Application of loop-mediated isothermal amplification assay in the detection of herpesvirus of turkey (FC 126 strain) from chicken samples in Nigeria.

AIM: This study was designed to optimize and apply the use of loop-mediated isothermal amplification (LAMP) as an alternative to conventional polymerase chain reaction (PCR) for the detection of herpesvirus of turkeys (HVT) (FC 126 strain) in vaccinated and non-vaccinated poultry in Nigeria. MATERIALS AND METHODS: HVT positive control (vaccine) was used for optimization of LAMP using six primers that target the HVT070 gene sequence of the virus. These primers can differentiate HVT, a Marek's disease virus (MDV) serotype 3 from MDV serotypes 1 and 2. Samples were collected from clinical cases of Marek's disease (MD) in chickens, processed and subjected to LAMP and PCR. RESULTS: LAMP assay for HVT was optimized. HVT was detected in 60% (3/5) and 100% (5/5) of the samples analyzed by PCR and LAMP, respectively. HVT was detected in the feathers, liver, skin, and spleen with average DNA purity of 3.05-4.52 µg DNA/mg (A260/A280) using LAMP. Conventional PCR detected HVT in two vaccinated and one unvaccinated chicken samples, while LAMP detected HVT in two vaccinated and three unvaccinated corresponding chicken samples. However, LAMP was a faster and simpler technique to carry out than PCR. CONCLUSION: LAMP assay for the detection of HVT was optimized. LAMP and PCR detected HVT in clinical samples collected. LAMP assay can be a very good alternative to PCR for detection of HVT and other viruses. This is the first report of the use of LAMP for the detection of viruses of veterinary importance in Nigeria. LAMP should be optimized as a diagnostic and research tool for investigation of poultry diseases such as MD in Nigeria.

Situation-based survey of avian influenza viruses in possible "bridge" species of wild and domestic birds in Nigeria.

The highly pathogenic avian influenza (H5N1 subtype) recurred in Nigeria after 9 months period of no reported case. A critical look at possible sources of the re-occurrence was desirable. The objective of this study was to determine whether avian influenza viruses were present at reasonably detectable levels (0.5%) in possible "bridge" species of wild and domestic birds. The study was conducted in 8 Nigerian states. A total of 403 birds from 40 species were sampled. Virus isolation was done in embryonated chicken eggs according to standard protocols. The test results were all negative for avian influenza viruses. The overall confidence interval (CI) calculated in R using the exact binomial confidence interval function was 0-0.007406. Tawny Eagle (Aquila rapax) was the lowest sampled 0.3% (1/403) and Red-billed Firefinch (Lagonosticta senegala) the highest 11.7% (47/403). The limitations of the sample size and possibly designing effects on the study, as to make concrete conclusions were acknowledged. Species of wild birds, so identified in the study could be useful in future surveys. Furthermore, multidisciplinary and community oriented approach, blending targeted and passive surveillances was suggested. This approach was envisaged to bring about wider coverage of "bridge" species and clearer insight of their possible roles in avian influenza re-occurrences and spread in Nigeria.

Molecular characterization and epidemiology of the highly pathogenic avian influenza H5N1 in Nigeria.

Avian influenza caused infection and spread throughout Nigeria in 2006. Carcass samples (lung, liver, spleen, heart, trachea and intestine) from the different regions of Nigeria were processed for virus isolation. Infective allantoic fluids were tested for avian influenza viruses (AIV) and Newcastle disease virus using monospecific antisera. Thirty-five isolates were generated and characterized molecularly using the haemagglutinin gene. The molecular analysis indicated that different sublineages of the highly pathogenic avian influenza (HPAI) H5N1 viruses spread throughout Nigeria. We compared the Nigerian isolates with others from Africa and results indicated close similarities between isolates from West Africa and Sudan. Some of the analysed viruses showed genetic drift, and the implications of these for future epidemiology and ecology of avian influenza in Africa require further evaluation. The spread of primary outbreaks was strongly linked to trade (legal and illegal), live bird markets, inappropriate disposal, and poorly implemented control measures. No strong correlation existed between wild birds and HPAI H5N1 in Nigeria.

Serologic and virologic surveillance of avian influenza in Nigeria, 2006-7.

Since January 2006, H5N1 avian influenza has affected Nigeria's poultry population causing enormous loss of resources. The current circulating virus is a potential candidate for pandemic influenza which may severely affect the human and animal population worldwide especially in the resource-poor countries. In this study, we report on our field and laboratory surveillance efforts in Nigeria. A total of 1,821 tissue samples, 8,638 tracheal swabs, 7,976 cloacal swabs and 7,328 avian sera were analysed over a period of two years, with 312 positive results [corrected] We recovered 299 isolates of highly pathogenic avian influenza virus H5N1 mainly from the diagnostic samples of poultry kept in backyard, small scale and free range farms. This finding emphasised the role played by these farming systems in the dissemination of avian influenza in Nigeria and highlights the need for a continued surveillance in humans since human-animal interaction is a key feature in Africa. Furthermore, there is a need for the strengthening of border controls. Since October 2007, there has been no reported and confirmed outbreak of avian influenza in Nigeria.

Control versus no control: options for avian influenza H5N1 in Nigeria.

In January 2006, an outbreak of a highly pathogenic avian influenza (HPAI) was recorded in Nigeria for the first time. This present work describes an estimation of possible costs associated with a vaccination-based control policy added to other measures to restrict HPAI H5N1 virus infections. The evaluations used epidemiological and production data, including budgets necessary for the vaccine acquisition, distribution and administration in arriving at the final costs. Using decision tree and cost benefit analysis the economical benefits for Nigeria and countries with similar veterinary infrastructures, biosecurity and farming systems are calculated. The result indicated that a halting in the continued spread of the virus through effective control measure will be 52 times better than taking no action. This should help policy makers in deciding in favour of vaccination combined with other tools as an effective means of controlling avian influenza H5N1. * Control of HPAI H5N1 will best be understood by policy makers in financial terms. * Effective control through vaccination of poultry is much cheaper and reduces the chances of human zoonoses. * Poultry vaccination combined with other control measures will be the most effective means of control in most developing economies.

Pathogenicity of attenuated peste des petits ruminants virus in sheep and goats.

Progressive loss of virulence for goat kids was noticed when peste des petits ruminants (PPR) virus was passaged in Vero cells. While goats inoculated with the 60th passage suffered from the clinical PPR disease and mortality, goats inoculated with the 80th passage did not show any sign of the disease. If the progressive loss of virulence of the virus with passage continues, it will not be long before a homologous PPR vaccine will be obtained at the National Veterinary Institute, Vom.

Serum-virus simultaneous method of immunisation against peste des petits ruminants.

Peste des petits ruminants (PPR) hyperimmune serum was prepared in cattle and used to protect goats against PPR. The serum was well tolerated by goats and 8 ml of serum was found to protect goats against 4 ml of pooled virulent PPR virus. A durable active immunity was conferred on goats when serum and virulent PPR virus were given simultaneously. Passive immunity lasted for 15 days.