ABSTRACT
Effect of Aluminium – Magnesium Silicate (AMS) on Avian Influenza Virus was tested in vitro by incubating an isolate of a highly pathogenic Avian Influenza Virus (AIV) involved in an outbreak of the disease in Nigeria, with a synthetic AMS. The isolate was a confirmed Influenza virus type A, subtype H5N1. Equal amount of the Virus sample and of the AMS, on volume to weight (v/w) basis, were incubated at room temperature for one hour and then centrifuged at 3000 revolutions per minute. The supernatant was tested for viral haemagglutination (HA) titre and for Mean Death Time (MDT) and Embryo Mortality rate (EMR) in embryonated chicken eggs. Incubating samples of the AIV isolate with the synthetic AMS, reduced volumes of the samples at mean rate of 24.3%. It also reduced the viral (HA) titre from a mean of 73±32.72 to 1.4 ±0.43 (P<0.05) and EMR from 100% to 65%. MDT of chicken embryos innocuated with the AIV increased from 76 ±4.38 hours to 130 ±17.27 hours (P< 0.05). When three portions of a sample of the AIV were incubated with the AMS once, twice and thrice respectively,MDT of chick embryos innoculated with the isolate increased from 64 hours to 104 hours in the portion incubated once. In the two portions where the incubation was repeated EMR reduced from 100% to zero.
Chapter One
1.0 INTRODUCTION:
1.1 Avian influenza
Viruses forming the Orthomyxoviridae are enveloped RNA viruses with single stranded genome of negative sense (i.e. the virus RNA is complementary to the messenger RNA) which are divided into eight segments. Proteins are associated with the RNA genome to form the nucleoprotein-RNA-polymerase complex (Jordan, 1999).
The matrix protein surrounding the genome complex is enveloped in a lipid membrane which is covered by two different surface projection proteins with which haemagglutinin and neuraminidase activities are associated, separately. The gene for each of the structural proteins, or its precursor protein, is located on a separate genome segment. This segmentation is an important property of the influenza viruses because, it allows reassortment to occur if two of the viruses infect and replicate in the same cell at the same time (Jordan, 1999).
By negative contrast electron microscopy, Orthomyxoviruses appear as roughly spherical or filamentous particles, 80 – 120nm in diameter on cross-section (Jordan, 1999).
At present, the Orthomyxoviridae family consists of one genus, formed from influenza type A and influenza type B viruses. Influenza type C viruses represent probable, a second genus. To date, only influenza A viruses have been isolated from birds. The Viruses are typed into A, B or C on the basis of the nucleocapsid or matrix antigen which they pocess (Jordan,1999).
Influenza viruses are divided into types A, B, and C based on antigenic differences of nucleocapsid and matrix proteins. The most important is subtype A. Influenza A viruses infect avian species, humans, and many other mammals (Jordan, 1999).
Influenza B viruses affect primarily humans. Influenza C viruses affect humans and swine (Webster, 1999). Serologically, influenza viruses are categorized into 16
Haemagglutinin (H1–H16) and 9 Neuraminidase (N1 – N9) subtypes (Webster, 1999).
The first official report of highly pathogenic avian influenza (HPAI) subtype (H5N1) in Africa was made in February 2006, from outbreak of the disease in a commercial poultry farm in Jaji, Kaduna state, Nigeria (Joannis et al., 2006). The disease on the farm, involved a breeder flock, a laying flock, ostriches, and ducks . The farm attendants also reported outbreaks of similar magnitude in their local chicken flocks. Over forty thousand birds of different species either died or were culled as a result of that outbreak (Joannis et al., 2006).
Surveillance activities in 2008 revealed presence of a new genotype of the virus from a state in Northeastern part of Nigeria that never recorded HPAI outbreak. The genotype was found to be almost identical to A/Cygnusolat/CzechRep/10732/2007 isolated in Europe (Monne et al., 2008).
All Nigerian Avian Influenza Virus isolates were closely related to Influenza viruses that have been circulating in birds in Europe, Russia, and Middle East since 2005 (Monne et al., 2008).According to the unified nomenclature system for highly pathogenic avian influenza (H5N1), these isolates belong to clade 2.2(Monne et al., 2008).
In Nigeria, it is often not easy to keep off feral or other wild birds from commercial farms. So control of AIV is made more difficult. Also, AIV is a zoonotic disease, thus there exist need to research for a possable cure for the disease, to help in controlling spread in birds during outbreaks.
Molecules of Aluminum-Magnesium Silicate (AMS) are reported to have both negative and positive electrical charges (Vanderbilt 1995). Viruses also have charges (Katherine et al., 1985). Thus, it was reseasoned that extracellular viruses may adsorb onto AMS and
so would not infect animal cells.Already, the AMS has been reported to inhibit activities of Newcastle Disease Virus and Egg Drop Syndrome 76 Virus (Ezeibe et al., 20110., Ezeibe, et al., 2011).
2.0 Aims and objectives
-To investigate effect of Synthetic Aluminium – Magnesium Silicate on titre of Avian
Influenza virus.
-To investigate the effect of Synthetic Aluminium – Magnesium Silicate on Embryo
Mortality Rate of chicken eggs inoculated with Avian Influenza Virus.
-To investigate the effect of Synthetic Aluminium – Magnesium Silicate on Mean Death
Time of chicken embryos inoculated with Avian Influenza Virus.
This material content is developed to serve as a GUIDE for students to conduct academic research
EFFECT OF ALUMINIUM MAGNESIUM SILICATE ON AVIAN INFLUENZA VIRUS SUB TYPE H5N1
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