FIELD EVALUATION OF THE BIOEFFICACY OF CARBATIX 85 S WP AGAINST CATTLE TICKS UNDER NATURAL INFESTATIONS IN THE TRANSMARA WEST SUB-COUNTY OF KENYA

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FIELD EVALUATION OF THE BIOEFFICACY OF CARBATIX 85 S WP AGAINST CATTLE TICKS UNDER NATURAL INFESTATIONS IN THE TRANSMARA WEST SUB-COUNTY OF KENYA Martin, D. Muinde., Maisiba, C. Nyatich., Matheka, F. John., Mavuti S. Kitonga., Kivuvo, N. Muthini, Songok, K. Nathan., Kairu-Wanyoike Salome and Juma K. Ngeiywa Presented at The 50 th Kenya Veterinary Association s Annual Scientific Conference, Annual General Meeting and the 16 th World Veterinary Day Celebrations at Three Steers (Nairobi Pacific) Hotel, Meru: between 27 nd - 30 th April 2016 By Martin, D. Muinde, MAL&F State Department of Livestock, Vet. Labs - KABETE

1.1 Introduction Carbatix 85 S WP is based on carbaryl at 850 g/kg Carbaryl is the common name for the chemical 1-naphthyl N- methylcarbamate (EPA 1993) It belongs to a group of pesticides called carbamates (Tomlin 2000) It s a broadspectrum pesticide that has been used to control various ectoparasites of animals and humans (Schulze et al. 1992, Hoelscher et al. 1994, Burridge et al. 2002, Downs et al. 2002 ) It s been used widely to control R. (Bo.) microplus in many countries including Zambia, India, Jamaica, New Calendonia and Indonesia (Rawlins and Mansigh 1978, Sharma and Gupta 1982, Brun et al 1984, Basu and Halder, 1994 ) as well as the blue tick R. (Bo.) decoloratus (Koch) infesting cattle (Mathewson and Wilson 1976) Also used for control of small ruminant s, pet and poultry pests - fleas and lice

1.2 Carbatix 85 S WP Trial Objectives to specifically assess the therapeutic efficacy and ability of Carbatix 85 S WP to control high infestations of Rhipicephalus appendiculatus (Neumann, 1901), the brown ear tick as well as other tick species under the prevailing natural local conditions of Kenya using the manufacturer s proposed field usage rate of 2030 ppm or 0.2% at the Kenyan treatment regimen weekly use trials performed to satisfy the requirements of the Cattle Cleansing Act (CAP 358) and the PCPB Act (CAP 346 by the Central Vet Laboratories - Kabete under monitoring by the Director of Veterinary Services Vector Control Products Technical Group and the Pest Control Product s Board (PCPB) to have Carbatix 85 S WP gazetted and possibly registered for tick control in Kenya if it performs (during a test of equivalence) as well as or better than Steladone 300 EC, the only available registered Acetylcholinestarase Inhibitor Ixodicide in the Kenyan market

2.1 Materials and Methods. The local agent of the Registrant identified a suitable trial site at Mr. Samson Kisasi s Olosentu farm in Olosentu Sub-location, Olomismis Location, Kilgoris Division of the Transmara West Sub-County in the Narok County of the Republic of Kenya High tick loads obtainable :150-250 adult ticks per animal pre-treatment with ¾ being Rhipicephalus appendiculatus and theileriosis endemic Other tick species present include A.variegatum/gemma, Rhipicephalus (Boophilus ) decoloratus, Rhipicephalus evertsi and Hyalomma spp. Infrastructure cattle crush and adequate cattle enclosures (Bomas) Also provided a legal agreement indemnifying the DVS, PCPB and providing for disease diagnosis and treatment of clinical cases besides compensation for cattle in case of disease or accident

2.2. Trial lay-out and procedures. Pre-Trial cattle Selection. 50 pre-trial cattle selected on 09/12/2014, identified by ear tagging, vaccinated against FMD (Fotivax), BQ, Anthrax and de-wormed. Isolated to own boma and herded separately but not sprayed with a tickicide for 3 weeks under the care of a herdsman, watchman at night and a supervisor A Veterinarian and a Livestock Health Assistant/Technologist were engaged for clinical care. Mineral supplements were provided ad libitum Tick load verification done on 24/12/2014. Majority (26 0ut of 30 cattle which were examined) had the required 150-250 adult ticks

2.3 Trial Initiation Whole body tick counts were performed in situ on day -1 pre-treatment on the aforementioned 50 cattle Counts used to rank the animals- from one with highest to one with lowest tick numbers 21 experimental animals having moderate tick numbers selected and systematically allocated to 3 treatment groups of 7 animals each i.e negative control, Test and positive control groups. Carbatix 85 S WP was tested at the recommended Field Usage Rate of 2030 ppm as a Hand spray wash in comparison with Steladone 300 EC at the gazette FUR of 500ppm as the Reference tickicide. Its registered for tick control in Kenya Carbatix 85 S WP is carbaryl based. Sevin 85 S WP is the only similar product registered for tick control in Kenya but not available since the early 2000. Steladone 300 EC, a chlorfenvinphos-based E.C acaricide is the only related registered acaricide. Chosen due to having same mode of action as the test acaricide i.e both are acetylcholinesterase inhibitors. Both used briefly before moving to amitraz-based acaricides

2.4. Application of the acaricide. Each Test treatment animal was sprayed with 9 liters of freshly made Carbatix 85 S WP while those in the Reference Treatment group were sprayed with 10 liters of freshly made Steladone acaricidal wash using a Hobra Rocker Pump fitted with twin hoses and adjustable double spray cone nozzles. Spraying was done once every 7 days for a duration of 4 weeks Thorough cleaning of pump and crush in between treatments Animals in the negative control treatment group were left unsprayed through out the four-week experimental period.

2.5 The Hobra Rocker Pump

3.1 RESULTS AND DISCUSSION Whole body tick counts were done on all the 3 groups of animals on days 1, 3, 5 and 7 post-spraying (repeated weekly) Speciation, sexing and state of feeding of the ticks determined in situ on all the 3 treatment groups The negative control group was found to have the most ticks feeding to repletion cf to the others Engorged nymphs were observed in both treatment groups on days 5 and 7 posttreatment

Percentage Species contribution Fig 1. Tick species contribution over the carbatix 85 S WP experimental Period R. appendiculatus was most abundant:ranged 89.9-97.2% R. (Bo) decoloratus followe with btn 2.1-8.9% Amblyomma spp was last: ranged btn 0.7-1.9% 100 98 96 94 92 90 R. (Bo) decoloratus Amblyomma spp R. appendiculatus Ticks on negative control group representative of real picture of population 88 1 3 5 7 8 10 12 14 15 17 19 21 22 24 26 28 Days Post-Treatment

3.2 Efficacy determination The tick numbers obtained were used to calculate (efficacy) on these days using the formula of Drummond, Whetstone and Miller(1981) below: No. of ticks on control group No. of ticks on Treatment group X 100 No. of ticks on Control group

Percentage ttick Control Percentage Tick Control 3.3 EFFICACY (%) AGAINST THE ABUNDANT TICK SPECIES POST TREATMENT WITH CARBATIX 85 S WP Rhipicephalus appendiculatus Rhipicephalus (Boophilus) decoloratus 104 100 96 92 88 84 80 76 72 68 64 60 56 52 48 44 40 FIG 2: POST-TREATMENT R. APPENDICULATUS TICK CONTROL 0 1 3 5 7 8 10 12 14 15 17 19 21 22 24 26 28 Days Post-Treatment (Arrows indicate Days of Acaricide Spraying) CarbatIx 85 S WP Steladone 30% EC 104 100 96 92 88 84 80 76 72 68 64 60 56 52 48 44 40 FIG 3.: POST- TREATMENT R. (BO) DECOLORATUS TICK CONTROL 0 1 3 5 7 8 10 12 14 15 17 19 21 22 24 26 28 Days Post-Treatment (Arrows indicate Days of Acaricide Spraying) Carbatix 85 S WP Steladone 300 EC

Percentage Tick Control 3.4 OVERALL TICK CONTROL BY EACH OF THE ACARICIDES 100 96 92 FIG 4.: OVERALL PERCENTAGE TICK CONTROL POST- TREATMENT The Carbatix 85 S WP treatment group carried a slightly higher number of ticks on day 1 posttreatment than the Steladone 300 E.C animal treatment group but the difference was not statistically (P 0.05) significant. 88 84 80 76 72 68 64 60 56 52 48 44 40 0 1 3 5 7 8 10 12 14 15 17 19 21 22 24 26 28 Days Post Treatment (Arrows indicate Days of Acaricide Spraying) CarbatIx 85 S WP Steladone 300 EC It attained an overall therapeutic efficacy of 99.1% compared to 98.5% for Carbatix 85 S WP for day 1 i.e had a better tick knockdown effect Carbatix 85 S WP however, attained better overall efficacies of 97.9%, 90.2% and 77.4% compared to 91.4%, 76.6% and 58.1% for Steladone 300 EC on days 3, 5 and 7 posttreatment-attributed to its better performance against R. appendiculatus, the most abundant in the tick population

3.5 TOTAL NUMBERS OF ENGORGED NYMPHS PER TREATMENT GROUP POST- ACARICIDE SPRAYING WEEK 1 WEEK 2 WEEK 3 WEEK 4 TREATMENT DAY 5 DAY 7 Carbatix 85 S WP 0 37 Steladone 300 EC 25 249 Carbatix 85 S WP 3 54 Steladone 300 EC 32 348 Carbatix 85 S WP 1 40 Steladone 300 EC 42 359 Carbatix 85 S WP 11 156 Steladone 300 EC 32 334 Engorged R. appendiculatus nymphs were observed and enumerated on days 5 and 7 Nymphs feeding on ECF sick animals drop and moult Resulting adults infective and will transmit the disease The Steladone 300 E.C animal treatment group always harbored more engorging/engorged nymphs than the Carbatix 85 S WP treatment group

4.0 Conclusion and Recommendation. Neither the test nor the reference acaricide attained the originally required minimum preregistration tick control efficacy benchmarks of between 99%, 96% and 94% for organophosphate or chlorfenvinphos-based Ixodicides in Kenya on days 3, 5 and 7 post-spraying due to emerging resistance by the Rhipicephalus (Boophilus) decoloratus population in the area (Lab results) Carbatix 85 S WP was however found to be equivalent in tick knock-down to Steladone 300 E.C but clearly superior in performance (therapeutic efficacy and repellent action) against both adult and immature Rhipicephalus appendiculatus (Neumann, 1901) ticks than Steladone 300 E.C while the latter was more efficacious against Rhipicephalus (Boophilus) decoloratus (Koch, 1844) the blue tick Carbatix 85 S WP will therefore offer more satisfactory control of high infestations of both adult and immature Rhipicephalus appendiculatus and other tick species infesting cattle when applied once weekly compared to Steladone 300 E.C On this strength then, the acaricide was gazetted for tick control in the country as a hand spraywash at the tested usage rate.

5.0 Acknowledgements We wish to acknowledge the past work of W.J. Roulston, R.H. Wharton and J. Nolan of CSIRO and Dr. Rinkanya, F. G formerly of the Central Veterinary Laboratories-Acaricide Laboratory, Kabete who were responsible for much of the work behind development of the study protocol used in this work. We also thank the following:- M/S Zhejiang E-Tong Chemical Company Limited, Shanghai, China and Highchem Marketing Limited, Essentials Division Nairobi, Kenya for sponsoring this trial Dr. Jonathan Orengo of Highchem Marketing Limited, Nairobi, Kenya for ascertaining that all the required supplies were provided on time Mr. Samson Kisasi for offering the trial site and his kith and kin especially Mr. Samson Kinne for allowing use of their animals for the trial Mr. Ole Ng inai Kukuu and Ole Naigeyo Kukuu, among others, for the offer of experimental animals pastures Mr. Lucas M. Agan for providing health care to experimental animals Mr. David Kisasi (watchman), Joseph L. Tieparuso, Mr. Gideon Kinne, Joseph Naigeyo, Edward Naigeyo, David Karandi (Trial animals Herdsmen) and Mr. Peter Sumpel for maintaining site cleanliness. The Director of Veterinary Services for allowing this presentation All of you for listening to me

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