USAHA Parasitic and Vector Borne Diseases … · USAHA Parasitic and Vector Borne Diseases...

Dr. John Goolsby et al.USAHA Parasitic and Vector Borne Diseases Committee Meeting

San Diego, CA, 18 October 2018

1

Research Update

John Goolsby, Ph.D.Collaborators: Nirbhay Singh, Beto Perez de Leon,

Poncho Ortega, Tyler Campbell, Dave Hewitt, Randy DeYoung, Lisa

Zoromski,Clint Hoffman, David Shapiro,

Alex Racelis

U.S. Dept. of Agriculture, Agricultural Research Service, Cattle Fever Tick Research Laboratory, Edinburg, TX;

Guru Angad Dev Veterinary and Animal Sciences University, Punjab, India

USDA-ARS Knipling Bushland U.S Livestock Insects Research Laboratory, Kerrville, TX;

East Foundation, San Antonio, TX

Texas A&M Kingsville- Cesar Kleberg Wildlife Institute, Kingsville, TX

USDA-ARS, College Station, TX & Byron, GA

University of Texas, Rio Grande Valley, Edinburg, TX

Update

• Nilgai home range

• Nilgai lures

• Lab and Barn test of tick nematodes

• Nilgai sprayer

• Nilgai latrine ecology

• Exploration CFT native range for BC agents

Cattle Fever Tick (CFT)

Rhipicephalus (=Boophilus) microplus

• One host tick

• Hosts: Bovidae & Cervidae

• Transmit bovine babesiosis

• Resistant to most acaricides

• Worldwide distribution in

tropical, subtropical and

warm temperate climates

• Major limitation to cattle &

milk production

Cattle Fever Tick Hosts Texas & Mexico

Nilgai

Deer

Cattle • Acaricides, vaccines

• Medicated corn feeders

• No known methods for

treatment

What has changed?

Tick is resistant to most acaricides

Exotic weeds favor survival of tick

A new, highly mobile, host animal of the tick

is now widespread

Nilgai Biology• Members of the Bovidae

• Closely related to cattle

• Native to India

• Cows – 600 lbs. Bulls – 800 lbs.

Nilgai Home Range Study

• Collaborators: Hewitt, Foley, TAMUK; Campbell and

Ortega – East Foundation

• Funding: Texas Animal Health Commission

• Location: East Foundation – El Sauz, Willacy Co.

• 30 adult nilgai collared with GPS tracking devices (Lotek)

50/50 bulls/cows

• Data: home range, max distance, effect of helicopters,

physical barriers (roads & farmland)

Foley, A.M. Goolsby, J. A., Alfonso Ortega-S., Jr. J. Alfonso Ortega-S., Adalberto Perez de

Leon, David G. Hewitt, Tyler A. Campbell, and Nirbhay K. Singh . Movement Patterns of

Nilgai Antelope (Boselaphus tragocamelus) in South Texas: Implications for Cattle Fever

Tick Management. Journal of Wildlife Ecology. Preventive Veterinary Medicine (in review).

HWY 186

Port Mansfield

King Ranch

El Sauz

farmland

Females have > home range

Young females have largest

home range by far!

Nilgai Lure Study

• Collaborators: Hewitt, TAMUK; Campbell

and Ortega – East Foundation; Singh, Guru

Angad Dev Veterinary University

• Nilgai attractants would be useful for

developing CFT treatments

• Nilgai form communal latrines

• Nilgai prefer to make latrines on offal

• Testing synthetic offal (Screw worm lure,

and bovine volatile fatty acids (VFA)

Nilgai Latrine Offal – Induces Latrine

Screwworm Lure – Offal Mimic

Nilgai Bull at Lure

Nilgai Lure Study Results

• Offal the most attractive

• Screwworm lure the only other lure that

induced latrine formation

• High rates of Screwworm lure being tested

at East Foundation, Santa Rosa

Goolsby, J.A., Singh, N. K., Ortega-S. A., Jr., Hewitt, D.G., Campbell, T.A,

Wester, D. and Pérez de León, A.A.. Comparison of natural and artificial odor

lures for nilgai (Boselaphus tragocamelus) and white-tailed deer (Odocoileus

virginianus) in South Texas: developing treatment for cattle fever tick eradication.

International Journal for Parasitology: Parasites and Wildlife. (In Press).

Update


• Nilgai lures


• Nilgai sprayer

• Thermal UAV density estimates & harvest



Attract and Treat at Latrines

• Collaborators: Clint Hoffman, Ag Engineer,

USDA-ARS College Station; David Shapiro,

Nematologist, USDA-ARS, Byron, GA; Hewitt,

TAMUK; Campbell and Ortega – East

Foundation; Singh, Guru Angad Dev Veterinary

University

• Sensor activated ultra quiet sprayer

• Use nematode, native Steinernema riobravae to

control CFT, acceptable for USFWS.

• Funding USDA-APHIS-VS Microgrant Program

Nilgai Sprayer

Lure

Nilgai at Latrine

Microjet

Nilgai sprayer at fence crossing

Mist from sprayer

Nilgai sprayer at latrine

Nematode spray

tank & pump

Update


• Nilgai lures


• Nilgai sprayer




Lab and barn test of tick nematodes

• Heterorhabita floridensis &

Steinernema riobravae highly

effective in lab assays

• Whole animal treatment study at

1X completed with moderate

efficacy. Plan to repeat at 10X

• Steinernema riobravae native to South

Texas and Northeastern Mexico and has

high heat tolerance

• S. riobravae is commercially available

from BASF Company

Nematode Infection of CFT

1 day after nematode application• Normal cattle fever tick

7 days after nematode

application

control

Nematode Nematode

concentration

(IJs/dish)

Av. tick wt. ±

SE (mg)

Av. egg mass

wt. ± SE (mg)

RI ± SE %IO ± SE

Steinernema

riobrave

1250 293.9 ± 12.5 100.8 ± 11.5 0.34 ± 0.03b 37.27 ± 6.94

2500 354.8 ± 11.3 12.3 ± 6.5 0.04 ± 0.02b 93.03 ± 3.93

5000 352.3 ± 6.8 0 ± 0 0 ± 0b 100 ± 0

S. carpocapsae 1250 334.1 ± 14.7 129.2 ± 11.4 0.37 ± 0.03b 31.11 ± 5.27

2500 348.9 ± 12.8 26.1 ± 10.0 0.07 ± 0.02b 86.49 ± 5.23

5000 379.5 ± 11.7 16.2 ± 12.3 0.04 ± 0.02b 93.47 ± 4.76

S. feltiae 1250 333.8 ± 17.5 142.2 ± 12.0 0.43 ± 0.03a 21.56 ± 6.11

2500 346.7 ± 10.1 165.2 ± 11.8 0.48 ± 0.03 12.61 ± 5.89

5000 352.7 ± 8.9 160.5 ± 14.6 0.45 ± 0.04 17.47 ± 7.30

Heterorhabditis

bacteriophora

1250 342.6 ± 17.5 82.3 ± 13.9 0.23 ± 0.04b 57.19 ± 7.57

2500 333.2 ± 13.8 15.7 ± 6.3 0.06 ± 0.02b 89.82 ± 4.46

5000 359.2 ± 8.0 2.9 ± 2.0 0.008 ± 0.005b 98.53± 1.02

H. indica 1250 318.3 ± 14.4 117.5 ± 17.5 0.36 ± 0.05b 34.47 ± 9.36

2500 349.9 ± 13.8 21.4 ± 10.1 0.06 ± 0.03b 88.33 ± 5.94

5000 378.0 ± 10.4 1.0 ± 1.0 0.002 ± 0.002b 99.57 ± 0.42

H. floridensis 1250 321.7 ± 16.7 1.45 ± 1.45 0.003 ± 0.003b 99.40 ± 0.59

2500 350.5 ± 9.2 0 ± 0 0 ± 0b 100 ± 0

5000 354.1 ± 12.0 0 ± 0 0 ± 0b 100 ± 0

Control - 312.3 ± 13.7 172.2 ± 12.8 0.55 ± 0.03 0.0

Effect of various nematode infective juveniles (IJs) on reproductive parameters of Rhipicephalus

(Boophilus) microplus engorged females following 72 h exposure on filter paper

RI- Reproductive index; % IO- Percentage inhibition of oviposition; a(P < 0.05); b(P < 0.01)

3.00 3.25 3.50 3.750

5

10 SR

SC

SF

HB

HI

HF

Log Conc

Pro

bit M

ort

ality

Dose mortality curve of R. microplus against nematode species on 72 h exposure

Effect of Steinernema & Heterorhabditis

IJs against Rhipicephaus microplus

• In filter paper test exposure tests, 72 hours H. floridensis caused

100% at 2500 IJ/ml, followed by S. riobravae 100% at 5000 IJ/ml

• In immersion tests, H. floridensis caused 100% mortality at 5000

IJ/ml with an immersion for above 30 sec period. However, other

EPN did not cause high mortality in any immersion time (max. of

40% for H. indica for 30 min immersion).

• A significant effect on the reproductive parameters of R.

microplus female ticks was recorded for all EPNs.

• Hatchability percent of 60-70 was recorded in S. riobrave exposed

groups whereas, no effect was seen in eggs of other groups.

Update


• Nilgai lures


• Nilgai sprayer




Thermal UAV

• Can detect nilgai under

dense brush canopy

• 1km meandering

transects surveys in

Cameron Co. = 1: 88

• How does density

compare to 1: 44 ratio

at East Foundation?

Goolsby, J. A., J. Jung , J. Landivar, W. McCutcheon, R. Lacewell, R. Duhaime, D. Baca, R. Puhger, H. Hasel, K. Varner, B. Miller,

A. Schwartz & A. Perez de Leon. 2016. Evaluation of Unmanned Aerial Vehicles (UAVs) for detection of cattle in the Cattle Fever

Tick Permanent Quarantine Zone Subtropical Agriculture and Environments 67:24-27.2016

Update


• Nilgai lures


• Nilgai sprayer




Latrine Ecology

• Study with CKWRI, M.S. student

• Funded by Las Huellas wildlife organization

• How many latrines per acre

• How many nilgai visit each latrine (molecular

analysis of dung)

• Tie-in to nilgai sprayer

Update


• Nilgai lures


• Nilgai sprayer




Biological control of cattle fever ticks

.

J. Goolsby1, F. Guerrero2, J. Kashefi3, L, Smith3, A. Racelis4, D. Amalin5, M.J. Flores5, N. Abes5; K. Verly5; N. Singh6; P. Azhahianambi7, A. Jesudasan8, P. DeBarro9, A. Sheppard9, K. Wyckuys10, J. Liu11, A. Schwartz12, H. Hasel13, K. Varner13, & A. Pérez de León2

1United States Dept. of Agriculture, Agricultural Research Service, Knipling Bushland U.S. Livestock Insect Pest Research Laboratory, Cattle Fever Tick Unit, Edinburg, TX, [email protected]; 2USDA-ARS Knipling Bushland U.S. Livestock Insect Pest Research Laboratory, Veterinary Pest Genomics Center, Kerrville, TX; 4USDA-ARS European Biological Control

Laboratory, Montpelier, France; 4Univ. of Texas, Rio Grande Valley; 5De La Salle University, Manila, Philippines; 13Dept. of Veterinary Parasitology, Guru Angad, Dev Veterinary and Animal Sciences Univ., Punjab, India, 7Dept. of Veterinary Parasitology, Madras Veterinary College, TANUVAS, Chennai, India; 8Madras Christian College, Chennai, India; 9CSIRO Health &

Biosecurity, Brisbane & Canberra, Australia; 10International Center for Tropical Agriculture, Hanoi, Vietnam; 11Hebei Normal University, Shijiazhuang, Hebei, P.R. China; 12Texas Animal Health Commission, Austin, TX; 13USDA-APHIS, Cattle Fever Tick Eradication Program, Laredo, TX.

The southern cattle fever tick, Rhipicephalus microplus has several important hosts including

domestic cattle, whitetail deer and Indian nilgai antelope. Biological control could potentially

reduce populations of tick in Mexico and reduce the risk of invasion into the U.S. along the Rio

Grande. Biological control could be integrated with current cattle fever tick eradication

programs.

Cattle Fever Ticks Exploration in Native Range

Wildlife Hosts of

Tick

White-tailed Deer Exotic Indian Nilgai

Fig. 1. Native and introduced ranges of cattle fever ticks (Rhipicephalus spp.) Red =

introduced range of R. microplus, Pink = R. microplus eradicated, Blue = native range

of R. annulatus, Purple = introduced range, Green = native range of R. microplus,

Yellow = native range of R. australensis. Stars = search locations for biocontrol

agents.

1

2

Fig. 2. (A) Vial of 500 questing larvae to be glued to the heifers for release, (B) Tag cement used to glue the vials of questing tick larvae to the heifers, (C) The heifers being run into the working chute for the questing tick larvae to be released on them, (D) Glue is placed on the vial of questing larvae prior to being placed on the top of the heifer’s shoulder, (E) Questing tick larvae vial being placed on the top of the shoulder, (F) The heifers in the pen after they were manually infested with Rhipicephalus annulatus and R. microplus questing larvae

3

D

A B

C

E F

Fig. 4. (A) The two heifers in the reduced pen for the duration of the tick drop to minimize the area needed to search for ticks. (B) Nearly engorged Rhipicephalus microplus on the side of one of the heifers; (C)

Searching the pen for fallen engorged female ticks; (D) Fallen engorged female ticks found in the chute; (E) The field collection vial used to store the ticks in until we are back in the lab; (F) engorged ticks are held in vials in humidified aquarium for emergence of parasitoids

E F

C D

A B

Tick Exposures to Collect Parasitoids

Potential Biocontrol Agents

These pictures shows the predator mites Anystis jabanica and Walzia australica

which are known to predate on R. microplus in Indonesia/Australia. On right is a

steinernemid nematode which can be used as a tick biocides. Other CFT

specialists may occur in the native range.

Fig. 6. (A) Release of questing larvae onto test plant. (B) Questing larvae on topside of leaf. (C) Time lapse photography camera for observing predation of questing larvae on plant. (D) Battery and LED red light used for night time observation. (E) Day time photo of larvae on underside of leaf. (F) Night time observation of larvae

with spider in background.

A B

C D

E F

Predation Questing Larvae

Fig 7. Collection of soil for nematode assays from (A) tika deer bedding areas (India). (B) Water buffalo emerging from wallow (Philippines). (C) Fully engorged cattle fever ticks placed on moist soil in Ziploc to assay for nematodes. (D) Entomopathogenic nematodes in fly ovaries of a fly.

A B

C D

Soil-Dwelling Predators

These pictures show the tick parasitoid Ixodiphagous hookeri, a parasitoid of

the tick, Ixodes scapularis. We hypothesize that a similar specialist parasitoid

occurs in the native range of cattle fever tick. On right is Ixodiphagous

aethes, a newly described species from India. Its tick host is unknown.

White-tailed deer and other exotic wildlife can be hosts of CFT and treatment of tick infestation

is difficult. Nilgai (native to India) are highly mobile animals and have the potential to spread

ticks beyond the border with Mexico and within temporary quarantine zones in Texas.

Currently, no treatment is available for CFT on nilgai.

Goolsby, J.A., Mays, D. T., Schuster G. L., Kashefi J., Smith L., Amalin D., M. Cruz-Flores, Racelis A., and Pérez de León, A.A. 2016.

Rationale for Classical Biological Control of Cattle Fever Ticks and Proposed Methods for Field Collection of Natural Enemies.

Subtropical Agriculture and Environments. 66:7-15.

Goolsby, J. A., Guerrero F. D., Gaskin J., Bendele K. G., Azhahianambi P., Amalin D., Flores-Cruz M., Kashefi J., Smith L., Racelis A.,

Saini R. K., and Perez de Leon A. 2016. Molecular comparison of cattle fever ticks from native and introduced ranges with insights into

optimal search areas for classical biological control agents. Southwestern Entomologist 41: 595-604.

Distributions of cattle fever ticks

.









programs.


Wildlife Hosts of

Tick








Predation Questing LarvaeSoil-Dwelling Predators









Fig. 1. Native and introduced ranges of cattle fever ticks (Rhipicephalus spp.) Red = introduced range of R.

microplus, Pink = R. microplus eradicated, Blue = native range of R. annulatus, Purple = introduced range,

Green = native range of R. microplus, Yellow = native range of R. australensis. Stars = search locations for

biocontrol agents.

Wasp inserts egg tick nymphEgg hatches and wasp larvae begins feeding

Wasp larvae completes development

Tick mummifies and next generation of wasps emerge

Multiple wasps emerge from dead tick

Adult wasps search for ticks

Life cycle of the tick parasitoid Ixodiphagus hookeri on Ixodes ricinustick

Biological control of cattle fever ticks

.









programs.


Wildlife Hosts of

Tick


Fig. 1. Native and introduced ranges of cattle fever ticks (Rhipicephalus spp.) Red =

introduced range of R. microplus, Pink = R. microplus eradicated, Blue = native range

of R. annulatus, Purple = introduced range, Green = native range of R. microplus,

Yellow = native range of R. australensis. Stars = search locations for biocontrol

agents.

1

2

Fig. 2. (A) Vial of 500 questing larvae to be glued to the heifers for release, (B) Tag cement used to glue the vials of questing tick larvae to the heifers, (C) The heifers being run into the working chute for the questing tick larvae to be released on them, (D) Glue is placed on the vial of questing larvae prior to being placed on the top of the heifer’s shoulder, (E) Questing tick larvae vial being placed on the top of the shoulder, (F) The heifers in the pen after they were manually infested with Rhipicephalus annulatus and R. microplus questing larvae

3

D

A B

C

E F

Fig. 4. (A) The two heifers in the reduced pen for the duration of the tick drop to minimize the area needed to search for ticks. (B) Nearly engorged Rhipicephalus microplus on the side of one of the heifers; (C)

Searching the pen for fallen engorged female ticks; (D) Fallen engorged female ticks found in the chute; (E) The field collection vial used to store the ticks in until we are back in the lab; (F) engorged ticks are held in vials in humidified aquarium for emergence of parasitoids

E F

C D

A B







Fig. 6. (A) Release of questing larvae onto test plant. (B) Questing larvae on topside of leaf. (C) Time lapse photography camera for observing predation of questing larvae on plant. (D) Battery and LED red light used for night time observation. (E) Day time photo of larvae on underside of leaf. (F) Night time observation of larvae

with spider in background.

A B

C D

E F

Predation Questing Larvae

Fig 7. Collection of soil for nematode assays from (A) tika deer bedding areas (India). (B) Water buffalo emerging from wallow (Philippines). (C) Fully engorged cattle fever ticks placed on moist soil in Ziploc to assay for nematodes. (D) Entomopathogenic nematodes in fly ovaries of a fly.

A B

C D

Soil-Dwelling Predators









Goolsby, J.A., Mays, D. T., Schuster G. L., Kashefi J., Smith L., Amalin D., M. Cruz-Flores, Racelis A., and Pérez de León, A.A. 2016.

Rationale for Classical Biological Control of Cattle Fever Ticks and Proposed Methods for Field Collection of Natural Enemies.

Subtropical Agriculture and Environments. 66:7-15.

Goolsby, J. A., Guerrero F. D., Gaskin J., Bendele K. G., Azhahianambi P., Amalin D., Flores-Cruz M., Kashefi J., Smith L., Racelis A.,

Saini R. K., and Perez de Leon A. 2016. Molecular comparison of cattle fever ticks from native and introduced ranges with insights into

optimal search areas for classical biological control agents. Southwestern Entomologist 41: 595-604.

Summary

• Develop method to treat CFT on nilgai to

aid in eradication of CFT from Cameron

and Willacy Counties.

• Develop biocontrol methods to reduce CFT

populations in Mexico along border

USAHA Parasitic and Vector Borne Diseases … · USAHA Parasitic and Vector Borne Diseases...

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Transcript of USAHA Parasitic and Vector Borne Diseases … · USAHA Parasitic and Vector Borne Diseases...