University Diagnostic Lab

Dr. Aamir GhafoorPhD Microbiology (NZ)DirectorUDL, UVAS Lahore

Activit ies 2015 Total Postmortem performed from April 2015-

March 2016= 980 Respiratory Diseases Suspected of H9, IB, ND

= 543 Fowl typhoid suspected = 145 Mycoplasma suspected = 02 Colibacillosis = 700 Coccidiosis =267

2

Culture & Sensitivity

Water Testing- pH range= 7.2-8.5Coliform positive- 40%

3

Bacterial Isolation (%)(Culturing and PCR)

4

Virus Isolation

5

Status of AIV H9 in and around Lahore Results showed that 19% broiler flocks

were affected from Low Pathogenic Avian Influenza Virus (H9) out of the 192 flock

Influenza season in Pakistan starts from December onwards, while, mid-April to mid-August was found to possess highest risk

6

7

Status of NDV in and around Lahore Emerging virulent NDV strains from a new sub-

genotype genotype = VIIi have Panzootic characteristics.

ND outbreaks in three countries Israel, Indonesia & Pakistan are linked by common sub-genotypes.

Ancestral NDV strains from wild birds suggest the existence of unknown reservoirs

On the basis of several full NDV genome analysis of from various regions of the country, data provide the evidence of a novel strain of vNDV (VIIi)

8

9

Sharkpur RoadNovember ,15- UDLND Confirmed Mortality 65%

ArifwalMarch, 16- vet Dx Lab Dr TahirND + AIV H9 Confirmed Mortality 30-55%

KarachiDecember, 16- vet Dx Lab Dr TahirND Confirmed Mortality 55%

Islamabad December, 16- PRI Lab Dr Farhan ND Confirmed Mortality 55%

10

11

12

Lets Think logically! Is the virus changed

Change in cleavage sites Changes in HA gene BUT mostly cross reactive and protective immunity is attained by

regualr vaccination Vaccine and vaccination

Cold chain was a problem in the past but it has been improved Vaccination Practises

Compromised Immune response? Non-uniform growth, fast metabolism, improper ventilation, un-

noticed microenvironment , weather effect

13

HI titers of ND

14

Dry season J.A. NWANTA, S.C. EGEGE, J.K. ALLI-BALOGUN and W.S EZEMA (2008). Evaluation of prevalence and

seasonality of Newcastle disease in chicken in Kaduna, Nigeria. World's Poultry Science Journal, 64, pp 416-423. doi:10.1017/S0043933908000147.

U. Musa, P.A. Abdu, I.I. Dafwang, J.U. Umoh, L. Sa`idu, U.M. Mera and J.A. Edache, 2009. Seroprevalence, Seasonal Occurrence and Clinical Manifestation of Newcastle Disease in Rural Household Chickens in Plateau State, Nigeria. International Journal of Poultry Science, 8: 200-204.

15

• Pakistan (n= 16); poultry swabs samples from 16 outbreaks in different regions of the country; Oct., 2011 - March 2012

• Indonesia (n =14); diagnostic swab samples from live bird markets or poultry handling facilities.

• Israel (n = 33); diagnostic cloacal & tracheal swabs from poultry & pet birds [Kimron Veterinary Institute]

• NDV strains (n= 3) cockatoo/Indonesia/87-36724-524/1988; lory/Indonesia/88-08989-523/1988 & parrot/Indonesia/C300 (19625)-520/1976; repository of USDA/SEPRL (ancestral)

• ICPI (~14) & MDT (~32): most velogens, some mesogens. “RRQKRF”

Two ClassesClass II: contains 18 genotypesGenotype V, VI, VII, XIII has sub-genotypes Genotype VII: divided into 8 sub-genotypes

• Pigeons and Cormorants: well-recognized reservoir of genotype V & VI

• Reservoir for other ND strains (VII, XII, XIII, XVIII) is unknown. • Failure to control ND with vaccination: where the virus is

endemic

Status of FAdV in and around Lahore Phylogenetic analysis of 10 recently isolated fowl

adenovirus (FAdV) isolates associated with Inclusion Body Hepatitis (IBH) in Punjab during 2014-2015.

Two different types of serotype, FAdV-4 and FAdV-11 were identified.

Seven Pakistani Newly isolates FAdV strains (SFR1D-SFR7D) were closely related & clustered into serotype 11 in species D.

Three FAdV isolates (AW1-AW3) were clustered into serotype 4 in species C.

18

UDL Use of molecular biology techniques

Endpoint PCR Real-Time PCR Sequencing

In addition to positive and negative controls, use of internal controls

Latest and improved methodologies of microbial isolations

Improved Quality Control Our integration with on-going research projects

19

Acknowledgments ND and FAdV data

USDA /CRDF Newcastle Disease NDV 31063 project

PARB funded project “Diagnostic Survey for Newcastle disease (ND) in Poultry” by

Dr. Shafqat Fatima Rehmani QOL UVAS AIV H9 data

Avian Influenza project (Dr Tahir Yaqoob) Department of Microbiology UVAS

20

Thanks

21