A common approach for developing diagnostic tests for influenza virus detection is the use of mouse or rabbit monoclonal and/or polyclonal antibodies against a target antigen of the virus. However, comparative mapping of the target antigen using antibodies from different animal sources has not been evaluated before. This is important because identification of antigenic determinants of the target antigen in different species plays a central role to ensure the efficiency of a diagnostic test, such as competitive ELISA or immunohistochemistry-based tests. Interest in the matrix 2 ectodomain (M2e) protein of avian influenza virus (AIV) as a candidate for a universal vaccine and also as a marker for detection of virus infection in vaccinated animals (DIVA) is the rationale for the selection of this protein for comparative mapping evaluation. This study aimed to map the epitopes of the M2e protein of avian influenza virus H5N1 using chicken, mouse and rabbit monoclonal or monospecific antibodies. Our findings revealed that rabbit antibodies (rAbs) recognized epitope 6EVETPTRN13 of the M2e, located at the N-terminal of the protein, while mouse (mAb) and chicken antibodies (cAbs) recognized epitope 10PTRNEWECK18, located at the centre region of the protein. The findings highlighted the difference between the M2e antigenic determinants recognized by different species that emphasized the importance of comparative mapping of antibody reactivity from different animals to the same antigen, especially in the case of multi-host infectious agents such as influenza. The findings are of importance for antigenic mapping, as well as diagnostic test and vaccine development.
Fruit phenotypes are often hypothesised to be affected by selection by frugivores. Here, we tested two hypotheses concerning frugivore-fruit interactions from the perspective of fruit colours. We measured the spectral properties of 26 fruits and the associated leaves of plants from 2 islands in New Zealand. Visual observations were also performed to record the birds that fed on the fruits. First, we tested the fruit-foliage hypothesis, where fruit colours are assumed to be evolutionarily constrained by their own leaf colour to maximise colour contrast and fruit conspicuousness. We ran a null model analysis comparing fruit colour contrast using an avian eye model. Second, we tested the frugivore specificity hypothesis, where specific fruit colours are thought to be connected with a specific bird frugivore. We performed a regression on the number of bird visits against the fruit colour in tetrahedral colour space based on an avian eye calculation using Mantel's test. The results show that fruit colours are not constrained by their own leaf colours. There is also no relationship or pattern suggesting a link between a specific fruit colour and specific bird visitors. We suggest that although fruit colour is one of the most highly discussed components, it is not the most important single deciding factor in frugivore fruit selection.
West Nile virus (WNV) is a zoonotic mosquito-borne flavivirus that is harbored and amplified by wild birds via the enzootic transmission cycle. Wide range of hosts are found to be susceptible to WNV infection including mammals, amphibians and reptiles across the world. Several studies have demonstrated that WNV was present in the Malaysian Orang Asli and captive birds. However, no data are available on the WNV prevalence in wild birds found in Malaysia. Therefore this study was conducted to determine the serological and molecular prevalence of WNV in wild birds in selected areas in the West Coast of Peninsular Malaysia. Two types of wild birds were screened, namely migratory and resident birds in order to explore any possibility of WNV transmission from the migratory birds to the resident birds. Thus, a cross-sectional study was conducted at the migratory birds sanctuary located in Kuala Gula, Perak and Kapar, Selangor by catching 163 migratory birds, and 97 resident birds from Kuala Gula and Parit Buntar, Perak at different time between 2016 and 2017 (Total, n = 260). Blood and oropharyngeal swabs were collected for serological and molecular analysis, respectively. Serum were screened for WNV antibodies using a commercial competitive ELISA (c-ELISA) (ID Screen® West Nile Competition Multi-species ELISA, ID VET, Montpellier, France) and cross-reactivity towards Japanese Encephalitis virus (JEV) was also carried out using the JEV-double antigen sandwich (DAS) ELISA. Oropharyngeal swabs were subjected to one-step RT-PCR to detect WNV RNA, in which positive reactions were subsequently sequenced. WNV seropositive rate of 18.71% (29/155) at 95% CI (0.131 to 0.260) and molecular prevalence of 15.2% (16/105) at 95% CI (0.092 to 0.239) were demonstrated in migratory and resident wild birds found in West Coast Malaysia. Phylogenetic analyses of the 16 WNV isolates found in this study revealed that the local strains have 99% similarity to the strains from South Africa and were clustered under lineage 2. Evidence of WNV infection in resident and migratory birds were demonstrated in this study. As a summary, intervention between migratory birds, resident birds and mosquitoes might cause the introduction and maintenance of WNV in Malaysia, however the assumption could be further proven by studying the infection dynamics in the mosquitoes present in the studied areas.
Large, fruit-eating vertebrates have been lost from many of the world's ecosystems. The ecological consequences of this defaunation can be severe, but the evolutionary consequences are nearly unknown because it remains unclear whether frugivores exert strong selection on fruit traits. I assessed the macroevolution of fruit traits in response to variation in the diversity and size of seed-dispersing vertebrates. Across the Indo-Malay Archipelago, many of the same plant lineages have been exposed to very different assemblages of seed-dispersing vertebrates. Phylogenetic analysis of >400 plant species in 41 genera and five families revealed that average fruit size tracks the taxonomic and functional diversity of frugivorous birds and mammals. Fruit size was 40.2-46.5% smaller in the Moluccas and Sulawesi (respectively), with relatively depauperate assemblages of mostly small-bodied animals, than in the Sunda Region (Borneo, Sumatra, and Peninsular Malaysia), with a highly diverse suite of large and small animals. Fruit color, however, was unrelated to vertebrate diversity or to the representation of birds versus mammals in the frugivore assemblage. Overhunting of large animals, nearly ubiquitous in tropical forests, could strongly alter selection pressures on plants, resulting in widespread, although trait-specific, morphologic changes.
Extraintestinal pathogenic Escherichia coli (ExPEC) strains of certain genetic lineages are frequently implicated in a wide range of diseases in humans and birds. ExPEC strains belonging to the phylogenetic lineage/sequence type complex 95 (STC95) are one such prominent lineage that is commonly isolated from extraintestinal infections such as systemic disease in poultry and urinary tract infections (UTIs), neonatal meningitis and sepsis in humans. Several epidemiological studies have indicated that ST95 strains obtained from such infections may share similar virulence genes and other genomic features. However, data on their ability to establish infections in vivo as deduced from the manifestation of similar virulence phenotypes remain elusive. In the present study, 116 STC95 ExPEC isolates comprising 55 human and 61 avian strains, possessing similar virulence gene patterns, were characterized in vitro using adhesion, invasion, biofilm formation and serum bactericidal assays. Overall, STC95 strains from both groups, namely human and birds, were equally capable of adhering to and invading the two mammalian kidney cell lines. Similarly, these strains were able to form strong biofilms in M63 medium. Furthermore, they were equally resistant to the bactericidal activity of human and avian serum. Our cumulative data reinforce the understanding that ST95 strains from poultry present a potential zoonotic risk and therefore need a One Health strategy for a successfull intervention.
Graphene/zinc oxide nanocomposite was synthesised via a facile, green and efficient approach consisted of novel liquid phase exfoliation and solvothermal growth for sensing application. Highly pristine graphene was synthesised through mild sonication treatment of graphite in a mixture of ethanol and water at an optimum ratio. The X-ray diffractometry (XRD) affirmed the hydrothermal growth of pure zinc oxide nanoparticles from zinc nitrate hexahydrate precursor. The as-prepared graphene/zinc oxide (G/ZnO) nanocomposite was characterised comprehensively to evaluate its morphology, crystallinity, composition and purity. All results clearly indicate that zinc oxide particles were homogenously distributed on graphene sheets, without any severe aggregation. The electrochemical performance of graphene/zinc oxide nanocomposite-modified screen-printed carbon electrode (SPCE) was evaluated using cyclic voltammetry (CV) and amperometry analysis. The resulting electrode exhibited excellent electrocatalytic activity towards the reduction of hydrogen peroxide (H2O2) in a linear range of 1-15 mM with a correlation coefficient of 0.9977. The sensitivity of the graphene/zinc oxide nanocomposite-modified hydrogen peroxide sensor was 3.2580 μAmM(-1) with a limit of detection of 7.4357 μM. An electrochemical DNA sensor platform was then fabricated for the detection of Avian Influenza H5 gene based on graphene/zinc oxide nanocomposite. The results obtained from amperometry study indicate that the graphene/zinc oxide nanocomposite-enhanced electrochemical DNA biosensor is significantly more sensitive (P
Reports of bird flu outbreaks occur around the globe, posing a severe threat to the poultry
industry and consumer health. Despite the number of bird flu scares, few studies have
considered the impact on consumers’ chicken consumption intentions in a non-western context,
such as in Malaysia. The aim of this study is to determine the factors that influence consumers’
intentions to take precautions by avoiding the consumption of broiler chicken meat and
products in Malaysia when a bird flu outbreak occurs. A total of 276 online questionnaires were
disseminated using convenience sampling with 109 respondents completing the questionnaires, generating a response rate of 40.8 percent. The study results show that attitudes, subjective norms, perceived behavioral control, perceived credibility of information, and perceived risk have a significant relationship with consumer intentions to take precautions by avoiding the consumption of broiler chicken meat and products. One factor is the exception: attention towards bird flu news. These findings shed some light on understanding the extent of consumer response (i.e. consumption behavior) to future health crises (e.g. bird flu). Moreover, for policy makers, these findings may serve as a guide and a reference to food safety issues in the poultry industry, especially future bird flu scares.
The rapid diagnosis and subtyping of influenza is particularly important in areas where avian influenza (H5N1) is present. The ability to recognise both typical and atypical presentations of influenza is also critical in such settings. A six-month-old male child who visited a H5N1-affected area subsequently died from a severe febrile diarrhoeal illness with minimal respiratory symptoms, and was initially diagnosed with influenza A of an unknown subtype. The final microbiological results showed a highly unusual combination of influenza A (H3N2) and Campylobacter jejuni infection.
A series of plasmids containing the HSP70 gene of Mycobacterium tuberculosis fused to the hemagglutinin (H5) gene of H5N1 avian influenza virus (AIV) (H5-HSP70 (heat shock protein 70) vaccine) or individual H5 gene (H5 vaccine) or HSP70 gene (HSP70 vaccine) were constructed based on the plasmid pcDNA3.1. Expression of H5 gene in Vero cells in vitro and in chickens in vivo was confirmed following their transfection and immunization with H5 or H5-HSP70 vaccines. Controls consisted of HSP70 vaccine, empty plasmid pcDNA3.1 and co-administered H5 and HSP70 vaccines. H5-HSP70 vaccine produced in chicken higher hemagglutination inhibition (HI) antibody titer than H5 vaccine. However, the increase was not statistically significant. We have demonstrated for the first time that the H5 DNA vaccine with fused HSP70 gene may produce an enhanced induction of humoral immune response to AIV in chickens.
Matched MeSH terms: Influenza in Birds/immunology; Influenza in Birds/prevention & control; Influenza in Birds/virology
In this short review, the current knowledge on the botany, ecology, uses, and medicinal properties of the multipurpose Garcinia subelliptica (Fukugi) is updated. As yet, there are no reviews on this indigenous and heritage coastal tree species of the Ryukyu Islands in Japan, which has ethnocultural, ecological, and pharmacological significance. Planted by the Okinawan people some 300 years ago, Fukugi trees serve as windbreaks and accord protection against the destructive typhoons. The species has become a popular ornamental tree, and its bark has been used for dyeing fabrics. It forms part of the food chain for mammals and insects and serves as nesting sites for birds. Endowed with bioactive compounds of benzophenones, xanthones, biflavonoids, and triterpenoids, G. subelliptica possesses anticancer, anti-inflammatory, anti-tyrosinase, trypanocidal, antibacterial, DNA topoisomerase inhibitory, DNA strand scission, choline acetyltransferase enhancing, hypoxia-inducible factor-1 inhibitory, and antiandrogenic activities. Fukugetin and fukugiside are two novel biflavonoids named after the species. The chemical constituents of Fukugi fruits when compared with those of mangosteen yielded interesting contrasts.
Edible bird's nest (EBN) is a popular delicacy in the Asian Pacific region originating from Indonesia, Malaysia, Thailand and Vietnam, which consist of various potential medicine value in Traditional Chinese Medicine (TCM). Thailand is one of the main exporters of EBN. However, the genetic information of EBN, a key part of molecular biology, has yet to be reported in Thailand. It is necessary to explore the genetic information of EBN in Thailand based on a quick and simple method to help protect the rights and interests of consumers. This research aimed to systematically evaluate different methods of extracting EBN DNA to improve the efficiency of the analysis of cytochrome b (Cytb) and NADH dehydrogenase subunit 2 (ND2) gene sequences, the establishment of phylogenetic trees, and the genetic information of EBN in Thailand. Additionally, we aimed to develop a quick and simple method for identifying EBN from different species based on the genetic information and amplification-refractory mutation system PCR (ARMS-PCR). By comparing the four methods [cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), kit and guanidinium isothiocyanate methods] for EBN extraction, we found that the guanidinium isothiocyanate method was the optimal extraction method. Phylogenetic trees generated on the basis of Cytb and ND2 gene analyses showed that 26 samples of house EBN and 4 samples of cave EBN came from Aerodramus fuciphagus and Aerodramus maximus, respectively. In addition, to distinguish different samples from different species of Apodiformes, we designed 4 polymerase chain reaction (PCR) amplification primers based on the ND2 gene sequences of A. fuciphagus and A. maximus. The ARMS-PCR results showed band lengths for A. fuciphagus EBN of 533, 402, and 201 bp, while those for A. maximus EBN were 463, 317, and 201 bp. Collectively, the results showed that ARMS-PCR is a fast and simple method for the genetic identification of EBN based on designing specific original identification primers.
The knowledge of the diversity of haemosporidian parasites is of primary importance as their representatives include agents of bird malaria. We investigated the occurrence of Haemoproteus spp. and Plasmodium spp. in bird populations from a single locality in the State of Selangor, Peninsular Malaysia, and report on the parasite prevalence of the two genera. A combination of methods (molecular and morphological) was used for detecting these parasites. Seventy-nine bird individuals were caught using mist-nets in July and August 2010 at Gombak Field Station of the University of Malaya, Kuala Lumpur. In total, 23 birds were identified as positive for Haemoproteus or Plasmodium infection and one individual was recognized as carrying mixed infection. The total prevalence of haemosporidians in the collected samples was 30.3%. Infections with parasites of the genus Haemoproteus were predominant compared to those of the genus Plasmodium. In total, 10 new cyt b lineages of Haemoproteus spp. and 3 new cyt b lineages of Plasmodium spp. were recorded in this study. From all recorded haemosporidian lineages (16 in total), 3 were known from previous studies - hCOLL2, hYWT2 and pNILSUN1. Two of them are linked with their corresponding morphospecies - Haemoproteus pallidus (COLL2) and Haemoproteus motacillae (YWT2). The morphological analysis in the present study confirmed the results obtained by the PCR method relative to prevalence, with 25.3% total prevalence of Haemoproteus and Plasmodium parasites. The intensities of infection varied between 0.01% and 19%. Most infections were light, with intensities below 0.1%. The present study is the first molecular survey of the protozoan blood parasites of the order Haemosporida recorded in Malaysia.
Since the first H5N1 highly pathogenic avian influenza virus (HPAIV) infection in the region in August 2003, Cambodia, Laos, Malaysia, Myanmar, Indonesia, Thailand and Vietnam have all recorded outbreaks of the disease. The HPAIV continues to occur in some countries in Southeast Asia despite control programmes encompassing surveillance, vaccination and stamping out strategies. A number of strains have been circulating in the region since the first outbreaks in 2003, and although the source of the initial outbreaks in domestic poultry is not known, the continuing propagation of disease in the region is primarily the result of the movement of domestic poultry and poultry products, and people. A comprehensive approach using all the strategies available to break the chain of transmission of the virus in poultry will be needed to achieve lasting disease control.
Matched MeSH terms: Birds; Influenza in Birds/epidemiology*; Influenza in Birds/prevention & control
Habitat degradation and hunting have caused the widespread loss of larger vertebrate species (defaunation) from tropical biodiversity hotspots. However, these defaunation drivers impact vertebrate biodiversity in different ways and, therefore, require different conservation interventions. We conducted landscape-scale camera-trap surveys across six study sites in Southeast Asia to assess how moderate degradation and intensive, indiscriminate hunting differentially impact tropical terrestrial mammals and birds. We found that functional extinction rates were higher in hunted compared to degraded sites. Species found in both sites had lower occupancies in the hunted sites. Canopy closure was the main predictor of occurrence in the degraded sites, while village density primarily influenced occurrence in the hunted sites. Our findings suggest that intensive, indiscriminate hunting may be a more immediate threat than moderate habitat degradation for tropical faunal communities, and that conservation stakeholders should focus as much on overhunting as on habitat conservation to address the defaunation crisis.
We report a case of disseminated histoplasmosis, initially diagnosed from gut nodule colonoscopically, along with evidence of lung disease. Subsequently he developed Addisonian crisis due to adrenal involvement. Lessons were learnt from the importance of detailed history-personal and social/family, as well as psychosocial aspects of illness. Public health measures to reduce bird dropping (and thus possibility of histoplasmosis) are discussed.
Suspicious hybrids of painted storks and milky storks were found in a Malaysian zoo. Blood of these birds was sampled on FTA cards for DNA fingerprinting. Of 44 optimized primers, 6 produced diagnostic markers that could identify hybrids. The markers were based on simple, direct PCR-generated multilocus banding patterns that provided two sets of genetic data, one for each of the two stork species and another for the hybrids. It also revealed that large DNA fragments (3,000 bp) could be amplified from blood collected on FTA cards. When the results of each individual bird's DNA fingerprint were compared with plumage characters, the hybrids were found to express a range of intermediate phenotypic traits of the pure breeds with no dominant plumage characteristic from either parental species.
Virulent Newcastle disease virus (NDV) isolates from new sub-genotypes within genotype VII are rapidly spreading through Asia and the Middle East causing outbreaks of Newcastle disease (ND) characterized by significant illness and mortality in poultry, suggesting the existence of a fifth panzootic. These viruses, which belong to the new sub-genotypes VIIh and VIIi, have epizootic characteristics and do not appear to have originated directly from other genotype VII NDV isolates that are currently circulating elsewhere, but are related to the present and past Indonesian NDV viruses isolated from wild birds since the 80s. Viruses from sub-genotype VIIh were isolated in Indonesia (2009-2010), Malaysia (2011), China (2011), and Cambodia (2011-2012) and are closely related to the Indonesian NDV isolated in 2007, APMV1/Chicken/Karangasem, Indonesia (Bali-01)/2007. Since 2011 and during 2012 highly related NDV isolates from sub-genotype VIIi have been isolated from poultry production facilities and occasionally from pet birds, throughout Indonesia, Pakistan and Israel. In Pakistan, the viruses of sub-genotype VIIi have replaced NDV isolates of genotype XIII, which were commonly isolated in 2009-2011, and they have become the predominant sub-genotype causing ND outbreaks since 2012. In a similar fashion, the numbers of viruses of sub-genotype VIIi isolated in Israel increased in 2012, and isolates from this sub-genotype are now found more frequently than viruses from the previously predominant sub-genotypes VIId and VIIb, from 2009 to 2012. All NDV isolates of sub-genotype VIIi are approximately 99% identical to each other and are more closely related to Indonesian viruses isolated from 1983 through 1990 than to those of genotype VII, still circulating in the region. Similarly, in addition to the Pakistani NDV isolates of the original genotype XIII (now called sub-genotype XIIIa), there is an additional sub-genotype (XIIIb) that was initially detected in India and Iran. This sub-genotype also appears to have as an ancestor a NDV strain from an Indian cockatoo isolated in 1982. These data suggest the existence of a new panzootic composed of viruses of subgenotype VIIi and support our previous findings of co-evolution of multiple virulent NDV genotypes in unknown reservoirs, e.g. as recorded with the virulent NDV identified in Dominican Republic in 2008. The co-evolution of at least three different sub-genotypes reported here and the apparent close relationship of some of those genotypes from ND viruses isolated from wild birds, suggests that identifying wild life reservoirs may help predict new panzootics.
The transmission of highly pathogenic avian influenza H5N1 virus to Southeast Asian countries triggered the first major outbreak and transmission wave in late 2003, accelerating the pandemic threat to the world. Due to the lack of influenza surveillance prior to these outbreaks, the genetic diversity and the transmission pathways of H5N1 viruses from this period remain undefined. To determine the possible source of the wave 1 H5N1 viruses, we recently conducted further sequencing and analysis of samples collected in live-poultry markets from Guangdong, Hunan, and Yunnan in southern China from 2001 to 2004. Phylogenetic analysis of the hemagglutinin and neuraminidase genes of 73 H5N1 isolates from this period revealed a greater genetic diversity in southern China than previously reported. Moreover, results show that eight viruses isolated from Yunnan in 2002 and 2003 were most closely related to the clade 1 virus sublineage from Vietnam, Thailand, and Malaysia, while two viruses from Hunan in 2002 and 2003 were most closely related to viruses from Indonesia (clade 2.1). Further phylogenetic analyses of the six internal genes showed that all 10 of those viruses maintained similar phylogenetic relationships as the surface genes. The 10 progenitor viruses were genotype Z and shared high similarity (>/=99%) with their corresponding descendant viruses in most gene segments. These results suggest a direct transmission link for H5N1 viruses between Yunnan and Vietnam and also between Hunan and Indonesia during 2002 and 2003. Poultry trade may be responsible for virus introduction to Vietnam, while the transmission route from Hunan to Indonesia remains unclear.
Matched MeSH terms: Influenza in Birds/epidemiology; Influenza in Birds/virology*