THERIOCENOLOGY

ELECTROEJACULATION OF THE COYOTE

M. L. Brussa, J. S. Green and J. N. Stellflug
U. S. Department of Agri cul ture, ARS
U. S. Sheep Experi ment Stati onb
Duboi s, ID., 83423
Recei ved f or Publ i cati on: January 24, 1983
Accepted: May 12, 1983
ABSTRACT
Two el ectroej acul ators were used to col l ect semen f rom 40 adul t
mal e coyotes. The most ef f ecti ve apparatus used a two- ri ng rectal probe
and an AC vol tage of 18 (Vrms) at 1000 Hz. Wi th thi s ej acul ator, 11 of
15 coyotes produced a sati sf actory semen sampl e, whi ch averaged 0. 9 ml
i n vol ume and 70 mi l l i on spermatozoa per ml .
INTRODUCTION
Studi es of reproducti ve physi ol ogy and eval uati on of anti f erti l i ty
drugs f or mal e coyotes woul d be enhanced by a sui tabl e means f or
col l ecti ng semen sampl es. Due to thei r wi l d temperament, mani pul ati ve
means are not usual l y rel i abl e i n produci ng semen sampl es f rom mal e
coyotes, nor i s usi ng a teaser bi tch and arti f i ci al vagi na f easi bl e f or
most i ndi vi dual s. El ectroej acul ati on of anestheti zed coyotes appears to
be the most practi cal method of obtai ni ng semen f rom thi s speci es.
An el ectroej acul ator f or use wi th anestheti zed dogs has been
descri bed (1) and has been used f or coyotes (2). In the study reported
here, we i nvesti gated the ef f i cacy of the dog el ectroej acul ator and a
newl y desi gned el ectroej acul ator.
NATERIALS AND METHODS
The 40 coyotes used i n thi s study were adul t mal es housed i n
kennel s wi th i ndi vi dual outdoor runs and i ndoor boxes. Water and f ood
were provi ded ad l i bi tum. Pri or to el ectroej acul ati on, the ani mal s were
gi ven 0. 5 mg atropi ne and ei ther 5 mg acetyl promazi ne (ol der- desi gn
ej acul ator) or 25 mg xyl azi ne (newer- desi gn ej acul ator) IM. Af ter the
tranqui l i zer had taken ef f ect, a soapy water enema was admi ni stered.
Ani mal s were then anestheti zed wi th 4% thi amyl al sodi um IV. An attempt
was made to empty the bl adder of uri ne ei ther by manual compressi on
(ol der- desi gn el ectroej acul ator) or catheteri zati on (newer- desi gn
el ectroej acul ator) to reduce uri ne contami nati on of semen.
' Present address Department of Physi ol ogi cal Sci ences
and repri nts: School of Veteri nary Medi ci ne
Uni versi ty of Cal i f orni a Davi s, CA 95616
bi n cooperati on wi th the Uni versi ty of I daho, Moscow, I daho. The
techni cal assi stance of J . Ri ch, R. Adai r, M. Luther, R. A. Woodruf f ,
and W. Bowers is gratef ul l y acknowl edged. Menti on of a propri etary
product does not consti tute a guarantee nor i mpl y approval or
di sapproval by USDA.
3ULY 1983 VOL. 20 NO. 1 53
THERIOGENOLOGY
Coyotes were used f or compari ng the two el ectroej acul ators i n
J anuary, whi ch was duri ng the normal breedi ng season (3). The ol der-
desi gn ej acul ator was tested i n 1980 and the newer- desi gn ej acul ator was
tested i n 1982. In addi ti on, prel i mi nary testi ng of the newer- desi n
ej acul ator was perf ormed i n
1981 duri ng l ate March and earl y Aprl , 3
whi ch was l ate i n the breedi ng season (3).
The ol der- desi gn el ectroej acul ator (1) was purchased f rom Ni chol son
Manuf acturi ng, Denver, CO. Thi s apparatus used one cutaneous el ectrode
appl i ed i n the l umbosacral area and a rectal probe consi sti ng of a
copper rod 15 cm l ong and 0. 6 cm i n di ameter. The posteri or 12. 5 cm of
the rectal probe was covered wi th pl asti c i nsul ati ng tape. The
ej acul ator produced a si ne wave vol tage of O- 50 Vrms (root mean square
vol ts) at 60 Hz and a maxi mum current of 0. 25 amps. Af ter pl acement of
the el ectrodes, 30 Vrms were appl i ed f or 4- 10 set wi th 5- 10 set rest
peri ods i n between. As many as 10 cycl es were used i n attempti ng to
obtai n an ej acul ate.
The newer el ectroej acul ator was desi gned to al l ow appl i cati on of a
l arge vari ety of vol tages, f requenci es, and wavef orms. The key component
was a f uncti on generator (Model 3010, B+K Preci si on Dynascan Corp. ,
Chi cago, IL) whi ch had an output of O- 20 V at 0. 1 Hz - 1 MHz wi th the
choi ce of a tri angl e, square, or si ne wave. The output of the f uncti on
generator was connected to the i nput of an audi o ampl i f i er (Power Pl us
120, Fanon- Couri er Corp. , Pasadena, CA) whi ch coul d have i ncreased the
vol tage to as hi gh as 70 Vrms i n the f requency range of 30 Hz - 20
KHz. The output of the ampl i f i er was connected to a si mpl e control l er
whi ch provi ded no, hal f - wave, or f i l tered f ul l - wave recti f i cati on (f or
testi ng AC, pul sed DC, or conti nuous DC currents, respecti vel y). The
control l er al so provi ded connecti ons f or an osci l l oscope and a rectal
probe, and an on- of f swi tch f or the probe. The osci l l oscope (Model RM-
45A, Tektroni x Corp. , Beaverton, OR) al l owed conti nuous moni tori ng of
the f requency, vol tage, and wavef orm of the probe current. The
assembl ed apparatus i s depi cted i n Fi gure 1.
Two rectal probes f or the newer ej acul ator were constructed f rom
cl ear acryl i c pl asti c. The probes were 35 cm l ong and 2. 5 cm i n
di ameter, the l atter di mensi on bei ng the average di ameter of coyote
scats (4). The anteri or ends of both probes were rounded f or easy
i nserti on. One probe, desi gnated as the two- ri ng probe, had a sol i d
pl asti c anteri or segment 3. 8 cm i n l ength wi th a ci rcumf erenti al copper
ri ng 1. 0 cm i n wi dth and 0. 16 cm thi ck attached to the posteri or end of
i ts l ateral surf ace. The anterior segment was gl ued to an acryl i c
pl asti c tube whi ch was 1. 6 cm i n di ameter and 41 cm i n l ength and served
as a handl e. An i nsul ated wi re was sol dered to the copper ri ng and
passed through the handl e. The posteri or segment consi sted of pl asti c
tubi ng, 2. 5 cm i n di ameter and 25 cm i n l ength. A copper ri ng,
i denti cal to that on the anteri or segment, was gl ued on the anteri or end
of the l ateral surf ace of the posteri or segment. An i nsul ated wi re was
sol dered to the copper ri ng and threaded through the posteri or
segment. The probe was assembl ed by passi ng the handl e through the
posteri or segment. The di stance between the ri ngs coul d be vari ed by
movi ng the handl e, whi ch had cal i brati on marks at 1 cm i nterval s, f rom
the rear of the posteri or segment. Later, a hol l ow pl asti c i nsert, 2. 5
cm i n di ameter by 4. 0 cm l ong, was pl aced between the anteri or and
posteri or surf aces to f i x the ri ng gap at 4. 0 cm (edge to edge). Thi s
probe and i ts components are shown i n Fi gure 2.
54
JULY 1983 VOL. 20 NO. 1
THERIOGENOLOGY
The other probe, desi gnated as the f our- stri p probe, consi sted of a
pi ece of sol i d pl asti c 2. 5 cm i n di ameter and 4. 4 cm l ong gl ued to a
hol l ow pl asti c tube 2. 5 cm i n di ameter and 30 cm l ong.
Four copper
stri ps 0. 16 cm thi ck, 0. 64 cm wi de, and 15 cm l ong were gl ued to the
pl asti c tube 90" apart. Pai rs of stri ps 180" apart were connected
el ectri cal l y,
and wi res to each of the two pai rs of St r i ps w er e
connected to the control l er.
The two- ri ng probe was usual l y i nserted so that the posteri or ri ng
was 3 cm anteri or to the anus and the anteri or ri ng was 4 cm anteri or to
the posteri or ri ng. The f our- stri p probe was i nserted so that the
posteri or edges of the stri ps were 3 cm anteri or to the anus. These
di stances were determi ned by external l y measuri ng the di stances f rom the
anus to the anteri or and posteri or edges of the sacrum. Si nce the
pel vi c, pudendal , and sacral nerves whi ch control ej acul ati on ori gi nate
i n the sacral regi on, maxi mal sti mul ati on was expected to oc c ur when
metal el ectrodes i n the rectum spanned thi s regi on (5).
Fi gure 1. El ectroej acul ator el ectri cal apparatus. The topnost uni t i s
the control l er whi ch connects the rectal probe to the apparatus and
al l ows modi f i cati on of the wavef orm.
The si gnal generator i s under the
control l er and provi des a l ow- power si gnal whi ch i s boosted by the audi o
ampl i f i er, whi ch i s under the si gnal generator.
The bottom uni t i s the
osci l l oscope whi ch i s used to moni tor the el ectri cal output to the
rectal probe.
JULY 1983 VOL. 20 NO. 1
55
THERIOGENOLOGY
Fi gure 2. Two- ri ng probe. The sol i d anteri or segment
to the hol l ow handl e ( B) whi ch was movabl e through the
( A) was attached
hol l ow posteri or
segment ( C) . The cal i brati on marks on the posteri or segment were used
to j udge the depth of i nserti on. When 4 cm was f ound to be an
acceptabl e i nter- ri ng di stance, a pl asti c i nsert ( D) was used between
the ri ngs.
Af ter i nserti on, the two- ri ng probe was swi tched on f or 2- 4 set and
of f f or 2- 4 set and conti nued f or UD to 10 cvcl es i f no ei acul ate was
obtai ned. A si ne wave of 18 Vrms at' 1000 Hz was tri ed i ni ti al l y on al l
ani mal s whi ch were used f or testi ng the newer apparatus. Thi s vol tage
was chosen as bei ng strong enough to overcome the resi stance of mucus
and f eces i n the rectumbut weak enough to al l ow a current f l owof l ess
than 0. 25 amp to provi de saf ety f or the coyote and the operators.
Li kewi se, the f requency of 1000 Hz provi ded saf ety dnd good current
penetrati on i nto ti ssues ( 6) . Other vol tages or wavef orms or the f our-
stri p probe were used i n the prel i mi nary tri al s i n 1981 onl y i f the
i ni ti al attempts produced no semen. For the tri al s i n 1982, onl y a si ne
wave at 18 Vrms and 1000 Hz was used.
Semen was col l ected i n pl asti c bags and exami ned di rectl y under the
mi croscope f or moti l i ty at 100 X magni f i cati on. Fewer than 50%moti l e
spermatozoa was scored a 1, 50- 90X moti l e was scored 2, and greater than
90% moti l e was scored a 3. Semen was di l uted wi th sal i ne, and
spermatozoa were counted i n a hemocytometer at 400 X magni f i cati on.
RESULTS
Obtai ni ng adequate semen sampl es wi th the ol der system of
cutaneous- rectal l eads was margi nal l y successf ul ( 2 out of 7, Tabl e
1) .
In addi ti on, the muscul ar ri gi di ty caused by thi s apparatus was
j udged to be greater than wi th the two- ri ng probe system.
Obtai ni ng useabl e semen sampl es wi th the two- ri ng probe duri ng
J anuary was si gni f i cantl y more successf ul ( p < 0. 05, Chi - square) than
wi th the cutaneous- rectal l eads ( Tabl e 1) . Moti l i ty scores were l ower
f or semen sampl es obtai ned wi th the cutaneous- rectal l ead systemthan
wi th the two- ri ng rectal probe system( p < 0. 001 by unpai red t- test) .
JULY 1983 VOL. 20 NO. 1
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THERIOGENOLOGY
When semen was obtai ned, i t was usual l y on the second through f i f th
appl i cati on of vol tage. If an ej acul ate was not obtai ned by the f i f th
appl i cati on of vol tage, f urther appl i cati ons were usual l y f rui tl ess.
Onl y parti al peni l e erecti on was obtai ned, probabl y because of the short
durati on of the vol tage.
Other than muscul ar ri gi di ty, the onl y si de
ef f ect was an occasi onal smal l amount of bl ood on the probe. Recovery
f rom the anestheti c was routi ne f ol l owi ng el ectroej acul ati on.
The resul ts of prel i mi nary tri al s i n 1981 wi th the newer ej acul ator
are al so presented i n Tabl e 1. There were 28 tri al s usi ng 18 coyotes,
and no i ndi vi dual coyote was used f or more than three tri al s. The data
f or 1981 i n Tabl e 1 represent the most f avorabl e resul t f or the tri al s
wi th each coyote. In those tri al s i n whi ch no semen was obtai ned wi th
the standard vol tage parameters , varyi ng the vol tage ampl i tude (up to 30
Vrms), f requency (DC, 25 or 100 Hz), or wave f orm (tri angl e or square)
di d not produce an ej acul ate. The f our- stri p probe was used onl y once
and caused a current f l ow of more than 0. 25 amp; theref ore, i t was not
used agai n.
DI SCUSSI ON
The two- ri ng probe system descri bed i n thi s work had several
advantages over the ol der cutaneous- rectal l ead system. The two
pri nci pal advantages were l ess muscul ar ri gi di ty and a hi gher percentage
of adequate semen sampl es. Possi bl y the di f f erent pre- anestheti c
tranqui l i zers used i n testi ng the two el ectroej acul ators may have had
some i nf l uence on the resul ts. The newer ej acul ator di d not provi de as
many adequate semen sampl es i n March- Apri l , 1981, as i n J anuary, 1982.
Thi s may have resul ted because March- Apri l i s l ate i n the coyote
breedi ng season (3) and some of the coyotes i n 1981 had been treated
wi th an anti f erti l i ty drug (busul f an).
The vol tage (18 Vrms) and wavef orm (1000 Hz si ne) used wi th the
two- ri ng probe worked wel l but may not have been opti mal si nce other
vol tages and wavef orms were not tested exhausti vel y, or tested at al l ,
unl ess an i ndi vi dual coyote f ai l ed to produce semen under standard
condi ti ons. Al though the saf ety and ef f i cacy of hi gh f requency current
f or el ectraej acul ati on were reported l ong ago (6), hi gh f requency has
sel dom been used, perhaps because the apparatus has been more expensi ve
than that needed f or 60 Hz.
The two- ri ng geometry was not necessari l y opti mal , but perf ormed
wel l i n the current study. Other two- ri ng rectal probes si mi l ar to the
probe used i n thi s study have been used i n ej acul ati on systems used f or
mi nk (7), rams (8), and f errets (9). The amount of exposed metal
surf ace on the f our- stri p rectal probe was obvi ousl y too great, al l owi ng
excessi ve current f l ow. Rectal probes wi th more than two ri ngs and wi th
varyi ng numbers of stri ps have reportedl y been used successf ul l y wi th
other carni vores such as the domesti c cat (l o), and wol ves, pol ar bears,
and l arge cats (11).
The vol ume of semen obtai ned wi th the two- ri ng probe i n thi s study
was si mi l ar to that reported previ ousl y f or coyotes when semen was
obtai ned wi th the ol der apparatus usi ng cutaneous- rectal l eads (2). No
reports of spermatozoa concentrati on i n coyote semen have been f ound,
58 JULY 1983 VOL. 20 NO. 1
THERIOGENOLOGY
but the val ue reported i n Tabl e 1 was consi derabl y l ess than that
reported f or the ti mber wol f (11) and wi thi n the range reported f or the
domesti c doq (12).
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
..I I
REFERENCES
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Kennel l y, J . J . Coyote reproducti on. I n: Coyotes. Bi ol oqy,
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Pl atz, C. C. and Seager, S. W. J . Semen col l ecti on by
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