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Annual Technical Report
Fiscal year 2071/072 (2014/015)
Government of Nepal
Ministry of Livestock Development
Department of Livestock Services
Directorate of Animal Health
Central Veterinary Laboratory
Tripureshwor, Kathmandu, Nepal Phone: +977-1-4212143, 4261938 Fax: +977-1-4261867 Email: cvl@cvl.gov.np Website: www.cvl.gov.np CONT Foreword
I am very much pleased to present this annual technical report of our laboratory for the fiscal year 2071/072 (2014/015). This issue includes activities and progress report of Central Veterinary Laboratory (CVL), National Avian Disease Investigation Laboratory (NADIL) and five Regional Veterinary Laboratories (RVLs). Detection of disease is the foundation for disease control, prevention and eradication. Early and accurate diagnosis of diseases can only be assured in fully equipped laboratories that have range of standardized diagnostic reagents with trained human resource. Nepal being the member of World Trade Organization (WTO) has to implement SPS measures on scientific merit. Laboratory based diagnosis is of paramount importance for implementing the SPS measures in the territory. Therefore, the role of national and regional veterinary laboratories including basic laboratories situated in the districts is very important in disease diagnosis of major livestock species in the country. Still there is a scope to uplift the standards and quality assurance of the laboratory diagnosis conducted by existing facilities. With the establishment of molecular diagnostic techniques at the CVL, routine molecular diagnosis of Avian Influenza, ND, and IBD is performed. Similarly, the ELISA, IFAT, HA/HI, AGPT, virus isolation and characterization and other routine diagnostic tests are used for the diagnosis of various livestock and poultry diseases. We are putting our efforts to upgrade CVL, RVLs and NADIL to provide reliable and prompt diagnostic services all over the country. We already have good co-ordination between regional labs and basic labs and the sample flow in the national reference laboratories is routinely being done. I would like to extend my sincere thanks to Food and Agriculture Organization (FAO) for their support in diagnostic reagents. Likewise, technical support of Australian Animal Health Laboratory (AAHAL) and Veterinary Laboratory Agency (VLA), WEY bridge, U.K. for molecular characterization of different viruses is highly acknowledged. Sincere efforts of all RVLs and NADIL family for providing information required for this publication is really appreciable. Finally, I express my personal appreciation and sincere thanks to all the staffs of CVL who worked hard to shape the annual technical report in this form. I would be grateful if you provide any suggestions for the improvement of the future issue of this publication.
Dr. Samjhana Kafle (Pandey)

Chief Veterinary Officer
Central Veterinary Laboratory
Tripureshwor, Kathmandu, Nepal
CONTENTS
S.No Title
A Central Veterinary Laboratory 3 Organization Structure 4 Laboratory Services B National Avian Disease Investigation Laboratory, Chitwan 3 Organization Structure 4 Laboratory Services C Regional Veterinary Laboratory, Biratnagar 3 Organization Structure 4 Laboratory Services 5 Epidemiological parameter of Eastern Development Region of Nepal D Regional Veterinary Laboratory, Janakpur 3 Organization Structure 4 Laboratory Services 5 Sample sent to CVL for further Investigation in F/Y 2071/072 E Regional Veterinary Laboratory, Pokhara 3 Laboratory Services F Regional Veterinary Laboratory, Surkhet 3 Organization Structure 4 Laboratory Services 5 Epidemic Investigation G Regional Veterinary Laboratory, Dhangadhi 3 Organization Structure 4 Laboratory Services 5 Investigation programs A. CENTRAL VETERINARY LABORATORY KATHMANDU

1. Introduction:

Central Veterinary Laboratory (CVL) focuses program with the objective of securing
healthy national flocks of animals and birds throughout the nation by mitigating the
occurrence of diseases of livestock and poultry. CVL also works on epidemic
investigation as well as surveillance and investigation on various diseases in its approved
annual program. The direct benefit of the performance of various laboratories has been
experienced in the field of veterinary medical care based on valid laboratory test results.
To achieve these multidimensional activities, CVL works with a series of laboratory test
procedures through its various laboratory sections; Pathology, Parasitology,
Microbiology, Serology, Laboratory management and teaching lab and Molecular
Biology with a considerable progress in the later. At present the molecular based
diagnosis of avian influenza is in routine use.
CVL has standard Operating Procedures, test protocols and quality guideline manual.
CVL is gradually practicing the biosafety/ biosecurity measures so that good laboratory
practice is followed in our all the diagnostic laboratories. CVL has already been adopting
test verification system through international reference laboratories which will help in the
accreditation of CVL for international certification as well.
To provide diagnostic facilities throughout the country, CVL works through its five
Regional Veterinary Laboratories (RVLs) located one in each of the development regions
of the nation; eastern (Biratnagar), central (Janakpur), western (Pokhara), mid-western
(Surkhet) and far-western (Dhangadhi) as well as through National Avian Disease
Investigation Laboratory (NADIL) located in Chitwan. To provide the diagnostic
services smoothly throughout the nation, 15 basic laboratories have been established in
15 District Livestock Service Offices (DLSOs) namely, Illam, Jhapa, Saptari, Sarlahi,
Rautahat, Parsa, Makawanpur, Kabhrepalanchowk, Chitwan, Rupandehi, Dang, Banke,
Jumla, Dadeldhura and Kanchanpur, and 60 primary laboratories one in each DLSOs of
other districts. The basic laboratories are capable to perform parasitological
examinations, microbial culture and antibiotic sensitivity test. Specimens that cannot be
processed in the aforementioned laboratories due to insufficient facilities and expertise
are referred to the CVL. In this way, CVL works as a reference veterinary laboratory in
Nepal.

2. Objectives:

The role of veterinary laboratory system has become dynamic in the advent of food safety issues, economic liberalization and trade globalization. Nepal joined as a WTO member in 2004. Therefore, Nepal follows the guidelines provided by Office International des Epizootics (OIE) for the provision of Sanitary and Phytosanitary (SPS) agreement under WTO that seeks scientific procedures and evidences in the course of disease diagnosis as well as production chain. The roles of veterinary diagnostic laboratories are now therefore expanded and challenging in the new context. Moreover, CVL works with the following objectives in the country.  Provide laboratory diagnostic services in the country in the area of animal health and veterinary public health.  Acts as a national veterinary reference laboratory.  Conduct epidemiological disease investigation & laboratory diagnosis of livestock and poultry disease.  Support the national disease control and surveillance programs.  Acquire, adopt, upgrade and disseminate different laboratory diagnostic test methodologies for Livestock and poultry diseases.  Assist Animal Health Directorate (AHD) in the animal health policy development and formulation of animal disease control and eradication programs.  Extend & disseminate information concerning livestock and poultry disease controls.  Collaborate with international reference laboratories & institutions on veterinary laboratory diagnosis.  Assist in implementation of national epidemic control strategies.  Conduct laboratory diagnosis techniques training for the veterinarians and veterinary paraprofessionals.  Strengthen and coordinate regional and district level veterinary laboratories.  To achieve the aforementioned objectives, there are a series of approved annual activities carried out by different laboratory sections of the CVL, five RVLs and NADIL.


3. Organization Structure:
Table: List of Staffs working at CVL (At the end of F/Y 2071/072)
Position
Dr. Samjhana Kumari Kafle Chief Veterinary Officer Dr. Pragya Koirala Senior Veterinary Officer Dr. Rajesh Yadav Senior Veterinary Officer Veterinary Officer Mrs. Gauri Devi Thapa Veterinary Officer Mr. Purna Bahadur Budha Veterinary Officer Mr. Bal Bahadur Kunwar Veterinary Officer Mr. Prakash Devkota Veterinary Officer Dr. Manju Maharjan Veterinary Officer Dr. Chanda Shrestha Veterinary Officer Mr Bhakta Bahadur Animal Health Technician Mr. Shyam Sundar Yadav Animal Health Technician Animal Health Technician Mr. Tul Bahadur Rai Animal Health Technician Mr. Mithilesh Kumar Karna Animal Health Technician Assistant Animal Health Mr. Purna Maharjan Assistant Animal Health Mr. Bhimsen Adhikari Assistant Animal Health Mr. Bal Kumar Rai Senior clerk (Typist) Mr. Buddhi Bahadur Lama Ms. Prem Kumari Sherpa Clerk (Kharidar) Mr. Machakaji Maharjan Mr. Dipesh Rana Magar Mr. Rajan Adhikari Mrs. Bheema Acharya Mr. Chandra Bahadur Rana Office Helper (Contract) Table : Annual program & progress report of CVL in the Fiscal Year 2071/072
Activities
Progress
Fecal examination EPG counts of parasites Skin scrapping examination Pathology
Clinical hematological examination Bio-Chemical examination Urine examination Post-Mortem Examination Hitopathological examination Isolation and Identification of Bacteria Diagnosis of Mycoplasma in goats Sample collection, Isolation and Identification of Fungus Virology
Sample collection of virological Investigation of PPR outbreak PPR Diagnosis by ELISA and Pen site Serology
PPR Sero-monitoring for National PPR control program Poultry sample collection and examination for salmonella and Molecular Diagnosis
Molecular Diagnostic examination for Gene sequencer maintenance Dispatch of sample to international reference laboratories Disease surveillance and investigation
Emergency disease investigation team (EDIT) deploying Sample collection and examination of PRRS disease in pig Activities
Progress
Investigation of livestock and poultry Zoonotic disease Investigation
Sample collection and examination for Rabies Sample collection and examination for Food security
Clinical mastitis Investigation in Dairy Staff Development
Training on Laboratory Technology (30 Days) for mid Level Technicians Purchase of technical issues and journals Interaction programs
Participation on Regional laboratory workshops Laboratory technology and program planning workshop Participation in Regional review workshops in Regional Directorates Laboratory Monitoring
Monitoring and evaluation of Regional and basic laboratories Publication of Laboratory Technical Laboratory Management
Health examination of staffs Teaching lab management Management of Serum Bank Lab animal management 4. Laboratory Services:
4.1 Parasitology section:
The parasitology section is involved in routine examination of different types of internal and external Parasites of animals and birds, besides of this, the parasitology section is also involved in the investigations of adverse effect on domesticated animals and poultry health and production. For the investigation and diagnosis of parasites, fecal samples, blood samples and skin scrapings samples are collected and examined by adopting standard veterinary laboratory protocols. As a routine examination or diagnosis of parasites, direct smear method, Sedimentation method and the floatation methods are commonly used as per the guidelines for detection of parasites. These methods are adopted for both of the quantitative and qualitative
detection of parasites. The quantitative method is performed by Mc Master's slide for the
detection of numbers of eggs, of the gastrointestinal parasites in a gram of faces.
The main objectives of Parasitological section is to, provide day to day diagnostic service
to the farmers who come to this Laboratory with samples for parasitic diagnosis.
Likewise, investigations in different parts of country are also conducted by CVL as a
regular program.

Parasites identified during the fiscal year 2071/72
In the fiscal year 2071/72, a total of 837 fecal samples of cattle, buffaloes, sheep, goats,
dog, poultry and other species of animals were examined in this section. Among them
100 samples were collected by CVL staff for performing the EPG count as per the CVL's
regular programmer. Out of 837 fecal samples, 331 samples were found to be positive for
different species of internal parasites. Moreover, 63 samples of skin scraping were also
collected from Dogs for the detection of Mites. The results of examinations are in the
tables below as a whole.
Table: Different types of parasites detected in the fecal samples of livestock and
poultry species.
Name of parasites
Total samples collected
Total positive samples
Paramphitomum sp. Table: Month wise distribution of parasites examined in fecal samples.
Number of different Species of Parasites and their percentage
Monthsz Sample
Trichuris Ascaris Capilllaria Coccidia 4 (6.5%) 7 (11.4%) 15 (18.7%) 3 (3.7%) 6 (11.1%) 2 (3.7%) 22 (34.3%) 1 (1.5%) 72 (38%) 15 (7.9%) 8 (4.2%) 13 (6.8%) 3 (1.6%) 11 (21.1%) 1 (1.9%) 2 (3.8%) 13 (24%) 3 (5.5%) 8 (14.8%) 2 (3.7%) 1 (1.8%) 168 (20%) 127 (15.1%) 216 (25.8%) 27 (3.2%) 16 (1.9%) 11 x1.3%) 11 (1.3%) 14 (1.6%) 4.2 Pathology section:
Post-mortem examination, hematology, histopathology & clinical biochemistry are major
areas under the pathology section in CVL. Mostly the section receives specimens from all
over the country either directly or through the respective RVLs and NADIL. Besides this,
the District Livestock Service offices (DLSOs), veterinary practitioners and livestock and
poultry farms as well as farmers deliver specimens for the purpose of disease diagnosis.
Necropsy examination plays very important role in the diagnosis of diseases in different
species of animals. Necropsy examination is the first step of disease diagnosis in case of
dead animals. The history, clinical findings, epidemiological surveillance information is
also helpful to necropsy examination for diagnosis of diseases.

4.2.1 Post-mortem examination:

During the fiscal year 2071/072 all together 3840 animals/poultry carcass were brought
for post-mortem examination at CVL. The detail month-wise and species-wise
distribution is shown in the tables below.
Table: Month-wise distribution of postmortem examination and pathological
conditions diagnosed for species other than Avian at CVL.
Sum Total
The table above depicts that very few carcasses of species other than avian were registered at CVL for Postmortem examination. Table: Month-wise distribution of postmortem examination and pathological
conditions diagnosed for Avian species at CVL.
Peritonitis/Salpingitis Encephalomyelitis Sum Total
The table above clearly shows that maximum numbers of avian carcasses were examined at CVL in the month of Bhadra followed by Shrawan and Chaitra. Most frequently encountered pathological conditions in avian species were Colibacillosis, cCRD, Infectious Bursal Disease and Mycotoxicosis. 4.2.2 Heamatology and Biochemistry:
A total of 633 blood samples were tested in FY 2071/072 for blood analysis. Out of 512
cattle blood samples examined 42 were positive for Babesia spp., 16 for Anaplasma spp.
Likewise, out of 23 buffalo blood samples, 4 were positive for Babesia spp and 2 for
Anaplasma spp. Similarly, out of 66 Dog blood samples, 6 were positive for Babesia spp.
and 1 for Tripanosome spp. Furthermore, 8 Equine blood samples were examined only
one sample was positive for Trypanosoma spp.
Table: Details of blood samples tested.
Blood protozoa positive
Buffaloes
4 Babesia, 2 Anaplasma 1 Tripanosoma sps Goat/Sheep
Table: Urine and blood samples tested for biochemical parameters.
Type of Sample tested
Buffaloes
Phosphorous
Magnesium
4.2.3 Histopathological examination:
Post-mortem examination provides a tentative diagnosis of a disease and the
histopathology technique always lays a presumptive diagnosis. In this way, it is essential
to circulate necropsy and sometimes biopsy samples through various laboratory
disciplines. Histopathology section processes the samples and provides the result within
7-10 days period by wax techniques method.
Histopathology unit received a total of 62 samples from different animal species during
the fiscal year 2071/072. Out of 62 samples examined, 13 (20.96%) samples were found
abnormal and 49 (79.04%) samples were found not abnormal detected (NAD). Those 13
avian samples found positive for various histopathological conditions were Infectious
Bursal Disease (5), Coccidiosis (2), Nephritis (2), Pneumonia (2) and Hepatitis (2) were
the major indicative disease conditions.
4.3 Microbiology section:
4.3.1 Bacteriology and mycology unit:
The bacteriology unit receives samples from various sources such as farmers, Central
Veterinary Hospital, referral samples from private clinics, Regional Veterinary
Laboratories and directly from the field as well DLSO's. Various samples like milk,
urine, tissues, water, nasal swabs, ear swabs and skin scrapping were received in this unit in fiscal year 2071/72. Different bacterial species were isolated from specimens as given in the following table. Table: Bacterial species isolated in the laboratory.
Number of
Bacterial species
isolated
Escherichia coli Staphylococcus spp. Streptococcus spp. Micrococcus spp. Haemophillus spp. Enterobacter spp. Pasteurella multocida Likewise, a total of 49 fungal samples were received in this unit for mycological culture and among these 11 samples were found positive for fungal growth. Fungus isolate from pathological lesion of PM sample and poultry feed were penicillum spp. Table: Fungal species isolated in the laboratory.
Fungal species
Number of isolated
Moreover, a total of 352 bovine samples and 15 caprine sample were received in this unit for CMT and bacteriological culture and among these 7 samples of bovine samples and 3 samples of caprine samples were found negative for CMT test and no growth on bacteriological culture. Table: Bacterial species isolated in the milk samples of CMT positive cases.
Bacterial species
Number of isolated
Escherichia coli Staphylococcus spp. Streptococcus spp. Micrococcus spp. any of the bacterial isolates from the general samples received were subjected to antibiotic sensitivity test. Among the antibiotic Gentamicin, Tetracycline, Ciprofloxacin and Azithromycin were sensitive for E. Coli, Staphylococcus, Salmonella, Streptococcus, Bacillus, Shigella species. Table: Antibiotic sensitivity test result in general samples.
Bacterial
Antibiotic sensitivity percentage
Note:
G-Gentamycin, T-Tetracycline, Am-Amoxycillin, A-Ampicillin, Azm-Azithromycin,
N-Neomycin, Ex-Enrofloxacillin, Ak-Amikacin, Le-Levofloxacin, Cf-cephalexin,
Nf-Norfloxacin, erythro-Erythromycin, Cl-chloramphenicol, Do-doxycycline, B-bacitracin
Furthermore, bacterial isolates from the milk sample received from mastitis cases were subjected
to antibiotic sensitivity test. Among the antibiotic Gentamicin, Tetracycline, Ciprofloxacin,
Azithromycin were sensitive for the treatment of cattle in case of mastitis disease for the given
organism.
Table: Antibiotic sensitivity test result in mastitis positive cases.
Bacterial
Antibiotic sensitivity percentage
N. species
Escherichia coli Note:
G-Gentamycin, T-Tetracycline, Cip-Ciprofloxacin, Am-Amoxycillin, A-Ampicillin, Azm-
Azithromycin, N-Neomycin, Ex-Enrofloxacillin, Of-Orfloxacin, C-colistin,
Ak-Amikacin, Le-Levofloxacin, Cf-cephalexin, Nf-Norfloxacin, erythro-Erythromycin, Cl-
chloramphenicol

4.3.2 Virology unit:
Virology unit is responsible for the diagnosis and monitoring of viral diseases . Most of
the samples are submitted to this unit by the post mortem unit of CVL, Regional
Veterinary Laboratories, National Avian Laboratory, Central Veterinary Hospital,
District Livestock Service Offices, quarantine check posts, private clinicians, farmers and
staffs of CVL during disease outbreak investigations as well as for routine diagnosis. The
unit has a capacity of Competitive Enzyme Linked Immunosorbent Assey (ELISA),
Flourescent Antibody Test, Virus isolation and identification, Plate agglutination test and
rapid test. Rapid test is used for the initial diagnosis of New Castle Disease, Infectious
Bursal Disease, Avian Influenza and Rabies. For the further confirmative diagnosis of
Avian Influenza the samples are sent to the molecular section for Polymerase Chain
Reaction (PCR). Likewise for the diagnosis of rabies, rapid test kit method is followed by
Negri body test, FAT and mouse inoculation test. In case of New Castle disease HA/HI is done. Progress report of virological investigation of various diseases in animals and birds during 071/72 is as follows: Table: Month-wise distribution of Rabies.
Canine Feline
Rabbit Mice
Positive Negative
Of the total samples received for the diagnosis of rabies 49.09% of the samples were found to be positive. The samples were initially tested with rapid antigen test kit method which was followed by Negri body test, FAT and finally mouse inoculation test. Table: Month-wise distribution of diagnosis of poultry disease by rapid test kit.
New Castle disease Infectious Bursal Disease In the fiscal year 2071/72 a total of 118 tracheal samples were received for the diagnosis of New Castle disease which were initially tested with the rapid antigen detection test kit followed by HA/HI test. Of 118 samples 15.25% were found to be positive. From the post mortem unit 170 samples were submitted for Avian Influenza diagnosis of which 14.7% were found to be positive on rapid test. Also, a total sample of 172 tracheal, cloacal swab and brain samples were received from NAL, RVLs and DLSOs. The positive samples along with 10% of the negative samples were sent to molecular unit for the identification of subtypes H9 and H5. Similarly, for Infectious Bursal disease only
rapid test kit method is implied. Of the total samples 64.07% were found to be positive.
Likewise, a total of 110 nasal and ocular samples were received for PPR antigen test.
Only 28.18% were found to be positive. The samples were initially diagnosed through
penside test followed by C-ELISA.

41

4.4 Serology section:
Serology section of CVL performs different serological tests for the diagnosis,
monitoring and surveillance of animal diseases mainly associated with viral and bacterial
infection. Most of the samples are submitted to this unit by Regional Veterinary
Laboratories, District Livestock Service Offices, Quarantine Check-posts, private
practitioner, farmers and staff of CVL during disease outbreak investigations well as
routine diagnosis. This section possess capacity and facility of Competitive Enzyme
Linked Immunosorbent Assay (ELISA), IgM capture ELISA, Indirect ELISA, Tube
agglutination Test, Agar-Gel Immuno-Diffusion(AGID) test, Plate agglutination test, A
solid phase immune assay (Immuno- comb) and rapid tests. Progress report of
Serological investigation of various diseases in animals and birds during 2071/072 is as
follows
Table : Peste-des Petitis Ruminant (PPR), C-ELISA Antibody Test Results
(Outbreak Samples)
Test Results
Percentage
Districts
No. of sample
Positive (%)
Positive
Negative
A total of 269 serum samples of goats from outbreaks areas were tested, 59.11 % were positive for PPR antibody. The result shows that the PPR antibody positive percentage was found 100% Dadeldhura, Makwanpur and Syanja districts. Table : Peste-des Petitis Ruminant (PPR) Seromonitoring C-ELISA
Antibody Test Results
Test Results
Percentage
Districts
No. of sample
Positive (%)
Positive
Negative
A total 3939 serum samples of vaccinated goats were tested. Out of those samples 57.20% were found positive for PPR antibody. The result shows that the PPR antibody positive percentage was found highest in Terhathum (91.09%) followed by Dolkha and Sindhupalchowk districts.The antibody positive percentage found in the serum samples of Doti, Taplejung, Dadeldhura, Solukhumbu, Ilam, Siraha, Okhaldhunga, Udaypur and Sankhuwasabha were not satisfctory . The low antibody positive percentage might be due to either sampling error or cold chain problem or the samples were collected earlier. Table: Antibody Test Result of Brucellosis Antibody by ELISA
Test Results
Districts
No. of sample
Positive
Negative
A total of 644 serum samples from cattle (608) and buffalo (36) were tested for Brucellosis antibody by ELISA method and none of samples were found positive. Likewise 105 serum samples from different species was tested by plate agglutination test with negative results. Table: Antibody Test Results of Brucellosis PAT
Brucella abortus (PAT)
Districts
Toatal sample
Positive
Negative
Table: Antibody Test Results of Salmonella pullorum and Mycoplasma gallisepticum
in poultry serum by using Plate Agglutination Test (PAT)
Salmonella pullorum (PAT)
Mycoplasma gallisepticum (PAT)
Districts
Total Tested
Total Tested
Positive Negative
Positive Negative
A total 201 number of poultry serum samples were tested for Salmonella pullorum antibody by PAT method and 37 were found positive. Similarly, 201 number of poultry serum samples were tested for Mycoplasma gallisepticum antibody by PAT method and 19 were found positive. Table : Antibody Test Results of Porcine Reproductive and Respiratory Syndrome
Virus (PRRS) in swine serum by using ELISA
Test Results
Districts
No. of sample
Positive
Negative
A total of 411 serum samples of swine were tested for Porcine Reproductive and Respiratory Syndrome (PRRS) Virus Antibody by ELISA method and 28 samples were found positive. Table: Avian Encephalomyelitis Virus Antigen test result by using ELISA
Test Results
Districts
No. of sample
Positive
Negative
Kathmandu Valley A total 91 serum samples of poultry were tested for Avian Encephalomyelitis Virus Antibody by ELISA method. The samples were received from Kathmandu Valley. Out of those samples 81 samples were found positive. Table: Avian Influenza Flu A Virus Antibody Test Results in poultry serum by
No. of sample
Test Results
Districts
Positive
Negative
A total 2535 serum sample of poultry (duck/chicken) was tested for Avian Influenza Flu A Virus Antibody by ELISA method. The samples were received from high risk districts like Morang, Dhadhing, Saptari, Jhapa, Chitwan districts etc. Out of those samples 108 samples were found positive with a overall positivity of 4.24%. 4.5 Molecular Biology section:
Molecular tools are increasingly important in modern animal disease research. In view of
this Central Veterinary Laboratory (CVL) has established molecular biology unit in 2003.
This laboratory has started to diagnose disease especially avian influenza by using RT-
PCR technique. This technique is also useful for the epidemiological study of diseases
and understanding of the genetic relationships between and within the species of
infection organisms.
Table: Test results of Molecular diagnosis of avian influenza virus from suspected
samples by PCR Method
Total No. of
Result (Positive for
District
Subtype H9)
In the Fiscal year 2071/72, a total of 97 tracheal swab samples different types of avian species were tested for Avian Influenza (Mgene and other Subtypes) by molecular diagnosis method (PCR method). Out of those samples 54 samples were found positive for avian influenza subtype H9. During this period, samples were received from 18 districts like Bhaktapur, Lalitpur, Kathmandu, Dhadhing, Kaski, Makwanpur, Mgyadi, Kavre, Sunsari etc. districts. The highest numbers of samples were received from Kathmandu. Like- wise, a total of 91 pooled samples (tracheal swab and environment samples) collected from 9 districts were analyzed by using Real Time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCR) technique for Matrix (M) Gene. Only 5 samples were found positive for M gene and subtype H9. Those samples were collected from different location of poultry slaughter place or live bird market (LMB). Table: Laboratory test results of Samples from surveillance sites- Live Bird Market
(LBMs) and wild bird interface
Total Sample Tested
Surveillance Districts
(Pooled TS/ES)
B. NATIONAL AVIAN LABORATORY, BHARATPUR, CHITWAN

1. Introduction:
National Avian Disease Investigation Laboratory (NADIL) was established in 2004
under the Department of Livestock Service. This is the first commodity (Poultry) specific
lab in country's Livestock Department. It works with direct supervision of Central
Veterinary Laboratory, Tripureshwor. The Laboratory was proposed during ninth five
year plan to address problem related to privately evolving poultry industries in the nation.
So, that it could diagnose earlier problems regarding poultry and protect the farmer from
the losses. Therefore its major objective is to provide essential laboratory diagnostic
services for the avian diseases so it can facilitate the fast growing commercial poultry
industry in the country. More than 60 percent of the total population and the activities
concerned with poultry are concentrated in Chitwan. Poultry sector has about 4 percent
contribution in National economy through poultry and its product. Currently there are 62
hatcheries' and 48 feed industries supporting commercial broilers and layer farming
business in Chitwan and major cities of the country. This laboratory provides diagnostic
service through several units, such as post mortem, Sample collection, histopathology,
parasitology, bacteriology, virology, serology and molecular biology unit.
2. Objectives:
 To diagnose the major poultry diseases by adopting standard laboratory  Isolation, characterization and preservation of different bacteria, virus, fungi, and protozoa causing disease in poultry.  To develop the laboratory as a centre for the avian disease diagnosis.  To assist in national poultry disease surveillance and control programs.  To assess the immunological status of major viral diseases in commercial flock of Nepal through test like ELISA and HA/HI.  Conduct HPAI laboratory diagnosis using real time PCR technique. Organization Structure:
Table: Man power Situation during F/Y 2071/072
Approved
Type of the post
Fulfilled
Technical (Officer)
Senior Veterinary Veterinary Officer Technical (Non-Officer)
Total Technical Staffs
Approved
Type of the post
Fulfilled
Non-Technical (Non-Officers)
Total Administrative staffs
Grand Total
Table: Annual program and summary of progress of fiscal year 2071/072
Programmes and Activities
Progress
Laboratory Services
Parasites Identification Postmortem Examination Histopathological Identification Microbiological Tests
Bacteria Culture & Antibiotic Sensitivity Test Bacteria Stain Preservation Fungal Culture & Identification Virological Test
Virus Isolation & Identification Rapid Test(ND,IBD,AI) Serological Test
PAT For Mycoplasma PAT For Salmonella Molecular Diagnosis
Sample Collection , Examin & Disease Investigation &
Migratory birds (Surveillance, Poultry Disease Diagnosis By Immunocomb ELISA(ND,IBD, Bacterial AST Searing & Epidemiological Reporting
Monthly Epidemic Reporting Monitoring & Supervision
District Lab. Monitoring Training & Workshops
Programmes and Activities
Progress
Participation in Central & Other Workshops In Poultry Entrepreneurships Assistant Level (3 days Sample Publication
Preparation Annual Technical Lab Animal Management
Sheep management for lab 4. Laboratory Services:
Different activities of laboratory were performed by various units. The details of the unit
wise technical progress are presented separately under various headings as below.
4.1 Parasitology unit:
During the F/Y 2071/72 among various fecal samples examined a total of 399 samples
were found positive. Among them 80 percent sample were positive for Coccidia followed
by Ascaria (17%) and intestinal Tapeworms (3%).
Identified Internal parasite

4.2 Bacteriology Unit:
Suspected samples, during post mortem examination were collected and brought to
bacteriology unit for bacterial culture. Total 237 samples were collected and processed,
out of which 122 samples were showed bacterial growth. Those grown micro-organisms
were identified on the basis of their colony structure and growing character in culture
media. Among them 51 percent of the bacterial isolates were identified as E. Coli
followed by Samonella (37 %), Pasteurella sp. (8%) and Pseudomonas sp. (4%).
Table: Month-wise distribution of bacterial isolates at NAL during 2071/072
No of Samples
Salmonella
Pasteurella
Pseudomonas sp.
Mangshir
Sum Total
Bacteria Isolated at NAL

4.3 Post mortem Unit:
Clinical and post mortem examination of sick and dead birds is the major worked of this
unit. Since the establishment of this laboratory, poultry cases are registered at an
increasing rate, which might be to its diagnostic capacity. The suspected samples of
tracheal and cloacal swabs were taken and tested for Avian Influenza, Ranikhet and
Infectious Bursal Diseases by using rapid test kits. Blood sample were taken and sent to
serology unit for further investigation. Diagnosis was also made by observing
pathognomic and allied lesion during the postmortem examination.
Total 41 type's of pathological conditions were diagnosed in this fiscal year. Among them Fowl cholera dominates of all the pathological conditions followed by CCRD, Salmonellosis, Collibacillosis, Mycotoxicosis etc. Altogether 16 major pathological conditions were diagnosed in the laboratory including Ascites, LPAI, Omphalitis, Coccidiosis, LS and worms. Trend of total number of PM examination over last seven years
Fiscal Year
Up to 16 Major Poultry Disease in NAL by PM Diagnosis Fow l cholera+ Mycotoxine Figure: Districts wise enrollments of bird for Post-mortem
examination

Present Service Coverage of NAL
Total 9195 cases during this year were enrolled from different 18 districts. The samples were from eastern Mahottari to Kailali in the western part of Nepal. The maximum number of were from Chitwan (70%) followed by Nawalparasi (15%), Makawanpur (7%), Tanahun (2%), Gorkha (2%), Dhading (2%) and rest of the districts contributed less than one percent. Likewise, highest number of birds were enrolled from Commercial Broilers (74%) followed by Commercial Layers (22%), Parent Stock (4%) , Giriraja, Backyard Poultry, Ducks and Crows. Status of birds registered in NAL for 1 Commercial Broilers 2 Commercial Layers 3 Broiler parents 4 Backyard chicken 6 Backyard Ducks 4.4 Virology/Serology Unit:
Various tests were performed at Virology/Serology unit during fiscal year 2071/2072.
Out of 315 samples tested for Samnonella sp. with PAT, 93 samples were positive.
Similarly PAT test was also performed for Mycoplasma infection in 218 samples with 55
samples recorded as positive. Avian Influenza flue A was tested by rapid test kit in 634
samples resulting 98 were positive. Likewise, ND (190 samples) and IBD (318 samples)
were tested by rapid test with 44 and 186 positive for ND and IBD respectively.
Moreover, Immunocomb test was performed for ND, IBD and IB. Among them 76 were
found positive for ND, 84 positive for IBD and 28 were IB positive from each 110
samples. Furthermore, a total 260 ELISA test were done for type A avian influenza with
98 samples positive.
Table: Various screening tests employed for diagnosis of major poultry diseases.
Name of the diagnostic
No. of samples
No. of positive
Plate Agglutination Test Plate Agglutination Test Rapid Test (AIV) Rapid Test (IBD) Immunocomb (IBD) ELISA ( Avian Influenza Sum Total

4.5 Molecular biology Unit:
Altogether 121 samples were tested for confirmation of Avian Influenza type A virus at
molecular biology unit of the laboratory. The molecular technique employed in the
laboratory was reverse transcriptase polymerase chain reaction (RT-PCR). Among all the
121 samples tested for the confirmation on 70 samples were found positive for Avian
Influenza type A Virus.
4.6 Histopathology unit:
Histological sections of different samples from post mortem unit were made to
investigate the poultry diseases. Total 281 slides were made and 93 slides were
interpreted with various pathological conditions. Among them major slides were
diagnosed as Coccidiosis followed by Marek's Disease (11 slides), Mycotoxicosis,
Hepatitis, Pneumonia, Enteritis and IBD from detailed histopathological study by
experts.
C. REGIONAL VETERINARY LABORATORY, BIRATNAGAR
(EASTERN REGION)
1. Introduction:
Regional Veterinary Laboratory (RVL) has been situated in sub-metropolitan city,
Biratnagar-17, of eastern Nepal and was established in the fiscal year 1988/1989 AD. But
until1990/1991, the laboratory was not functional and could not perform its activities as
per objectives due to lack of manpower, necessary equipments and frequent changes in
organizational structure. From fiscal year 1991/19992, the RVL has its separate identity.
There was provision of manpower and other logistics. The program was launched as per
objectives.
The working area of this RVL is all districts of Eastern Development Region (EDR). In
this eastern region, there are three zones (Mechi, Koshi and Sagarmatha) and 16 districts.
Geographically, the region is divided into three eco-zones (high hills, mid hills and terai).

High hills:
This eco- zones lies in the northern part of the region covering Taplejung district of
Mechi zone, Sankhuwasabha district of Koshi zone and Solukhumbu district of
Sagarmatha zone. Livestock rearing is the main occupation of the farmers in this region.
Yak/Nak, chauri, sheep and goat are being reared in this region.

Mid-hills:
This region falls between high hills on its north and tarai at the south. Panchthar, Illam,
Dhankuta, Terahthum, Bhojpur, Okhaladhunga, Khotang, and Udaypur districts are under
this eco-zone. Farmers follow mixed farming system and agro-based livestock industries
are their main occupation. Cattle, buffalo, swine, goat are being reared in this region.
Poultry and rabbit farming are also popular among the farmers

Terai:
Jhapa, Morang, Sunsari, Saptari and Siraha districts of Nepal are under terai eco-zone.
Though traditional system of livestock rearing is followed in this region, in recent years,
poultry, dairy industries and piggery are being commercialized especially in east-west
highway corridor.
To provide proper laboratory diagnosis and improve in the quality of veterinary services,
the government has established five regional laboratories, one in each development
region of the country. Primary laboratories in 55 and basic laboratories in 15 districts
have been established to improve and upgrade existing disease diagnosis system.
2. Objectives:
 To provide prompt and efficient disease diagnostic services to the farmers of  To investigate and diagnose the epidemics in the region.  To assist and support DLSOs in disease diagnosis and epidemic control.  To supervise and assist in diagnostic services to basic and primary laboratories situated in DLSOs of the region.  To collect, analyze and predict the animal diseases prevailing in the region.  To develop human resources for the field level veterinary services.  To co-ordinate and support national animal disease control and eradication  To support and facilitate the national veterinary regulatory services.  To participate actively in collaborative and coordinated research program in animal health and production in the region.  To support animal health and infertility camps in the region. 3. Organization Structure:
(S.V.O.)
Planning, Computing Reporting Monitoring, Evaluation Information Haematology
Pathology
Serology
Unit (V.T.)
Unit (V.T.)
Unit (V.O., J.V.T.)
Unit (V.O., J.V.T.)
Unit (V.O., J.V.T.)
Unit (V.O.,
Financial Unit
(Na.Su.)
Accountant
Annual Progress Report (2071/2072)
Activities
Progress
Blood protozoa identification EPG counts of parasites Skin scrapping examination Pathology
Clinical hematological examination Bio-Chemical examination Urine examination Post-Mortem Examination Isolation , identification and antibiogram of Bacteria Sample collection, Isolation and Identification of Fungus Virology
PPR Diagnosis by Pen site Test Serology
Examination for Mycoplasma Examination for salmonella Sample collection and examination
Sample collection and examination Disease surveillance and investigation
Regional Emergency disease investigation team (EDIT) mobilization Sub-clinical mastitis Investigation in Dairy animals Examination of antibiotic residues in meat and milk Identification, recording and reporting of regionally prioritized diseases
Livestock epidemic disease
Investigation
Collection and examination of samples
from cattle and buffaloes in infertility management camps Activities
Progress
Preparation of hematological profile of cattles in eastern region Zoonotic disease investigation
Examination for Brucella in animals Examination for Rabies (rapid test) in Examination for Tuberculosis in animals Number Staff Development
Training on Laboratory result collection Interaction programs
Technical workshop on livestock disease diagnosis (Regional level) Workshop on field sample collection and dispatch (District level) Participation in program planning, progress review and central level Laboratory Monitoring
Monitoring and evaluation of district level laboratories Publication of semi-annual epidemiological Report Publication of annual epidemiological Report Preparation and dispatch of Publication materials for Laboratory Technical Report Sending semi-annual epidemiological Purchase of Books and Journals Laboratory Management
Anti-rabies vaccination for staffs Health examination of staffs Lab animal management Sample collection for sero-monitoring of Times National PPR control program Sample collection for sero-monitoring of Times FMD control program Sample collection for sero-monitoring of Times Swine fever disease control program Sample collection for sero-monitoring of Times Ranikhet disease control program 4. Laboratory Services:
The routine laboratory works of RVL, Biratnagar, involve examination of fecal samples,
CMT and MWT tests of milk samples. Cultural examination of mastitis positive milk
samples are done to isolate and identify the bacteria responsible for this disease. Blood
samples are received here, particularly for Hb, PCV, TC, DLC tests, total protein and
blood protozoa identification. Serum samples are used to estimate Ca and P level in the
blood of animal. Similarly, serological test is done to screen brucella affected animal and
salmonella and mycoplasma affected poultry in this region. Hypersensitivity test
(tuberculin test) is done to isolate tuberculosis affected animal. Examination of skin
scraping and urine samples is frequently done in RVL, Biratnagar. In addition to this
different poultry diseases are to be diagnosed by performing rapid test and postmortem
methods.

4.1 Parasitological examination:
In this examination, both internal and external parasites are identified from the samples.
For internal parasites, fecal examination of Goat is done routinely. The fecal samples are
received mainly from farmers, DLSOs and also collected from field during investigation
programs. Most frequently the fecal examination is done by sedimentation and floatation
techniques to identify the gastro-intestinal parasites. However, in certain cases, Mc
Master Technique is followed to quantify the eggs per gram (EPG) in feces.
Details of Fecal examination in F/ Y 071/072 Location: Jhapa, Morang, Saptari Total Sample: 127 Positive Sample: 93(73.23%) Detail of parasites infestation, EPG:-400-500 In the fiscal year 2071/072, altogether 127 fecal samples of commercial goat from three
districts were received and examined. Among 127 samples, 93 samples (73.23%) were
positive. The result of fecal examination revealed that Hemoncus (47 %) is the most
prevalent parasitic infestation followed by Strongylus (35 %), Trichuris (13 %) and
others (5%).

4.2 Haematological examination:
Under haematological examination, TLC, TEC, DLC, PCV and Hb tests are performed in
this lab. Hb estimation is done by Sahli's haemoglobinometer, PCV by microhaematocrit
method, total count of RBC and WBC by Haemocytometer. Likewise, for DLC, blood
samples are stained with Giemsa.
Blood samples received from different districts of eastern region were examined for
blood parasites. A total number of 544 samples were examined for blood parasites. Out
of them, 265 samples were positive with dominance of Trypanosoma (243) followed by Babesia (13) and Theileria (9). Comparative chart of blood parasite (066/067 to 071/072) 066/067 067/068 068/069 069/070 070/071 071/072
4.3 Microbiological examination:
Altogether 1043 milk samples were registered in this laboratory in the fiscal year
2071/072.Out of them 469 samples were positive for CMT and MWT tests. The most
prevalent bacteria isolated from these positive milk samples were E. coli, staphylococcus,
Streptococcus, Klebsiella, Pseudomonas, Enterobacter etc.

M ilk s ample tes t for Mas titis Comparative c hart of CM T (067/ 68 to 071/72)
4.4 Pathological Examination:
Mostly postmortem examinations of dead birds and occasionally of dead animals are
done in the laboratory. During postmortem examination impression smears, swab, tissues
were collected for required tests. Altogether 432 dead birds were registered in the
laboratory. Diagnosis was done on the basis of PM examination and laboratory tests. Out of 432 samples, IBD had higher incidence followed by CRD and colibacillosis. PM examination of poultry Tentative diagnosis of Poultry diseases on the basis of P.M. examination & Lab tests – Total - 432 20, 5% 13, 3% 8, 2%
4.5 Biochemical examination:
Examination of urine and analysis of blood is routinely done to assess the different
conditions of urine and blood constituents. Serum samples are collected from farms,
animal health camps and sites of investigation program. Altogether 114 serum samples
were collected and analyzed during the fiscal year 071/072 for the estimation of Calcium,
Phosphorus, Total protein, Glucose, using specific kits. 36 Urine samples were tested by
using dipsticks (multisticks) as well as biochemical methods. Examination of urine was
done for specific gravity, PH, sugar, albumin, ketone bodies, urobilinogen etc. Mostly
Rothera's test and Robert's test were done to detect ketone bodies and protein
respectively.

4.6 Serological Examination:
Serological examination is done mainly for three diseases (Salmonellosis,
Mycoplasmosis and Brucellosis) in this laboratory. In fiscal year 2071/072 altogether 101
samples were tested for Pullorum disease by Plate agglutination method (PAT). 8
samples were found to be positive. Similarly, Rose Bengal plate test (RBPT) is done for
screening the Brucella positive animals. Total 149 (124 cattle and 25 goat) samples were
tested for detection of Brucella, out of them none of the samples were found to be
positive. Similarly, for Mycoplasma 185 samples were tested with 19 positive.

5. Epidemiological parameter of Eastern Development Region of Nepal:
Various epidemiological parameters of all the districts of eastern development region
were analyzed using different tools like table, graphs and pie-chart. Details of all those
analyzed information's are presented below.
Treatment Provided by DLSOs of EDR:
(Month-wise & District-wist), 2071/072
Month-wise no. of treated cases in EDR
No. of Treated Cases 2071/072


Situation of major diseases in EDR
In 1st, 2nd & 3rd quart of F/Y 2071/072
Informa tion about Reporting of Epidemiolog ica l Report from districts( 071 / 072) Detial of FMD outbreak in EDR
Detial of PPR Ooutbreak in EDR
Emergency quotas PPR Vaccine & Anti Rabies Vaccine Details
Anti Rabies Vaccine Distribution OIE PPR Vaccine Distributed Anti Rebies Vaccine Distibuted NVA Eastern Chapter Traning Center Duhabi Total distributed doses Available doeses in 071/072 Balance in 072/073 PPR Ranikhet, FMD, Swine Fever & H.S/B.Q. Vaccines Distribution under
National Programs
Dosages of Vaccine Distributed D. REGIONAL VETERINARY LABORATORY, JANAKPUR
(CENTRAL REGION)

1. Introduction:
Regional Veterinary Laboratory (RVL) of the central development region is situated in
Janakpur. It provides diagnostic services to all the 19 districts of central development
region of Nepal. Various disease diagnosed at the laboratory having several units i.e.
pathology, Parasitology, Microbiology, Hematology, Biochemistry, Sterilization.
Serological & Histopathological test results are obtained by dispatching the relevant
specimens to CVL, as these diagnostic facilities are not available in RVL Janakpur at
present. The Working areas of this RVL is all district of central region but mainly
focused on surrounding district like Dhanusha, Mohottory, Bara, Parsa, Rauthat and
Sinduli Which are situated around Janakpur. In this Central Region there are 3 zones and
19 districts geographically the region is divided in to 3 Eco-zones, Mountan, Hill &
Terai.

Mountain :-
This ecozones lies in the northern part of the region covering Rasua, Sindhupalchok of
Bagmati zone. Livestock rearing in the main occupation of the farmers in this region
Yak, Nak, Chauri, sheep & goat are being reared.

Hills :-
Kathmandu, Bhaktapur, Lalitpur, Dolkha, Makawanpur, Sindhuli are under this region.
Farmers follow mixed farming system and agro-based livestock industries are their main
occupation Cattle Buffalo, Pig, Poultry & Goat are being reared in this region poultry
farming are also popular among the farmers.

Terai :-
Dhanusha, Mohottary, Sarlahi, Bara, Parsa, Rauthat & Chitwan are under this terai
region. Through traditional system of livestock farming in followed in this region
poultry, dairy & piggery are being Commercialized especially in east west high way and
its vicinity.

2. Objectives:

• To provide prompt and efficient disease diagnostic services to the farmers of the • To investigate and diagnose the epidemics in the region. • To assist and support DLSOs in disease diagnosis and epidemic control. • To supervise and assist in diagnostic services to basic and primary laboratories situated in DLSOs of the region. • To collect, analyze and predict the animal diseases prevailing in the region. • To develop human resources for the field level veterinary services. • To co-ordinate and support national animal disease control and eradication • To support and facilitate the national veterinary regulatory services. • To participate actively in collaborative and coordinated research program in animal health and production in the region. • To support animal health and infertility camps in the region. 3. Organization Structure:
Table: Staff of RVL Janakpur
(At the end of F/Y 2071/072) Name of staff
Work of Responsible
Dr. Laxman Bhahadur Dhakal Mr. Ram Ashish Sah Dr. Laxmi Naryan das Mr. Anirudra Sah Microbiology, Serology Mr. Ram Adhar Tiwari Biochemistry, Help desk Mr. Ram Ratan Ray Yadav Mr. Ram Bahadur Chudhari Administration, Store Mr. Hari Narayan Balmpaki Mr. Kula Nand Jha Mr. Surndra Mishra Office assistant Mr. Dinesh Datta Office assistant Office Attendant Mr. Bhakti Nath Jha Computer Operator Computer Note -
S.V.O. :-
Senior Veterinary Officer Veterinary Officer Veterinary Technician Junior Veterinary Technician Table: Annual Progress Report (2071/072)
Programmers
Progress
1.
Laboratory Service
1.1
Parasitological Examination
E.P.G. Counts & Larva Culture Blood protozoa Identification 1.2
Pathological Examination
Clinical hematological Examination Postmortem Examination 1.3
Microbiological Examination
Bacterial Culture & Identification Antibiotic Sensitivity test Fungus Culture & Identification 1.4
Virological Examination
Virological Examination for Birds (Ranikhet & Nos. HA/HI Examination PPR Pen side Examination 1.5
Serological Examination
Micoplasma Identification Programmers
Progress
Sallmonella Identification Elisa Test (PPR) Sample Collation & dispatch 1.8
Disease Investigation & Surveillance Program
me
Clinical Sub Mastitis Test & repotting Antibiotic Resistant Bacteria Serve & Scaring Antibiotic Residue Test milk & meat Regional Emergency district investigation team Times Investigation of Epidemic Disease 1.9
Identification of Zoonosis Disease
Avian Influenza examination Brucella examination Rabies examination Vaccine Bank management
Regional PPR Vaccine bank management Regional Rabies Vaccine bank management 3
3.1
Seromonitoring for PPR sample collection & Times 3.4
Seromonitoring for Ranikhat sample collection & Times 4
Follow up District Level Labortory 5
Veterinary disease investigation workshop Planning, Budget & Progress report Workshop 7
Publication of Half yearly Epidemiological Times Publication of Yearly Epidemiological Bulletin Preparation of Records for yearly publication 8
Purchasing of books & Journals
9
Lab animal management
10
Contract Service
Purchasing of machinery equipment Computer operator, Sweeper & mali 11
Office Structure Management

4. Laboratory Service:

The routine Laboratory Works of RVL Janakpur , mainly involves examination of faecal
Sample, Postmortem Examination, Sero-Surveillance, CMT test of milk samples and
culture examination of mastitis. Positive milk samples are carried to isolate and identify
the bacteria responsible for this diseases. Blood samples brought here from DLSOs
particularly for HB, PCV, TC, DLC, Total protein and blood protozoa identification.
Examination of skin scraping & Urine test was frequently conducted in RVL Janakpur.
Antibiotic sensitivity test are in regular basis after bacterial culture of positive sample.

4.1 Parasitological Examination:

Parasitological examination (Internal & external), fecal examination of different animals
has been done routinely. The fecal samples are received mainly from farmers referred by
DLSOs and also collected from Dhanusha, Mahottary, Sarlahi, Sinnduli, Rauthat and
Bara, field area during surveillance and investigation programme. For the gastro
intestinal parasites, Mc'master technique is followed to quantity the eggs per gram (EPG)
in faeces.
In the F/Y 2071/72 total 208 faecal samples from different species of animals were
received & examined. Among these samples 106 samples (52%) were positive. The
results of faecal test revealed that Liver fluke (25%), Paramphistomum (48%), Trichuris
(11%), Strongylus (9%), Mixed & Other parasites (7%) were prevalent in those samples.
Table: Figures of faecal sample examined and types of parasites identified
Strongylus
Trichuris
4.2 Haematological examination:
Haematological examination TLC, TEC, DLC, PCV, & HB test are done in this
laboratory. Total count of RBC, WBC by Haemocytometer for DLC blood sample were
stained with Giemsa blood samples received from different district of central region.
Total 305 Blood sample were examined for different blood parameters as well as for
blood parasites. Among them 224 samples were found negative for any blood parasites &
rest 81 were found positive for different blood parasites.
Table: Details of blood sample examined
Anaplasma
Babesia Theileria
Negative
4.3 Pathological Examination:
The pathological examination includes mostly post mortem examination of the dead birds
received from commercial poultry farms. About 99% cases were bought from Janakpur
municipality area & remaining of samples from rural area of the Dhanusha district for
pathological examination. A total 250 cases of postmortem examination were presented
during the F/Y 2071/072 all the carcasses received was avian. No cases of large & small
ruminants and other species of animals were received. The status of poultry disease in the
area is shown in Table.
Table: Distribution of various poultry diseases diagnosed in the laboratory
Total Cases
Tentative Diagnosis
Infectious Bursal Disease New Castle Disease Litchy Heart Disease
4.4 Microbiological Examination:
Various bacterial species were identified in cattle and buffalo milk. A total of 328 Cattle
& Buffalo milk sample collected from the following district & samples were tested for
the presence of Bacteria in milk. The most prevent bacteria isolated in the cow and
buffalo milk was Staphylococcus sp., Streptococcus sp. and E. coli.
Table: Bacteria isolated for milk samples of cattle and buffalo in different districts
Tested No. of
District
Staplylococcus, Streptococcus & E. coli Staplylococcus, Streptococcus & E. coli Staplylococcus, Streptococcus & E. coli Staplylococcus, Streptococcus & E. coli Staplylococcus, Streptococcus & E. coli Staplylococcus, Streptococcus & E. coli Table: Major antibiotics sensitive to bacterial isolates
Antibiotics Used
Percent Efficacy
4.5 Serological Examination:
Pate agglutination test for three diseases were conducted in the laboratory. All together
108 serum sample were tested for the presence of Mycoplasma sp., among them 29 were
+ve & rest were found negative. Likewise, 147 samples were tested for Salmonella with
64 +ve and for Brucella also 143 samples tested with 17 positive results.
4.6 Biochemical Examination:

Table: Values of serum calcium and phosphorus detected in various livestock
Phosphors
Phosphors
gm/100ml
gm/100ml
gm/100ml
gm/100ml
of Sample
Normal Value
Normal Result
Normal Result
5. Sample sent to CVL for further Investigation in F/Y 2071/072:
Regional Veterinary Laboratory, Janakpur is not well equipped with the modern
equipments. The samples are also sent to CVL for diagnosis of the disease &
reconfirmation of the different types of disease diagnosed. FMD suspected samples were
also sent to National FMD and TADS Control Laboratory, Budhanilkanth, Kathmandu.
During the fiscal year 2071/072 altogether 750 samples were dispatched to CVL and
National FMD and TADS Control Laboratory for reconfirmation and diagnosis.
Table: Types of samples sent to Central Veterinary Laboratory
Types of Samples
Table: National PPR Programme (Sero-Surveillance of F/Y 2071/072)
Name of DLSOs
Serum to be collected (No.)
Surveillance in Central Development Region, RVL Janakpur
Types of sample
collected
District
E. REGIONAL VETERINARY LABORATORY, POKHARA
(WESTERN REGION)
1. Introduction:
Western Development Region (WDR) is situated between 82º 30' to 85º 15' east
longitude and from 27º 15' to 29º 30' north latitude. It occupies about 20% (29355 Sq.
Km.) of total areas of Nepal. The region shares boundaries with Uttar Pradesh of India in
the south and Tibet of China in the north. The region is bulging between Central and
Mid-western development regions of Nepal in the east and west respectively.
Geographically, WDR is divided into the following three main domains:
Himalayan region
Himalayan region is located in the northern part of the WDR, covering Mustang, Manang
and upper belt of Gorkha districts. Yak/Nak, sheep, alpine goats (Chyangra) and mule
rearing formed the way of life of the people in this region.
Hilly region
Hilly region lies in between the Himalayan and Terai regions. This region
comprised of Arghakhanchi, Gulmi, Palpa, Shyanga, Kaski, Tanahu, Lamjung, and lower
belt of Gorkha, Parwat, Baglung and Myagdi districts. People of divergent ethnic groups,
casts and cultures share their common way of living. Agro-based livestock industry in
this region is the main source of income of the people. Poultry farming, goat rearing and
dairy industries are becoming familiar near the cities/towns and in the areas where
market is accessible.
Terai region
Terai region covers Nawalparasi, Rupandehi and Kapilbastu districts. This plain
extends from east to west of the region and stretched from 15 to 40 Kilometers in width.
Sediments and silt are main constituents of soil deposited by rivers making it more fertile
and this belt supplies the food and fibers to other regions of the country. Compared to
mountains and hilly regions, this region has relatively better infrastructure and market
accessibility. People of this region are motivated to adopt livestock farming in
commercial scale.
The population of livestock in the region is very high as compared to their production.
Though many factors are contributory, the health of animal plays a vital role to increase
the production and productivity of animal. Every year, several diseases and parasitic
problems attribute a considerable amount of economic loss to livestock rearing farmers of
the country warranting switching on a massive disease control program. Major
economically important diseases of cattle and buffaloes are Foot and mouth disease
(FMD), Haemorrhagic septicemia (HS), Helminthiosis, infertility, mastitis and blood
protozoan diseases whereas PPR, Gastrointestinal nematodosis and Clostridial diseases
in sheep and goats. Swine fever and FMD in pigs and Avian Influenza, New Castle
disease (ND), Infectious bursal disease (IBD), Coccidiosis, Hemorrhegic Enteritis and
Mycotoxicosis in chickens are major disease problems.
2. Objectives:
• To provide accessible, timely and accurate diagnostic services to the livestock and poultry farmers and to veterinarians, veterinary technicians and their owners in the region. • To conduct diagnostic examinations, record results, report information, and assist in the interpretation of results to submitting DLSOs, Veterinarians, and veterinary technicians. • To investigate the animal disease epidemics in the region and assist, advice and support DLSOs to control them. • To prepare epidemiological profile of livestock and poultry diseases and maintain and disseminate the regional epidemiological information database on animal health in the regional as well as in the national networks. • To investigate relatively important livestock diseases in the region and formulate control measures for the same with wider consultation to the experts. • To monitor and report the incidence and threat of animal diseases, as well as diseases that are transmissible form animal to humans. • To supervise and assist in diagnostic services to basic and primary laboratories based at DLSO's of the region. • To conduct and support the laboratory and animal health related training programs for the Para vets in the region. • To coordinate national disease control and eradication programs in the region. Table : Annual Program and summary of achievements for
fiscal year 2071/72
Annual Targets
Programmes and
Activities
Weightage Progress age (%)
Capital Expenditure
Construction of
Remaining Compound Purchase of Fridge Running Expenditure
Laboratory Services
Examinations EPG Count and Larva Culture Examination of Blood Parasites Pathological Examinations Clinical Hematological Tests Calcium, Phosphorus, T. 1.2.2 Protein, Glucose, 1.2.3 Urine Test Annual Targets
Programmes and
Activities
Weightage Progress age (%)
1.2.4 Postmortum Examination Examinations Isolation and Identification of Bacteria Isolation and Identification of Fungus Virological Tests (Ranikhet,Gumboro,AIV) PPR testing using penside Serological Tests Mycoplasma tests in Poultry using PAT Salmonella tests in Poultry using PAT Disease Investigation and Surveillance Program
Management of Regional
EDIT Investigation of Epidemic
Laboratory Diagnosis of
Zoonotic Diseases
Brucella Diagnosis using
Rabies Rapid Test Tuberculosis Testing using Tuberculin Monitoring and Supervision
Follow up tour of district Laboratories management Workshops and seminars
Investigation Interaction Publication Program
Epidemiologic Bulletin publication Annual Epidemiological Bulletin Publication Material Preparation for Annual Technical Report Purchase of Journal
Laboratory
Management
Civil Servent Use in Anti Porson Annual Targets
Programmes and
Activities
Weightage Progress age (%)
Rabies Vaccine Civil Servent Health Lab Animal Management Month Contract Service
Machinery Tools and
Computer Management
Computer
per,Driver and Lab Equipments Repair Management etc. P.P.R. Sero-Monitoring F.M.D Sero-Monitoring Swine Fever Sero- Monitoring Ranikhet Sero-
3. Laboratory Services:

3.1 Parasitological Examination:
Parasitological Examination of Blood Protozoa in the fiscal year 2071/072 Blood sample
Staining Blood smears. A total of 106 blood samples from animals were examined for
different Animal.

3.2 Microbiological Examinations:
Microbiological examinations include the isolation and identification of bacteria and
fungi from the pathological samples received in the laboratory. Bacteriological culture,
Fungus Culture and antibiotic sensitivity tests were performed of the samples received
for microbiological investigation. During 2071/72 a total of 521 samples were examined
in microbiology unit of the laboratory.
Table: Different Bacterial isolates of Mastitis
Name of the isolates
No. of Cases
Percentage of cases
Staphylococcus sps. Streptococus sps. All the organisms shown in the culture were subjected for the antibiotic sensitivity test and gave the following result: 3.3 Pathological Examinations:
Pathological examinations mostly consisted of necropsy examination of carcasses
presented in the laboratory where poultry dominates all. In the pathology unit, the cause
of death of chickens presented was generally drawn on the basis of both the post mortem
lesions observed and laboratory investigation of samples collected during necropsy
examinations.

Table: Diseases of chickens diagnosed on the FY 2071/72
Diseases
Percentage
Infectious Bursal Disease New Castle Disease Others (Enteritis, Hepatitis,etc) Total cases (n) = 2537 Maximum cases are mixed infections

It can be seen during the FY 2071/72, Colibacillosis was the most prevalent poultry
disease followed by Mycotoxicity. There was increase in the cases of Colibacillosis,
which might be due to the increased number of farming and poor management condition
of the farming system.
4.4 Serological examinations:
Serological examinations mainly consisted of plate agglutination test of chicken serum to
etect antibody against Mycoplasma gallisepticum and Salmonella pullorum organisms.
Similarly, serum samples from cattle, buffalo, sheep, goats and dogs were tested for
brucella antibodies using Rose Bengal Plate Agglutination Test (RBPT). During the
fiscal year 2071/72, the serum samples tested and their results are presented as follows:
Table: Serological Test Result in RVL in FY 2071/72
Percentage
Tested For
of Positives
Table: Virological test results of samples of different species
Percentage
Number of
Test Method
Tested For
Positives

3.5 Hematological Examinations:
Hematological unit of the laboratory is well equipped to determine a range of
hematological parameters such as Total Erythrocyte Count (TEC) and Total Leukocyte
Count (TLC), Differential Leucocytes Counts (DLC), Erythrocyte Sedimentation Rate
(ESR), determination of hemoglobin (HB) and Packed Cell Volume (PCV) and staining
of blood smears for blood protozoa and bacteria. A total of 106 blood samples from
animals were examined for different hematological parameters.

3.6 Biochemical examinations:
Biochemical examinations included biochemistry of serum and routine and microscopic
examination of urine. Multistick strip was used for routine urine analysis. Microscopic
examination of urine was done after centrifugation of the urine samples.

3.7 Sample Collection and Dispatch:
During 2071/72, serum samples, brain and tissue samples of different animal species and
poultry were collected from the disease investigation sites. A total of 474 various samples
were dispatched to Central Veterinary Laboratory, Kathmandu and National FMD and
TADs laboratory, Kathmandu for confirmatory disease diagnosis.

3.8 Avian Influenza Surveillance (Laboratory Surveillance)
The annual avian influenza laboratory surveillance programme was continuing in the
laboratory and it was successful in the detection of flu A cases. During this fiscal year
071/72 all together 17 flu A cases were identified out of 2537 cases registered for the
disease diagnosis. Four out of 17 flu A case were confirmed as H9 by CVL.
F. REGIONAL VETERINARY LABORATORY, SURKHET
(MIDWESTERN REGION)

1. Introduction:
Regional Veterinary Laboratory (RVL) of the Mid-western Development Region (MDR)
is situated in Birendranagar Municipality-7, Surkhet and was established in FY
1988/1989 AD. It provides laboratory diagnostic services to all the 15 districts of MDR.
In this region, there are 3 zones (Rapti, Bhari and Karnali) and 15 districts.
Geographically, the region is divided into 3 eco-zones (high hills, mid hills, and terai). To
provide the diagnostic livestock services smoothly, three districts (Banke, Dang and
Jumla) have established basic laboratories. They perform parasitological examinations,
microbiological culture and antibiotic sensitivity test. The mission of the RVL, Surkhet is
to promote the health of livestock, poultry and ensure safe animal products for consumer
by assisting DLSO's offices, veterinarians, clients, and others who are responsible for
animal health in detection and prevention of animal diseases.
2. Objectives:
• To provide prompt and efficient disease diagnostic services to the farmers of the • To investigate and diagnose the epidemics in the region. • To assist and support DLSOs in disease diagnosis and epidemic control. • To supervise and assist in diagnostic services to basic and primary laboratories situated in DLSOs of the region. • To collect, analyze and predict the animal diseases prevailing in the region. • To develop human resources for the field level veterinary services. • To co-ordinate and support national animal disease control and eradication • To support and facilitate the national veterinary regulatory services. • To participate actively in collaborative and coordinated research program in animal health and production in the region. • To support animal health and infertility camps in the region. 3. Organization Structure:

Table: List of staff of Regional Veterinary Laboratory, Surkhet (FY 2071/072).
Designation
Number Fulfilled
Senior Veterinary Veterinary Officer Assistant Animal Health Technician Table: Annual work program and summary of achievements (FY 2071/072)
Progress
Activities
Target Progress
Laboratory services Parasitological Examination, identification and recording EPG Count, Larvae Culture Identification and Treatment Blood Parasite Examination Pathological Examination Clinical Hematological Examination Biochemical Examination Urine Examination Microbiological Examination Isolation, Identification and Recording of Bacteria, Fungus and Sensitivity Virological Tests Viral disease Examination in Poultry HA, HI Examination PPR testing using Pen site Test Serological Examination Mycoplasma Examination in Poultry Salmonella Examination in poultry Sample collection and dispatch Sample collection and dispatch for further examination in other Laboratories Epidemic Investigation Subclinical mastitis investigation and reporting in Cattle and Buffalo Progress
Activities
Target Progress
Antibiotic Resistance Bacteria Surveillance and shearing Antibiotic Residue examination (Milk, Regional label emergency Disease investigation team management Epidemic investigation Disease investigation, filing, reporting in regional label Zoonotic Disease Investigation laboratory Program Avian influenza examination Brucella examination Rabies examination Tuberculosis examination Vaccine Bank Management Vaccine Bank Management in Regional Rabies Vaccine Bank Management Sample collection and dispatch of PPR for sero monitoring Sample collection and dispatch of FMD for sero monitoring Sample collection and dispatch of Swine fever for sero monitoring Sample collection and dispatch of ND for Sero monitoring Monitoring and supervision Supervision and Monitoring of District Work Shop program Animal Disease Investigation interaction workshop District label Laboratories training program (1 Days) Publication program Half yearly epidemiological bulletin publication (preparation). Yearly epidemiological bulletin publication (preparation). Annual Technical Bulletin Publication Purchase of Books and Journal Management of Lab Animal Contract Service Laboratory Equipment Preservation Computer Operator, Driver, Others 4. Laboratory Services:
4.1 Biochemistry unit:
Biochemistry unit analyzes mainly serum for the estimation of calcium, phosphorus,
Glucose, Bilirubin and total proteins at present of cattle, buffalo, goats and sheep. This
unit performs the urine tests for the estimation of albumin, bilirubin, urobilinogen,
specific gravity and blood by using dipstick test kit.
Table: Biochemical test result of 2071/72.
No. Below
No. Above
No. of Normal
Biochemical Test
Normal Level
Normal Level
4.2 Hematology unit:
The Hematology unit is providing routine hematological parameters of all the animals
and poultry. Hematological parameters include Total Erythrocyte Count (TEC) and Total
Leukocyte Count (TLC), Differential Leukocyte Count (DLC), determination of
hemoglobin (Hb) and Packed Cell Volume (PCV), and staining of blood smears for blood
protozoa. Anaplasma and Babesia species was prevalent in blood samples collected from
improved breed of cattle from Bardiya and Surkhet districts.
Table: Results of Plate Agglutination Test
S. Name of Agglutination Test
No of sample No of Sample Percentage Rema
Positive
Salmonella Agglutination Test Mycoplasma Agglutination Test Rose Bengal Agglutination Test(Brucella) PPR Penside test Table: Results of California Mastitis Test
Positive
Percentage
Table: Rapid Test Kit Result during 2071/072
No of sample No of Sample Percentage
Name of Disease
Positive
Antibiotic Residue Tetracycline 100ppm+ 4.3 Parasitological examination:
Parasitological unit examines fecal samples of various species using different methods as
direct smear, sedimentation, and floatation methods. Parasitological unit not only identify
the parasites but also quantify the parasitic burden of the nematode, trematode and
cestodes by McMaster method. In this fiscal year 2071/072, total 577 faecal samples
were examined and 258 were found positive for various internal parasites. Among them
Fasciola, Coccidia, Strongylus, Paramphistomum were found major internal parasites
identified. Result of faecal examination is presented in table and figure below.

Table: Monthly Fecal Examination Result during 2071/072

Parasite
Paramphistomum 1 Sample Total tested Figure: Result of fecal examination during 2071/072
4.4 Microbiological examination
Microbiological unit examines diversified samples like, tissues, blood, aspirated fluids
and tissues etc. Both aerobic and anaerobic culture facilities are available. The unit also
performs identification of the bacterial and fungal organisms. The Microbiology unit also
performs antibiotic susceptibility test and advice for the appropriate antibiotic for the
treatments. The bacteria isolated from milk samples were E. coli, Staphylococcus spp and
streptococcus spp. Ciprofloxacin, gentamicin and enrofloxacin were highly sensitive
against the isolates while tetracycline, amoxicillin and cloxacillin were found resistant.
Table: Isolated Micro-organism during 2071/072
Name of organism
No of Isolation
Streptococcus sp. Staphylococcus sp. Name of organism
No of Isolation
Pasteurella Multocida Campylobacter sp. Microbiological examinations include the isolation and identification of organisms from the pathological samples. Bacteriological culture and antibiotic sensitivity test is conducted for microbiological investigation. The samples were milk, urine, tissue, skin scrapings etc. The organisms shown in the culture were subjected to sensitivity test. Antibiotic sensitivity test conducted at RVL Surkhet, during 2071/072 the test result is given below. Figure: Antibiotic sensitivity test result of RVL, Surkhet (FY 2071/072)
4.5 Pathological examination:
The pathological examination includes mostly post mortem examination of poultry
received from commercial poultry farms. Most of the cases were brought from
Birendranagar Municipality and surrounding VDCs; Utterganga , Latikoili ,Chhinchu,
Jurbutta and some VDCs of Bardiya and Dang District for pathological examination. The
status of poultry diseases according to post-mortem examination is shown in table and
figure below.
Table: Monthly Pathological Examination Result during 2071/072
Name of Disease
Newcastle Disease Figure: Pathological Examination Result during 2071/072

4.6 National PPR Program:
Under National PPR Control program, Directorate of Animal Health had provided
430000 dose of PPR vaccine for 11 districts of this mid western region. RVL Surkhet
supported the program by sero-monitoring. District-wise collection of serum sample is
presented in the table below.
Table: District wise collection of serum sample.
District
Vaccination
Serum collection
Progress
Table: PPR Seromonitoring with ELISA Test
Antibody
Percentage
Positive
46.7 % Total sero Disease investigation
4.7 Swine Fever Sero-monitoring Program:
The total number of swine fever Vaccines distributed from Directorate of Animal Health
was 60000 dose under national swine fever vaccine program for 3 districts for this
region. RVL, Surkhet supported the program by sero-monitoring. The table below shows
district wise collection of serum sample.
Table: District wise collection of serum samples
Serum collection
District
Vaccination
Progress

4.8 FMD Sero-monitoring Program:
The total number of FMD Vaccines distributed from Directorate of Animal Health was
40000 doses under FMD vaccine program for 2 districts for this region. RVL, Surkhet
supported the program by sero-monitoring. The table below shows district wise
collection of serum sample.
Table: District wise collection of serum samples
Serum collection
District
Vaccination
Progress

4.9 Ranikhet Disease Sero-monitoring Program
The total number of ND Vaccines of poultry distributed from Directorate of Animal
Health was 20000 doses under national swine fever vaccine program for 2 districts for
this region. RVL, Surkhet supported the program by sero-monitoring. The table below
shows details.
Table: District wise collection of serum samples
District
Vaccination
Serum collection
Progress

5. Epidemic Investigation:
Various disease outbreaks of animal and poultry were investigated during F/Y 2071/72.
Whenever request for investigation of an outbreak was received from the district to the
RVL, a veterinarian or a technician or a team of technicians with necessary sampling kit
visited to the site of epidemic, collected epidemiological information and appropriate
pathological samples. In the laboratory, pathological samples collected from the field
were processed to find out the etiology of the outbreak. Epidemiological information
gathered from the site of the outbreak was used to decide the test to be performed in the
laboratory and to assist in the confirmation of disease diagnosis. Samples, not possible to
process in this laboratory were referred to CVL, Kathmandu.
PPR and FMD were reported from Surkhet. In Jumla district PPR were found to be
affected as well as Enterotoxaemia and massive parasitic infestation. In Banke, Bardiya,
Dang and Surkhet IBD, ND, Colibacilosis and Coccidiosis were reported frequently in
poultry. Most of outbreaks investigated were confirmed by laboratory while few
confirmations were based of clinical signs and postmortem findings.
F. REGIONAL VETERINARY LABORATORY, DHANGADHI
(FAR WESTERN REGION)

1. Introduction:
Regional Veterinary Laboratory is situated in Dhangadhi municipality of far western
Nepal. This laboratory is established as the reference laboratory of the region with its
service area covering the nine district & two zones. Regional Veterinary Laboratory,
Dhangadhi was established in 2049/050.
Geographically the region is divided into three parts namely, Mountains, Hills and Terai.
The mountainous districts of the region comprises of Bajura, Bhajhang, and Darchula,
Likewise, hilly districts of the region are Baitadi, Dadeldhura, Acchamm, & Doti. Kailali
and Kanchanpur are the Terrain districts of the Far western region. All districts have a
high potentiality for the development of livestock industry. Almost all kinds of livestock
viz cattle, buffalo, sheep, goat, pig, poultry, duck, etc. are reared in the region. On the
other Goat pig, cattle and poultry farming is inclined towards commercialization, but
particularly poultry farming is rapidly gaining popularity among the farming
communities.
There are a number of infectious diseases which are constrain to the development of the
livestock sector of the region, apart from management, nutritional and other factors, the
major being the FMD, PPR, Swine Fever, HS protozoan disease. Now Layer and broiler
are reared in commercial scale in Kailali and Kanchanpur districts. Recently some
farmers of Dadeldhura,
Baitadi and Doti are showing interest in poultry farming, and they have started to keep poultry in number of few hundreds. A few economically important diseases of poultry in this region are New Castle, Infectious Bursal Disease, Coccidiosis, Chronic Respiratory Disease (CRD), Inclusion Body Hepatitis (IBH), Leechi Heart Disease, Collibacillosis, etc. However, the disease diagnosis is still not based fully on the laboratory findings. It is most of the times based on the history, statements put forward by the farmers, and the clinical findings of the animal on examination. 2. Objectives:
• To provide prompt and efficient disease diagnostic services to the farmers of the • To investigate and diagnose the epidemics in the region. • To assist and support DLSOs in disease diagnosis and epidemic control. • To supervise and assist in diagnostic services to basic and primary laboratories situated in DLSOs of the region. • To collect, analyze and predict the animal diseases prevailing in the region. • To develop human resources for the field level veterinary services. • To co-ordinate and support national animal disease control and eradication • To support and facilitate the national veterinary regulatory services. • To participate actively in collaborative and coordinated research program in animal health and production in the region. • To support animal health and infertility camps in the region. 3. Organization Structure:

Table: Staffing of RVL Dhangadi (At the end of F/Y 2071/072)
Type of post
Class/post
number fulfilled
Table: Annual program and Summary of Achievement in F/Y 071/072
Programs and Activities
Progress
Progress
Percentage
Furniture and fixer Laboratory Service programme- Parasitological Examination Identification of parasites and EPG Identification of blood protozoa Skin scraping Test Clinical hematological examination Biochemical examination Pathological examination Postmortem examination Microbiological culture & Identification Antibiotic sensitivity Test fungus culture& Identification Viral diseases examination of Birds(ND/IBD/AI) examination Salmonellosis examination (PAT) Sample collectin test & Dispatch Sub clinical mastitis investigation in Investigation of Kumri in goat Regional level Investigation of Emergency Diseases Epidemic investigation Programs and Activities
Progress
Progress
Percentage
Zoonotic disease investigation Vacine Bank Management PPR & PPR Seromonetaring & Sample FMD Seromonetaring & Sample Swine fever Seromonetaring & Ranikhet Seromonetaring & Sample Inspection and Supervision of District Time Techinical Interacton of animal Distric leval lab Technique training Participation in budget, program,& Publication Programme: Hajf Yearly Epidemiological bulletin Yearly Epidemiological Bulletin Annual Technical Book material Anti Rabise vaccine for staff Health checkup for stff Management of Lab.animal Protection of Lab.equipments & Office assistant, computer operater & Driver for contact 4. Laboratory Services:
4.1 Parasitological Examination:
Altogether196 samples were tested for different parasitic conditions of livestock
populations. Samples for examination mainly consisted of the regular fecal samples
submitted to the district livestock service office, Kailali. Apart from this samples were
also collected from field during epidemic disease investigation and the investigation
programme in our set annual programme. The most common helminthes parasite
identified during faecal examination was Fasciola followed by other internal parasites of
nematode group, viz. Strongylus, Trichuris, Coccidiosis,, etc. It is found that most of the
fecal samples examined at the RVL are positive for one or the other internal parasites.
Out of 196 samples tested 48 (24.5%) were positive. A total of 196 samples were tested
for EPG count to see the prevalence of internal parasite in Goat. In EPG counting the
prevalence rate was 24.5% in goats. Stronyles. Strongyloids, Trichuris and Monezia
were the most frequently observed parasites in those samples.

4.2 Serology:
A total of 560 serological tests of different types were performed against the target of 400
during the fiscal year 2071/72 Most of the serum samples collected were from goats for
various diagnostic tests Brucellosis and salmonella for poultry disease conditions
responsible for causing abortion in these animals, some of the samples were forwarded to
CVL.

Serum samples collected from bovine, caprine & poultry population were mainly
obtained from Baitadi, Kailali, Kanchanpur, Doti, Dadeldhura and Darchula. Regular
screening test of Brucellosis and Salmonellosis was performed using Rose Bengal Plate
Agglutination Test & Salmonella PAT.
Table: Result of Serological examination performed at the RVL, Dhangadhi
Positive
Disease Name
Sample Tested

4.3 Hematology:
A total of 205 samples were examined for Haematology at the RVL,Dhangadhi included
TLC, DLC, PCV, Hb examination for presence of blood parasites. Blood samples were
mainly collected from livestock during outbreak of epidemic and cases referred by the
DLSO"s.
On our examination of hematological parameters of parasitic infestation animals they
invariably showed low Hb concentration. Hb value in these animals ranged from low of 8
to a high of 18g./dl. & PCV ranges from16-42dl of blood. Similarly, 159 blood samples
were examined for blood protozoan and were negative for Babesia sp.

4.4 Pathology:
This unit of the laboratory mostly receives poultry& pig carcass for necropsy study and
disease diagnosis. However, dead bodies of other animal species are also received
occasionally, especially during disease outbreak and in cases of veterolegal importance.
Out of the total of 487 postmortems performed during the fiscal year 071/072 only 3
cases of pigs from Kailali, Dadeldhura, Kanchanpur and the rest were of poultry species.
Since most of the pathological samples comprised of poultry, it is important to present the major diseases diagnosed, based on findings of postmortem lesions. The common diseases that were diagnosed through postmortem are Cocccidiosis, IBD, IBH, Colibacilosis, CRD, Mycotoxicosis, Asitis, Gout & Fatty liver syndrome etc. Table: Pathological conditions diagnosed during postmortem examination
Percentage
Infectious Bursal Disease Chronic Respiratory Disease Newcastle Disease Sum Total

4.5 Microbiology:
The samples received for microbiological examination at the Regional Veterinary
Laboratory, Dhangadhi constitutes of milk, nasal swab, vaginal swab, and swab from
visceral organs like liver, lungs, intestine etc. of various animal species. The media used
for microbiological culture were Nutrient agar, Mcconkey agar, Blood agar, and
Saboroud Dextrose agar. Bacteria and fungi were identified on the basis of colony
characteristics, Gram's staining property and the structure of the organism as seen under
the microscope. Due to limitation of the facility in the laboratory biochemical tests for
identification of bacteria could not be performed. The Result of microbiological
examination is presented in the table as below.
Table: Microorganisms isolated from different samples during the F/Y 2071/72
Type of sample Number
Major bacteria identified
Strep., Staph., Cory. Strep., Staph., Cory., Strep., Staph., Bacillus E. coli, Strep., Staph., Aspergellus, Penecilium, E. coli, Strep., Staph., Coryne E. coli, Strep., Staph., The milk samples positive for California mastitis were tested for antibiotic sensitivity test following preliminary culture in order to choose right antibiotic for the treatment of mastitis. The result of antibiotic showed that Enrofloxican was more sensitive that other antibiotics. 4.6 Biochemistry:
Altogether 154 samples were collected & analyzed in this fiscal year for estimation
calcium, phosphorus, total protein & glucose. Biochemical examination of serum was
performed with the serum sample of buffaloes suffering from Khari disease mainly for
estimation of serum calcium & phosphorus level. However, the Calcium content of the
serum was recorded from a low of 7 to a high of 16mg/dl.
5. Investigation programs:
5.1 Kumri (Seteria Spp.) in Goats:
A part from above mentioned activities this laboratory has performed a Kumri in Goat
investigation program in Kailali & Kanchanpur district. The main objective of study was
to see the prevalence of Kumri in goat, especially in Hilly area & Tarai area of Kailali &
Kanchanpur district. Active surveillance was done through questionnaire, serum and
feacal samples were taken from infected goats.
This investigation programs was conducted in sites of Kailali district namely,
Ramsikharjhala & Mohanyal VDC. In this study a total of 180 house hold were
interviewed and sampled, there were 2311 goats in that area .Summary of investigation
and finding are given below.
Table: Prevalence of Kumri in goats in Kailali district
Total goats
Prevalence%
/Paralysis
Similarly, an investigation program was conducted in sites of Kanchanpur district namely Krishnapur VDC. A total of 86 house hold were having 516 goats were involved in this investigation program. Summary of investigation and finding are given below. Table: Prevalence of Kumri in goats in Kanchanpur district
Total goats
Prevalence%
/Paralysis

Overall Findings: A overall prevalence of 14.53 % of Kumri (Setaria spp.) was recorded
in Kailali and Kanchanpur districts during the month of Bhadra to Mangsir.

5.2 Sub clinical mastitis:
This program was conducted to see the prevalence of sub clinical mastitis in milking cow
& Buffalo of Kailali & Kanchanpur districts. So fresh milk samples were taken from
farmers directly and tested by SLST reagent immediately.

Investigation Site
Kailali (Masuriya Chaumala,& Darakh)
Kanchanpur (Suda VDC ,Daizzi VDC &,Tribhubanbasti, )
Objectives - To see the prevalence of sub clinical Mastitis in cows of Kailali &
Kanchanpur

Method–
Active Surveillance & Questionnaire. Milk sample were directly taken from
farmers, Test Method: Through SLST Reagent(Coagulation)
Table: Summary of Sub clinical mastitis Investigation in Kailali and Kanchanpur
Buffalo Milk
Table: Detail Report of Sub clinical mastitis in Kailali and Kanchanpur
.Cow milk sample Buffalo milk sample

Source: http://www.cvl.gov.np/uploads/files/5324820354.pdf

chemoemboli.ru2

Cardiovasc Intervent Radiol (2008) 31:735–744 Transcatheter Arterial Chemoembolization for AdvancedHepatocellular Carcinoma with Inferior Vena Cava and RightAtrial Tumors M. C. Chern Æ V. P. Chuang Æ T. Cheng ÆZ. H. Lin Æ Y. M. Lin Received: 24 September 2007 / Accepted: 17 March 2008 / Published online: 22 April 2008Ó Springer Science+Business Media, LLC 2008

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