups. S. V. Cheresiz, E. A. Semenova, and A. A. Chepurnov Copyright © 2016 S. V. Cheresiz et al. All rights reserved. Pathogenesis and Diagnostic Approaches of Avian Infectious Bronchitis Wed, 03 Feb 2016 14:15:59 +0000 Infectious bronchitis (IB) is one of the major economically important poultry diseases distributed worldwide. It is caused by infectious bronchitis virus (IBV) and affects both galliform and nongalliform birds. Its economic impact includes decreased egg production and poor egg quality in layers, stunted growth, poor carcass weight, and mortality in broiler chickens. Although primarily affecting the respiratory tract, IBV demonstrates a wide range of tissues tropism, including the renal and reproductive systems. Thus, disease outcome may be influenced by the organ or tissue involved as well as pathotypes or strain of the infecting virus. Knowledge on the epidemiology of the prevalent IBV strains in a particular region is therefore important to guide control and preventions. Meanwhile previous diagnostic methods such as serology and virus isolations are less sensitive and time consuming, respectively; current methods, such as reverse transcription polymerase chain reaction (RT-PCR), Restriction Fragment Length Polymorphism (RFLP), and sequencing, offer highly sensitive, rapid, and accurate diagnostic results, thus enabling the genotyping of new viral strains within the shortest possible time. This review discusses aspects on pathogenesis and diagnostic methods for IBV infection. Faruku Bande, Siti Suri Arshad, Abdul Rahman Omar, Mohd Hair Bejo, Muhammad Salisu Abubakar, and Yusuf Abba Copyright © 2016 Faruku Bande et al. All rights reserved. Serological Investigation of Akabane Virus Infection in Cattle and Sheep in Nigeria Tue, 26 Jan 2016 13:58:15 +0000 Akabane virus (AKAV) is recognized as an important pathogen that causes abortions and congenital malformations in ruminants. However, it has not received adequate attention in Nigeria. Therefore, in investigating this disease, serum samples from 184 (abattoir and farm) head of cattle and 184 intensively reared sheep from two states in southwest Nigeria were screened for antibodies against AKAV using enzyme-linked immunosorbent assay. An overall seropositivity of 70.1% (129/184) was obtained with antibodies being detectable in 73.8% of abattoir (trade) cattle and 40.0% in farm cattle, while 4.3% (8/184) seropositivity was observed in sheep. All the age groups of cattle tested had seropositive animals, 0-1 year (1/7, 14.3%), 2-3 years (17/34, 50.0%), 4-5 years (92/121, 76.0%), and >5 years (19/22, 86.4%), while in sheep only the age groups of 2-3 and 4-5 years showed seropositivity of 4.1% (4/97) and 8.2% (4/49), respectively. The detection of antibody-positive animals among unvaccinated cattle and sheep provides evidence of AKAV infection in Nigeria. These findings call for continuous monitoring of the disease among ruminants in order to ascertain the actual burden and increase awareness of the disease. This will facilitate early detection and aid the development of appropriate control measures against the disease in Nigeria. Daniel Oladimeji Oluwayelu, Comfort Oluladun Aiki-Raji, Emmanuel Chibuzor Umeh, Samat Odunayo Mustapha, and Adebowale Idris Adebiyi Copyright © 2016 Daniel Oladimeji Oluwayelu et al. All rights reserved. Exosomes in Human Immunodeficiency Virus Type I Pathogenesis: Threat or Opportunity? Mon, 04 Jan 2016 13:04:15 +0000 Nanometre-sized vesicles, also known as exosomes, are derived from endosomes of diverse cell types and present in multiple biological fluids. Depending on their cellular origins, the membrane-bound exosomes packed a variety of functional proteins and RNA species. These microvesicles are secreted into the extracellular space to facilitate intercellular communication. Collective findings demonstrated that exosomes from HIV-infected subjects share many commonalities with Human Immunodeficiency Virus Type I (HIV-1) particles in terms of proteomics and lipid profiles. These observations postulated that HIV-resembled exosomes may contribute to HIV pathogenesis. Interestingly, recent reports illustrated that exosomes from body fluids could inhibit HIV infection, which then bring up a new paradigm for HIV/AIDS therapy. Accumulative findings suggested that the cellular origin of exosomes may define their effects towards HIV-1. This review summarizes the two distinctive roles of exosomes in regulating HIV pathogenesis. We also highlighted several additional factors that govern the exosomal functions. Deeper understanding on how exosomes promote or abate HIV infection can significantly contribute to the development of new and potent antiviral therapeutic strategy and vaccine designs. Sin-Yeang Teow, Alif Che Nordin, Syed A. Ali, and Alan Soo-Beng Khoo Copyright © 2016 Sin-Yeang Teow et al. All rights reserved. Expression of Factor X in BHK-21 Cells Promotes Low Pathogenic Influenza Viruses Replication Thu, 31 Dec 2015 14:30:09 +0000 A cDNA clone for factor 10 (FX) isolated from chicken embryo inserted into the mammalian cell expression vector pCDNA3.1 was transfected into the baby hamster kidney (BHK-21) cell line. The generated BHK-21 cells with inducible expression of FX were used to investigate the efficacy of the serine transmembrane protease to proteolytic activation of influenza virus hemagglutinin (HA) with monobasic cleavage site. Data showed that the BHK-21/FX stably expressed FX after ten serial passages. The cells could proteolytically cleave the HA of low pathogenic avian influenza virus at multiplicity of infection 0.01. Growth kinetics of the virus on BHK-21/FX, BHK-21, and MDCK cells were evaluated by titrations of virus particles in each culture supernatant. Efficient multicycle viral replication was markedly detected in the cell at subsequent passages. Virus titration demonstrated that BHK-21/FX cell supported high-titer growth of the virus in which the viral titer is comparable to the virus grown in BHK-21 or MDCK cells with TPCK-trypsin. The results indicate potential application for the BHK-21/FX in influenza virus replication procedure and related studies. Shahla Shahsavandi, Mohammad Majid Ebrahimi, Shahin Masoudi, and Hasan Izadi Copyright © 2015 Shahla Shahsavandi et al. All rights reserved. Antiviral Activity of Resveratrol against Human and Animal Viruses Sun, 29 Nov 2015 11:57:28 +0000 Resveratrol is a potent polyphenolic compound that is being extensively studied in the amelioration of viral infections both in vitro and in vivo. Its antioxidant effect is mainly elicited through inhibition of important gene pathways like the NF-κβ pathway, while its antiviral effects are associated with inhibitions of viral replication, protein synthesis, gene expression, and nucleic acid synthesis. Although the beneficial roles of resveratrol in several viral diseases have been well documented, a few adverse effects have been reported as well. This review highlights the antiviral mechanisms of resveratrol in human and animal viral infections and how some of these effects are associated with the antioxidant properties of the compound. Yusuf Abba, Hasliza Hassim, Hazilawati Hamzah, and Mohamed Mustapha Noordin Copyright © 2015 Yusuf Abba et al. All rights reserved. Clinical Symptoms of Human Rotavirus Infection Observed in Children in Sokoto, Nigeria Wed, 25 Nov 2015 06:26:58 +0000 Rotavirus has been identified among the most important causes of infantile diarrhoea, especially in developing countries. The present study was undertaken to determine the occurrence and clinical symptoms of human rotavirus disease among children presenting with varying degree of diarrhoea in selected urban hospitals in Sokoto metropolis, Nigeria. Diarrhoea samples were collected from 200 diarrheic children younger than 5 years of age and tested using a commercially available DAKO Rotavirus ELISA kit which detects the presence of human group A rotaviruses. A questionnaire, based on WHO generic protocol, was completed for each child to generate the primary data. Of the total number of samples collected, 51 were found to be positive for human group A rotavirus indicating 25.5% prevalence of the disease in Sokoto state. The symptoms associated with the disease were analyzed and discussed. B. R. Alkali, A. I. Daneji, A. A. Magaji, and L. S. Bilbis Copyright © 2015 B. R. Alkali et al. All rights reserved. Evaluating Andrographolide as a Potent Inhibitor of NS3-4A Protease and Its Drug-Resistant Mutants Using In Silico Approaches Mon, 26 Oct 2015 08:31:24 +0000 Current combination therapy of PEG-INF and ribavirin against the Hepatitis C Virus (HCV) genotype-1 infections is ineffective in maintaining sustained viral response in 50% of the infection cases. New compounds in the form of protease inhibitors can complement the combination therapy. Asunaprevir is new to the drug regiment as the NS3-4A protease inhibitor, but it is susceptible to two mutations, namely, R155K and D168A in the protein. Thus, in our study, we sought to evaluate Andrographolide, a labdane-diterpenoid from the Andrographis paniculata plant as an effective compound for inhibiting the NS3-4A protease as well as its concomitant drug-resistant mutants by using molecular docking and dynamic simulations. Our study shows that Andrographolide has best docking scores of −15.0862, −15.2322, and −13.9072 compared to those of Asunaprevir −3.7159, −2.6431, and −5.4149 with wild-type R155K and D168A mutants, respectively. Also, as shown in the MD simulations, the compound was good in binding the target proteins and maintains strong bonds causing very less to negligible perturbation in the protein backbone structures. Our results validate the susceptibility of Asunaprevir to protein variants as seen from our docking studies and trajectory period analysis. Therefore, from our study, we hope to add one more option in the drug regiment to tackle drug resistance in HCV infections. Vivek Chandramohan, Anubhav Kaphle, Mamatha Chekuri, Sindhu Gangarudraiah, and Gowrishankar Bychapur Siddaiah Copyright © 2015 Vivek Chandramohan et al. All rights reserved. Measles Virus: Identification in the M Protein Primary Sequence of a Potential Molecular Marker for Subacute Sclerosing Panencephalitis Mon, 26 Oct 2015 07:16:06 +0000 Subacute Sclerosing Panencephalitis (SSPE), a rare lethal disease of children and young adults due to persistence of measles virus (MeV) in the brain, is caused by wild type (wt) MeV. Why MeV vaccine strains never cause SSPE is completely unknown. Hypothesizing that this phenotypic difference could potentially be represented by a molecular marker, we compared glycoprotein and matrix (M) genes from SSPE cases with those from the Moraten vaccine strain, searching for differential structural motifs. We observed that all known SSPE viruses have residues P64, E89, and A209 (PEA) in their M proteins whereas the equivalent residues for vaccine strains are either S64, K89, and T209 (SKT) as in Moraten or PKT. Through the construction of MeV recombinants, we have obtained evidence that the wt MeV-M protein PEA motif, in particular A209, is linked to increased viral spread. Importantly, for the 10 wt genotypes (of 23) that have had their M proteins sequenced, 9 have the PEA motif, the exception being B3, which has PET. Interestingly, cases of SSPE caused by genotype B3 have yet to be reported. In conclusion, our results strongly suggest that the PEA motif is a molecular marker for wt MeV at risk to cause SSPE. Hasan Kweder, Michelle Ainouze, Joanna Brunel, Denis Gerlier, Evelyne Manet, and Robin Buckland Copyright © 2015 Hasan Kweder et al. All rights reserved. Seroprevalence of Herpes Simplex Virus Infection in HIV Coinfected Individuals in Eastern India with Risk Factor Analysis Mon, 19 Oct 2015 13:01:28 +0000 Herpes simplex virus type 2 (HSV-2) is the cause of most genital herpes while HSV-1 is responsible for orolabial and facial lesions. In immunocompromised individuals, like HIV patients, impaired immunity leads to more frequent symptomatic and asymptomatic HSV infection. Fifty-two blood samples from HIV patients with clinically diagnosed HSV infection were taken as cases, while 45 blood samples each from HIV-infected (HIV control) and noninfected patients without any herpetic lesion (non-HIV control) were taken as control. Serum was tested for IgM and IgG antibodies of both HSV-1 and HSV-2 by ELISA. The seroprevalence was compared among the three groups of study population, considering the demographic and socioeconomic parameters. The HSV-2 IgM was significantly higher () in the HIV patient group (34.6%) than the HIV control (2.2%) and non-HIV control (2.2%) groups, whereas HSV-2 IgG seroprevalence was higher in both HIV patient (61.5%) and HIV control (57.8%) groups than the non-HIV control group (17.8%). The prevalence of HSV-2 was significantly higher in persons with multiple partners and in the reproductive age group. The overall seroprevalence of HSV-1 IgM was too low (<5%), whereas it was too high (about 90%) with HSV-1 IgG in all three study groups. Soumyabrata Nag, Soma Sarkar, Debprasad Chattopadhyay, Sanjoy Bhattacharya, Rahul Biswas, and Manideepa SenGupta Copyright © 2015 Soumyabrata Nag et al. All rights reserved. Comparison of β-Propiolactone and Formalin Inactivation on Antigenicity and Immune Response of West Nile Virus Thu, 27 Aug 2015 14:18:28 +0000 West Nile Virus (WNV) is a pathogenic arbovirus that belongs to genus Flavivirus under family Flaviviridae. Till now there are no approved vaccines against WNV for human use. In this study, the effect of two alkylating agents, formaldehyde and β-PL, generally used for inactivated vaccine preparation, was assessed on the basis of antigenic and immunogenic potential of the inactivated WNV. Lineage 5 WNV isolates were inactivated by both formalin and β-PL treatments. Inactivation was confirmed by repeated passage in BHK-21 cell line and infant mice. Viruses inactivated by both the treatments showed higher antigenicity. Immune response in mice model showed serum anti-WNV antibody titre was moderately higher in formalin inactivated antigen compared to β-PL inactivated antigen. However, no significant differences were observed in neutralization antibody titre. In conclusion, we can state that both formaldehyde and β-PL inactivation processes were found to be equally efficient for inactivation of WNV. However, they need to be compared with other inactivating agents along with study on cell mediated immune response. Pritom Chowdhury, Rashmee Topno, Siraj A. Khan, and Jagadish Mahanta Copyright © 2015 Pritom Chowdhury et al. All rights reserved. Investigation of Stilbenoids as Potential Therapeutic Agents for Rotavirus Gastroenteritis Wed, 26 Aug 2015 06:54:19 +0000 Rotavirus (RV) infections cause severe diarrhea in infants and young children worldwide. Vaccines are available but cost prohibitive for many countries and only reduce severe symptoms. Vaccinated infants continue to shed infectious particles, and studies show decreased efficacy of the RV vaccines in tropical and subtropical countries where they are needed most. Continuing surveillance for new RV strains, assessment of vaccine efficacy, and development of cost effective antiviral drugs remain an important aspect of RV studies. This study was to determine the efficacy of antioxidant and anti-inflammatory stilbenoids to inhibit RV replication. Peanut (A. hypogaea) hairy root cultures were induced to produce stilbenoids, which were purified by high performance countercurrent chromatography (HPCCC) and analyzed by HPLC. HT29.f8 cells were infected with RV in the presence stilbenoids. Cell viability counts showed no cytotoxic effects on HT29.f8 cells. Viral infectivity titers were calculated and comparatively assessed to determine the effects of stilbenoid treatments. Two stilbenoids, trans-arachidin-1 and trans-arachidin-3, show a significant decrease in RV infectivity titers. Western blot analyses performed on the infected cell lysates complemented the infectivity titrations and indicated a significant decrease in viral replication. These studies show the therapeutic potential of the stilbenoids against RV replication. Judith M. Ball, Fabricio Medina-Bolivar, Katelyn Defrates, Emily Hambleton, Megan E. Hurlburt, Lingling Fang, Tianhong Yang, Luis Nopo-Olazabal, Richard L. Atwill, Pooja Ghai, and Rebecca D. Parr Copyright © 2015 Judith M. Ball et al. All rights reserved. Farming of Plant-Based Veterinary Vaccines and Their Applications for Disease Prevention in Animals Thu, 13 Aug 2015 12:14:49 +0000 Plants have been studied for the production of pharmaceutical compounds for more than two decades now. Ever since the plant-made poultry vaccine against Newcastle disease virus made a breakthrough and went all the way to obtain regulatory approval, research to use plants for expression and delivery of vaccine proteins for animals was intensified. Indeed, in view of the high production costs of veterinary vaccines, plants represent attractive biofactories and offer many promising advantages in the production of recombinant vaccine proteins. Furthermore, the possibility of conducting immunogenicity and challenge studies in target animals has greatly exaggerated the progress. Although there are no edible plant-produced animal vaccines in the market, plant-based vaccine technology has great potentials. In this review, development, uses, and advantages of plant-based recombinant protein production in various expression platforms are discussed. In addition, examples of plant-based veterinary vaccines showing strong indication in terms of efficacy in animal disease prevention are also described. Pit Sze Liew and Mohd Hair-Bejo Copyright © 2015 Pit Sze Liew and Mohd Hair-Bejo. All rights reserved. Viral Etiology of Chronic Obstructive Pulmonary Disease Exacerbations during the A/H1N1pdm09 Pandemic and Postpandemic Period Thu, 07 May 2015 07:02:38 +0000 Viral infections are one of the main causes of acute exacerbations of chronic obstructive pulmonary disease (AE-COPD). Emergence of A/H1N1pdm influenza virus in the 2009 pandemic changed the viral etiology of exacerbations that were reported before the pandemic. The aim of this study was to describe the etiology of respiratory viruses in 195 Spanish patients affected by AE-COPD from the pandemic until the 2011-12 influenza epidemic. During the study period (2009–2012), respiratory viruses were identified in 48.7% of samples, and the proportion of viral detections in AE-COPD was higher in patients aged 30–64 years than ≥65 years. Influenza A viruses were the pathogens most often detected during the pandemic and the following two influenza epidemics in contradistinction to human rhino/enteroviruses that were the main viruses causing AE-COPD before the pandemic. The probability of influenza virus detection was 2.78-fold higher in patients who are 30–64 years old than those ≥65. Most respiratory samples were obtained during the pandemic, but the influenza detection rate was higher during the 2011-12 epidemic. There is a need for more accurate AE-COPD diagnosis, emphasizing the role of respiratory viruses. Furthermore, diagnosis requires increased attention to patient age and the characteristics of each influenza epidemic. Ivan Sanz, Sonia Tamames, Silvia Rojo, Mar Justel, José Eugenio Lozano, Carlos Disdier, Tomás Vega, and Raúl Ortiz de Lejarazu Copyright © 2015 Ivan Sanz et al. All rights reserved.