Nepal Elephant TB Control and Mgt Action Plan.pdf - Elephant Care ...

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Nepal Elephant Tuberculosis Control andManagement Action Plan (2011-2015)Respiratory Sample Collecon ProceduresThe Nepal Elephant TB Control and ManagementAcon Plan has established collaboraons withthe Nepal An-tuberculosis Associaon, theCenter for Molecular Dynamics-Nepal (CMDN),and the experts in this field to improve methodsto idenfy TB bacterial shedding in elephants.The following procedures may change as newknowledge is gained.Loop-mediated isothermal amplificaon (LAMP)is a molecular technique that has been used todiagnose TB in humans in Nepal (Pandey 2008).It is accurate in humans and may prove to be aneffecve diagnosc tool for elephants but shouldbe performed in conjuncon with culture andPCR. Collect samples in the a ernoon a er theelephant has been acve and moving around.Collecon of trunk wash or liquid nasal dripsample for culture, PCR, and LAMP1. Wear an N95 mask and exam gloves.2. Pre-label a plasc 50 ml screw top centrifugetube with the elephant’s name, project ID#,facility locaon, and date of collecon. Thissample will be for culture.3. Pre-label a 4.5 buffer-filled nunc tube for DNAextracon, PCR, and LAMP.3. Collect standard trunk wash sample or 10ml of a nasal drip sample in a shallow largediameter plasc tub or a large jar.4. Transfer the sample into a plasc 50-mlscrew top centrifuge tube by pouring or use adisposable syringe or pipe!e. If the sample isgrossly contaminated, pour through a gauzesponge.5. Dip a clean sterile swab into the sample andplace the swab in the buffer-filled nunc tube.6. Refrigerate samples unl they can be shipped.7. If plasc pipe!es or syringes have been used,place them in a jar containing bleach. Allowto remain for ~ 10 minutes then dispose intored biohazard container.8. Send samples by courier to CMDN. CMDNwill forward culture samples to NATA andDNA-extracted samples to Dr. Pandey’slab. Samples should reach NATA as soonas possible as prolonged storage me canadversely affect results.Collecon of oropharyngeal swab for culture1. Wear N-95 mask and exam gloves.2. Place a culture!e on the end of the aluminumrod designed for this purpose3. Swab the oro-pharyngeal region.4. Place the swab in the tube and compress theend to release the transport media.5. Label with the elephant’s name, project ID #,collecon date, and facility abbreviaon.6. Refrigerate; ship to CMDN as soon as possible.CMDN will forward to NATA.Collecon of oropharyngeal swab for PCR andLAMP1. Wear N-95 mask and exam gloves.2. Place a sterile co!on swab on the end of aplasc extension rod.3. Swab the oro-pharyngeal region.4. Place the swab in the pre-labeled, buffer-fillednunc tube.5. Label with the elephant’s name, project ID #,facility abbreviaon, and collecon date.6. Refrigerate; ship to CMDN as soon as possible.9
Nepal Elephant Tuberculosis ulosisControl andManagement agemenAction Plan (2011-2015)2015)1.3 Descripon of DiagnoscMethodsa. The Elephant TB Stat-Pak ® test is a bloodanbody test. A reacve test indicatesprobable TB infecon. Confirmaon withother tests is ideal - when possible and asdiscussed above. A non-reacve test indicatesno infecon and a TB-free status at the meof tesng. The Elephant TB Stat-Pak ® test isused as the standard roune TB screeningtest for elephants in Nepal. The test isadministered by the TB Plan veterinarian.b. The DPP VetTB test is a blood anbody testused to confirm the Elephant TB Stat-Pak ® test.The Elephant TB Stat-Pak ® and DPP ® VetTBtests have been shown to be early predictorsof infecon and may be reacve monthsto years in advance of detecon by culture(Greenwald et al. 2009). This provides theopportunity to treat elephants before theyshed and infect other elephants, humans, orwildlife.c. Culture is used to detect elephants that areacvely shedding TB organisms. Respiratorysamples obtained by collecng trunkdischarges are submi!ed to a laboratorythat is qualified to test for mycobacteria(this requires specific procedures). A posiveculture confirms that an elephant is acvelyinfected. Posive isolates should be tested toconfirm if the organisms were M. tuberculosisor M. bovis. Drug sensivity tesng (DST) foran-tuberculosis drugs (isoniazid, rifampin,and ethambutol) should be performed.Note that culture has inherent limitaons.Failure to isolate the organism does notrule out infecon. False-negave resultsmay occur 1) if the elephant is infected butorganisms are not shed on the day of samplecollecon or 2) contaminated trunk washsamples cause other organisms to overgrow.False-negave results allow TB disease toprogress and the infected elephant is a sourceto spread TB to other elephants, humans,or wildlife. Culture is expensive and manysamples may be needed to detect infecon. InSweden, of 189 trunk wash cultures collectedfrom 5 elephants confirmed TB-infected onpostmortem, only 7 samples were posive(Moller et al. 2005).Newer technologies that prove to be morereliable, sensive, or cost effecve fordetecng TB organisms in trunk wash or othersecreons will be incorporated into this planas they become available.d. PCR (polymerase chain reacon) is a type ofnucleic acid amplificaon test (NAAT). NAATsfacilitate the idenficaon of pathogens byamplifying and detecng specific nucleicacid sequences. PCR assays to detect TB inelephants in Nepal are under invesgaon bythe Center for Molecular Dynamics-Nepal (seeKay et al. 2011).e. LAMP (loop-mediated isothermalamplificaon) assay is a type of NAAT thatallows detecon of trace amounts of DNAunder isothermal condions. It has been usedto diagnose TB in humans in Nepal (Pandey etal. 2008) and is under invesgaon for use inelephants.1.4 Segregaon ProceduresSegregaon is an important management tool.Segregaon from wild elephants and otherwildlife (e.g. rhino) is considered a priority toprevent TB transmission to endangered wildlifepopulaons. Segregaon can also minimizetransmission to other capve elephants and tohumans. The goal of segregaon is to preventhealthy animals from coming into contact withrespiratory droplets or other secreons frominfected elephants.10
Page 1: Nepal Elephant TuberculosisControl
Page 4: Copyright © and Published By:Gover
Page 11 and 12: SummaryThe Nepal Elephant Tuberculo
Page 13: AbbreviationsAREASBCCBZBZMCCNPCVLDL
Page 16 and 17: Mul angen print immunoassay (MAPIA)
Page 18 and 19: Part ITB Management
Page 20 and 21: Nepal Elephant Tuberculosis Control
Page 34 and 35: Part IIAction Plan
Page 48 and 49: Appendices

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