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1051/parasite/1994014405

TRANSMISSION OF ANAPLASMA MARGINALE WITH ADULT BOOPHILUS MICROPLUS

TICKS FED AS NYMPHS ON CALVES WITH DIFFERENT LEVELS OF RICKETTSAEMIA

AGUIRRE D.H.*, G A I D O A.B.**, VINABAL A.E.*, D E ECHAIDE S.T.*** AND GUGLIELMONE A.A.***

Summary : Resume : TRANSMISSION DE ANAPLASMA MARGINALEPARBOOPHILUS

Nine splenectomised calves were infested with dissimilar numbers
of adult Boophilus microplus ticks 7 2 h after collection as
engorged nymphs from three non splenectomised calves with
different levels of Anaplasma marginale rickettsaemia. Successful
transmission of A. marginale appeared to be more dependent on
the level of rickettsaemia of the donor calves than on the number
of ticks attaching to the splenectomised calves, since infection was
transmitted only when the rickettsaemia was 0.3 % or greater.
Field transmission would thus depend on the rate of tick migration
amongst susceptible hosts and the rickettsaemia level of cattle on
which the ticks fed previously.
KEY WORDS : Anaplasma marginale, rickettsaemia level. Boophilus
microplus, transtadial transmission.
MOTS CLES : Anaplasma marginale, taux de rickettsémie. Boophilus
microplus, transmission transstadiale.
MICROPLUS(TIQUE-IXODIDÉS)RÉCOLTÉESAUSTADE DE, NYMPHES,SURDES
VEAUX PRÉSENTANT DIFFÉRENTS TAUX DE RICKETTSÉMIE
Neuf veaux splenectomisés ont été infestés par des Boophilus
microplus adultes récoltés 72 h auparavant au stade de nymphes,
gorgées sur trois veaux donneurs non splenectomisés présentant
différents taux de rickettsémie dues à Anaplasma marginale. Le
succès de la transmission de la rickettsiose dépend plus de la
rickettsémie des veaux donneurs que du nombre de tiques fixées sur
les veaux receveurs splénectomisés, puisque A. marginale n'était
transmis seulement lorsque les veaux donneurs présentaient des
rickettsémies égales ou supérieures à 0,3 %. La transmission
naturelle semble donc dépendre du flux de migration des
B. microplus entre hôtes réceptifs et du taux de rickettsémie des
bovins donneurs infestés.

INTRODUCTION o f B. microplus b e t w e e n cattle has b e e n d o c u m e n t e d

(Mason a n d Norval, 1 9 8 1 ) . H o w e v e r the proportion
o f adult B. microplus migrating amongst cattle under
A n a p l a s m a marginale is widespread in cattle
natural field conditions w a s recorded as 0.01 % over
of tropical and subtropical environments
a p e r i o d o f eight days in n o r t h w e s t e r n Argentina
infested with Boophilus microplus ticks, long
( G u g l i e l m o n e and Mangold, in press). Consequently
incriminated as one of its main biological vectors.
the infection rate o f B. microplus with A. marginale is
However, a study from the area of Argentina infested
p r e s u m e d to b e high in order to b e c o m e a relevant
with this boophilid showed no correlation between
transmitter o f this organism. T h e authors tested this
B. microplus infestation and A. marginale infection
hypothesis by transferring different numbers of
(Rios, Aguirre and Gaido, 1990). This fact might be
B. microplus ticks from calves with t h r e e different
expected since B. microplus is one-host-tick
levels o f A. marginale r i c k e t t s a e m i a to s u s c e p t i b l e
(Hoogstraal and Aeschlimann. 1982) and transovarial
s p l e n e c t o m i s e d calves.
transmission of A. marginale appears to be a rare
event (Uilenberg, 1968, 1970, 1973; Potgieter, 1979).
Therefore tick and rickttesial cycles are not MATERIALS AND METHODS

T
synchronised.
h e s t u d y w a s c a r r i e d o u t in t h e I n s t i t u t o
On the other hand, intra and transtadial transmission
of A. marginale by B. microplus have been described Nacional d e T e c n o l o g ï a A g r o p e c u a r i a , Salta,

(Connell and Hall, 1972; Connell, 1974, Thompson Argentina, using 2 - 3 - m o n t h s - o l d Holstein

and Roa, 1978) but the importance of this date under calves purchased from an area free o f B. microplus.
different field conditions remains unknown. Migration T h e y were negative for the p r e s e n c e o f antibodies to
A. marginale as determined b y the card agglutination
test (CAT) (Amerault a n d Roby, 1 9 6 8 ) . All calves were
* Instituto Nacional de Tecnología Agropecuaria, Estación
maintained tick a n d fly free in isolation b o x e s .
Experimental Agropecuaria Salta, CC 228, CP 4400 Salta, Argentina,
** Consejo Nacional de Investigaciones Científicas y Tecnológicas, T h e B. microplus c o l o n y n a m e d 188, free o f Babesia
Estación Experimental Agropecuaria Salta, CC 228, CP 4400 Salta, and Anaplasma infection a n d maintained o n calves
Argentina also free o f these organisms, a n d the cryopreserved
*** Instituto Nacional de Tecnología Agropecuaria, Estación
Experimental Agropecuaria Rafaela, CC 22. CP 2300 Rafaela (Santa S I P strain o f A. marginale with n o a p p e n d a g e s (Rios
Fe), Argentina. et al, 1 9 8 8 ) w e r e used throughout.
 AGUIRRE D.H.,GAIDOA.B., VINABAL A.E., DE ECHAIDE S.T. and GUGLIELMONE A.A.

Table I. - Transtadial transmission of Anaplasma marginale to splenectomised calves by Boophilus microplus adult ticks fed to nymphal
engorgement on infected calves with different rickettsaemia levels.

Three calves A, B and C, were successively inoculated adult ticks a t t a c h e d ( c a l f A 2 ) . H o w e v e r n i n e adult

i n t r a v e n o u s l y with 2 0 0 m i l l i o n SIP A. marginale ticks fed as nymphs on calf C with 0.02 %
organisms after thawing aliquots in water at 37°C. On rickettsaemia on the day o f nymphal collection w e r e
days 14 and 20 after A. marginale inoculation each unable to infect s p l e n e c t o m i s e d calf C2.
calf was infested with 10 000, 188 B. microplus larvae T h e m e a n pre-patent period (interval from the time o f
30-days-old. Fifteen days after the first infestation with adult tick attachment t o first detection of A.
ticks, 300 engorged nymphs were taken from each calf marginale in the b l o o d ) for the five s p l e n e c t o m i s e d
and conditioned in darkness at 27 ± 1°C, 83-86 % of c a l v e s w h i c h d e v e l o p e d infection w a s 2 6 . 8 ± 4 . 9 1
relative humidity for 72 h to allow them to moult days. T h e m i n i m u m haematocrit i n d e x varied from
through the adult stage. Adult ticks ( s e x ratio = 1) 0.11 to 0 . 1 4 a n d the m a x i m a l t e m p e r a t u r e ranged
were transferred to shaven areas, under cotton patches from 3 9 . 8 to 40.8°C.
on the n e c k , o f n i n e s p l e n e c t o m i s e d calves w h i c h
S p l e n e c t o m i s e d c a l v e s A2 a n d B 3 d i e d o f a c u t e
were divided into three groups. Three calves A l . A2 anaplasmosis without seroconverting, while calves B l
and A 3 ; B l , B2 and B 3 ; C l , C2 and C3 were infested and B2 had seroconverted by day 33 from infestation
with ticks from d o n o r calf A. B or C, respectively. and calf Al was found positive to CAT on day 7 0 .
Within each group, each of the three calves received a Splenectomised calves A3, C1, C2 and C3 remained
different n u m b e r o f ticks as detailed in table I, to uninfected as assessed by negative b l o o d smears and
obtain an estimation of the minimum number of ticks the persistent a b s e n c e of antibodies to A. marginale.
required to transmit S1P A. marginale.
T h e n u m b e r o f ticks (females and males) attaching,
rectal temperature and rickettsaemia by observation DISCUSSION
o f Giemsa stained thin smears prepared daily from all
calves were recorded. Haematocrit index using the Previous studies of transtadial transmission of
micro haematocrit t e c h n i q u e was evaluated three A. marginale b y B. microplus ticks were
times w e e k l y and the CAT was performed on days
c a r r i e d out with d o n o r c a l v e s in which
33, 70 and 100 after tick infestation.
rickettsaemias w e r e higher than 10 % on the day of
tick c o l l e c t i o n ( C o n n e l l a n d Hall, 1 9 7 2 ; Connell,
1 9 7 4 ; T h o m p s o n and Roa, 1978) and the n u m b e r of
RESULTS ticks attaching to the recipient calves used to test the
transmission o f infection w e r e not specified. O n the
he main results are presented in table I. T h e other hand Samish, Pipano and Hadani ( 1 9 9 3 ) , using
proportion o f B. microplus adults attaching to Boophilus annulatus, w e r e a b l e to transmit A.
the splenectomised calves varied from 0 % to marginale by transferring 220 e n g o r g e d n y m p h s from
100 %. All calves infested with ticks fed as nymphs a s p l e n e c t o m i s e d calf with 4 % rickettsaemia w h e n
on d o n o r calves with at least 0.3 % o f erythrocytes the ticks w e r e collected, to a n o t h e r s p l e n e c t o m i s e d
infected with A. marginale on the day o f nymphal calf, w h e r e o n l y t w o e n g o r g e d f e m a l e ticks w e r e
collection b e c a m e infected even when only three recovered ( n u m b e r o f males attached not stated).

Parasite, 1994, 1. 405-407

406 Note de recherche
 TRANSMISSION OF ANAPLASMAMARGINALEWITHADULT BOOPHILUS MICROPLUS TICKS FED AS NYMPHS ON CALVES WITH DIFFERENT LEVELS OF RIEKETTSAEMIA

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