Isolation, Tissue Distribution, and Molecular Characterization of Toxoplasma gondii from

Chickens in Grenada, West Indies

Author(s): J. P. Dubey, M. I. Bhaiyat, C. de Allie, C. N. L. Macpherson, R. N. Sharma, C.
Sreekumar, M. C. B. Vianna, S. K. Shen, O. C. H. Kwok, K. B. Miska, D. E. Hill and T. Lehmann
Source: The Journal of Parasitology, Vol. 91, No. 3 (Jun., 2005), pp. 557-560
Published by: Allen Press on behalf of American Society of Parasitologists
Stable URL: http://www.jstor.org/stable/20059714
Accessed: 27-12-2015 03:06 UTC

Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at http://www.jstor.org/page/
info/about/policies/terms.jsp

JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content
in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship.
For more information about JSTOR, please contact support@jstor.org.

Allen Press and American Society of Parasitologists are collaborating with JSTOR to digitize, preserve and extend access to
The Journal of Parasitology.

http://www.jstor.org

This content downloaded from 144.82.108.120 on Sun, 27 Dec 2015 03:06:05 UTC
All use subject to JSTOR Terms and Conditions
 ./. Parasitai, 91(3), 2005, pp. 557-560
? American Society of Parasitologists 2005

ISOLATION,TISSUE DISTRIBUTION,AND MOLECULAR CHARACTERIZATIONOF

TOXOPLASMA GONDII FROM CHICKENS INGRENADA, WEST INDIES
J. P. Dubey, M. I. Bhaiyat*, C. de Allie*, C. N. L. Macphersonf, R. N. Sharma , C. Sreekumar, M. C. B. Vianna,
S. K. Shen, O. C. H. Kwok, K. B. M ska,
i D. E. Hill, and T. Lehmann^
United States Department of Agriculture, Agricultural Research Service, Animal and Natural Resources Institute, Animal Parasitic Diseases
Laboratory, Building 1001, Beltsville, Maryland 20705-2350. e-mail: jdubey@anri.barc.usda.gov

abstract: The prevalence of Toxoplasma gondii in free-range chickens is a good indicator of the prevalence of T. gondii
oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 102 free-range chickens (Gallus
domesticus) from Grenada was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT).
Antibodies were found in 53 (52%) chickens with titers of 1:5 in 6, 1:10 in 4, 1:20 in 4, 1:40 in 4, 1:80 in 15, 1:160 in 9, 1:
320 in 5, 1:640 in 4, and 1:1,280 or greater in 2. Hearts, pectoral muscles, and brains of 43 seropositive chickens with MAT
titers of 1:20 or greater were bioassayed individually in mice. Tissues of each of 10 chickens with titers of 1:5 and 1:10 were
pooled and bioassayed in mice. Tissues from the remaining 49 seronegative chickens were pooled and fed to 4 T. gondii-free
cats. Feces of cats were examined for oocysts; they did not shed oocysts. T. gondii was isolated from 35 of 43 chickens with
MAT titers of 1:20 or greater; from the hearts, brains, and pectoral muscles of 2, hearts and brains of 20, from the hearts alone
of 11, and brains alone of 2. T. gondii was isolated from 1 of 10 chickens with titers of 1:5 or 1:10. All 36 T. gondii isolates
were avirulent for mice. Genotyping of these 36 isolates using polymorphisms at the SAG2 locus indicated that 29 were Type
III, 5 were Type I, 1 was Type II, and 1 had both Type I and Type III. Genetically, the isolates from Grenada were different
from those from the United States; Type II was the predominant type from the United States. Phenotypically, all isolates from
Grenada were avirulent for mice, whereas those from Brazil were mouse-virulent. This is the first report of isolation of T. gondii
from chickens from Grenada, West Indies.

Toxoplasma gondii infections are widely prevalent in human of the strains in their environment (Ruiz and Frenkel,
prevalent
beings and animals worldwide (Dubey and Beattie, 1988). Hu 1980).
mans become infected tissue from we found that 70% of 73 T. gondii isolates obtained
postnatally by ingesting cysts Recently,
undercooked meat, food or drink contaminated with from chickens from Brazil were
consuming asymptomatic free-range Type
oocysts, or by accidentally oocysts from the environ I (Dubey et al., 2002; Graham, da Silva et al., 2003;
ingesting Dubey,
ment. However, only a small percentage of adult hu Navarro et al., 2003), whereas from Egypt and
exposed Dubey, samples
mans develop clinical signs. It is unknown whether the severity the United States were dominated by either Type II or Type III,
of toxoplasmosis in immunocompetent persons is due to the but Type I was
absent Graham, Dahl, Hilali et al.,
(Dubey,
parasite strain, host or to other factors. 2003; Graham, Dahl, Sreekumar et al., 2003). II
variability, Dubey, Type
Toxoplasma gondii isolates have been classified into 3 ge isolates of T. gondii have not been found in chickens from
netic types (I, II, III) based on restriction Brazil. All 3 types were found in chickens from Argentina
fragment length poly
morphism (RFLP) (Howe and Sibley, 1995; Howe et al., 1997). (Dubey, Venturini et al., 2003). Nothing is known of the char
It has been suggested that Type I strains or recombinants of acteristics of isolates of T. gondii from animals or humans from

Types I and III are more likely to result in clinical ocular toxo Grenada, West Indies. In the present paper, we to
attempted
plasmosis (Howe et al., 1997; Fuentes et al., 2001; et al., isolate and genotype T. gondii from chickens from Grenada.
Grigg
2001; Boothroyd and Grigg, 2002; Aspinall et al., 2003; Ajzen Additionally, the distribution of T. gondii in the heart, brain,
berg et al., 2004), but
genetic characterization has been limited and pectoral muscles of chickens was compared.
essentially to isolates from patients ill with toxoplasmosis. Un
like these reports, Ajzenberg et al. (2002) found that most (73 MATERIALS AND METHODS
of 86) isolates from cases of congenital in hu
toxoplasmosis Naturally infected chickens
mans from France were II. Nothing is known of the ge
Type =
The chickens (n 102) came from 13 households from Grenada,
netic diversity of T. gondii isolates circulating in the general West Indies (Fig. 1). The properties were at least 500 m apart. They
human population. In animals, most isolates of T. gondii were were purchased, bled, and then killed by cervical dislocation in 4 batch
II or Type III, of clinical status and es of 25, 25, 27, and 25 in April and May 2004. Serum, heart, pectoral
Type irrespective (Howe
et muscle, and brain from each chicken were sent cold by air to Beltsville,
Sibley, 1995;
Mondragon al., 1998; Owen and Trees, 1999;
Maryland. Two to 3 days elapsed between the death of the chickens
Jungersen et al., 2002). T. gondii isolates differ in
markedly and when the samples arrived in Beltsville, The
Maryland. samples
their virulence to out-bred mice. Type I isolates are more vir were in excellent condition when they arrived.
ulent to mice than Types II and III. Because chickens become
infected from contaminated with oo Serological examination
mostly by feeding ground
of T. gondii in chickens is a good indicator Chicken serum
was tested for T. gondii
cysts, prevalence antibodies using 4 serum
dilutions (i.e., 1:5, 1:10, 1:20, and 1:200), with the modified aggluti
nation test (MAT) as described by Dubey and Desmonts (1987). After
Received 3 September 2004; revised 11 September 2004; accepted 30
the bioassays were completed, all positive chicken sera were run again
September 2004.
*
of Paraclinical School of Veterinary using 2-fold dilutions from 1:5 to 1:320.
Department Studies, Medicine,
St. George's University, Grenada, West Indies.
Bioassay for T. gondii infection
f Windward Island Research and Education Foundation, True Blue
Campus, St. George's University, Grenada, West Indies. All chicken tissues were for T. gondii infection. Brains,
bioassayed
$ Division of Parasitic Diseases, Centers for Disease Control and Pre pectoral muscles, and hearts of 43 chickens with MAT titers of 1:20 or
vention, 4770 Buford Highway, MS: F22, Chamblee, Georgia 30341. greater were each bioassayed in out-bred female Swiss
individually

557

This content downloaded from 144.82.108.120 on Sun, 27 Dec 2015 03:06:05 UTC
All use subject to JSTOR Terms and Conditions
558 THE JOURNAL OF PARASITOLOGY, VOL. 91, NO. 3, JUNE 2005

N RESULTS
-
O 8*u?wf8 -& to T. gondii were found in 53 of 102 chickens
f Antibodies
I J 9 Prospect $3 with titers of 1:5 in 6, 1:10 in 4, 1:20 in 4, 1:40 in 4, 1:80 in
Grenada '"^dS,??? ^ 15, 1:160 in 9, 1:320 in 5, 1:640 in 4, and 1:1,280 or greater
SIMark S? Patrick in 2 chickens. T. gondii was isolated from 36 seropositive

( chickens; from 1 with a titer of 1:10, from 2 with a titer of 1:

20, from 4 with a titer of 1:40, from 11 with a titer of 1:80,
from 9 with a titer of 1:160, from 3 with a titer of 1:320, from
St Andrew 4 with a titer of 1:640, and from both chickens with a titer of
SX John
1:1,280 or greater (Table I).
10 Toxoplasma gondii was isolated from 35 of 43 (82%) chick
UFitattc ens with MAT titers of 1:20 or more from the hearts, brains,
and pectoral muscles of 2, hearts and brains of 20, from the
hearts alone of 11, and from the brains alone of 2 (Table I). It
Mo?n<ro .?rand Etang
Point r\ National Park was also isolated from 1 chicken with a titer of 1:10. Four of
"9 Consiirtkw
Wgodt'ord >z ;^bt*at 5 mice inoculated with pooled tissues of chicken 95 were found
o^,***
StDavkJ to have in their brains. All 36 isolates were avirulent
-^t?
tissue
*?** cysts
(& for mice.
St Georgen A ':??
4 r -**#? Genotyping of 36 T. gondii isolates from chickens indicated
Grand Ans* Bay Upper Wobuni ^ Beaton ^ that 29 (80%) were Type III, 5 were Type I, 1was Type II and
Quaranbne ^
6 OM WcsterhaU 4 J 1 had both (see below) Type I and Type III (Table I). The
data were based on DNA from a single mouse in each
genetic
group of mice inoculated with chickens.

a??^ ^Lanc* aux ?pines Incomplete digestion was noticed after Sau 3A1 digestion of
Po<r4
the PCR products from the 5' terminal of the SAG2 gene from
the isolates from chicken 34. To determine the presence of
Figure 1. Map of Grenada showing 10 areas from where chickens the PCR were in
mixed genotypes, products directly sequenced
were obtained.
both directions and the chromatograms analyzed for the pres
ence of double The chromatogram of this isolate was
peaks.
Webster mice obtained from Taconic New York, found to have a small T peak along with a dominant C peak,
Farms, Germantown,
as described (Dubey et al., 2002). Each tissue was homogenized indi indicating the presence of both (GAT)T al?ele and (GAT)C al
vidually, digested in acidic pepsin, and washed, and the homogenate ?eles. Because there was no at the 3' end of the SAG2
digestion
was inoculated subcutaneously into 5 mice. In total, 15 mice were in
oculated with tissues of each chicken. Tissues from each of 10 chickens gene of this isolate (thus ruling out the presence of Type II), it
with titers of 1:5 or 1:10 were pooled and treated as described above,
was concluded that both genotypes III and I were present in the
but they were inoculated into 5 mice per chicken. isolate from chicken 34.
Tissues from 49 seronegative chickens were pooled in 4 batches of
16, 16, 8, and 9 chickens and fed separately to 4 T. gondii-free cats
(Dubey et al., 2002). Feces of cats were examined for shedding of T. DISCUSSION
gondii oocysts 3-14 days after ingesting chicken tissues as previously
described (Dubey, 1995). Fecal floats were incubated for 1 wk at room The threshold MAT titer that is indicative of T. gondii infec
temperature to allow oocyst sporulation and were bioassayed in mice tion in chickens has not been determined. Data se
comparing
(Dubey and Beattie, 1988). Tissue imprints of mice that died were ex of viable T. gondii from chickens are now
rology and recovery
amined for T. gondii tachyzoites or tissue cysts. Survivors were bled
accumulating (Dubey et al., 2002; Dubey, Graham, Dahl, Hilali
on days 39-42 postinoculation and a 1:25 dilution of serum from each
mouse was tested for T. gondii antibodies with the MAT. Mice were et al., 2003; Dubey, Graham, Dahl, Sreekumar et al., 2003;
killed 1-7 days after serological examination, and the brains of all mice Dubey, Graham, da Silva et al., 2003; da Silva et al., 2003).
were examined for tissue cysts as described (Dubey and Beattie, 1988).
Although T. gondii was isolated from a few chickens with MAT
The inoculated mice were considered infected with T. gondii when ta
titers of 1:5 or less, the likelihood of isolation increased with
chyzoites or tissue cysts were found in tissues.
MAT titer. The MAT employed in the current study is at present
Genotyping and sequencing the best test for detecting antibodies to T. gondii in chickens

Toxoplasma gondii DNA was extracted from the tissues of a single (Dubey et al., 1993). The lack of oocyst shedding by 4 cats that
infected mouse from each group, as described previously (Lehmann et consumed entire hearts, brains, and g of pectoral
20-25 muscles
al., 2000). The RFLP strain type of T. gondii isolates was determined of 49 chickens the of the MAT.
seronegative supports validity
by nested PCR on the SAG2 locus according to the method described
T. gondii was isolated from the hearts of 32, brains
Notably,
by Howe et al. (1997).
of 24, and pectoral muscles ofonly 2 chickens. In the present
The PCR products were sequenced from the 5' end of the SAG2 (to
the Type III genotype) chicken tissues were in an excellent state of preservation
determine gene from the isolate (Table I) from study,
chicken 34 (TgCkGr8). The amplicons were extracted from the gel and and thus an opportunity to compare tissue It
provided tropism.
sequenced in the forward and reverse directions using the Big Dye is clear from these studies that chicken hearts should be
always
terminator system, version 3.1 (Applied Biosystems, Foster City, Cali
included among tissues from chickens in attempts to isolate T.
fornia) using an ABI 3100 sequencer. The sequence chromatograms
were edited using Sequencher The amount of material did not affect the
4.1 software (Genecodes Corp., Ann Ar gondii. bioassayed
bor, Michigan). results because entire brains and hearts were bioassayed; ap

This content downloaded from 144.82.108.120 on Sun, 27 Dec 2015 03:06:05 UTC
All use subject to JSTOR Terms and Conditions
 DUBEY ET AL.?TOXOPLASMOSIS FROM CHICKENS INGRENADA 559

Table I. Isolation of Taxoplasma gondii from tissues of seropositive chickens from Grenada.

Isolation in mice* from chicken tissues

Chicken Houshold designation, Chicken Genotype
no. location Parish MAT titer Brain Heart Muscle (isolate designation)

1 Industry St. Mark 640 0 I (TgCkGrl)

3 Industry St. Mark 320 0 III (TgCkGr2)
5 Industry St. Mark 20 0 III (TgCkGr3)
16 Duquesne St. Mark 160 1 I (TgCkGr4)
26 L. Woburn St. George 160 0 III (TgCkGr5)
30 L. Woburn St. George 20 0 III (TgCkGr?)
31 L. Woburn St. George 160 0 III (TgCkGr7)
34 D, Upper Woburn St. George 1,280 0 I + III (TgCkGr8)
35 D, Upper Woburn St. George 40 0 III (TgCkGr9)
37 D, Upper Woburn St. George 160 4 I (TgCkGrlO)
47 E, Beaton St. David 80 0 III (TgCkGrl 1)
51 Old Westerhall St. David 640 0 III (TgCkGrl2)
52 Old Westerhall St. David 80 0 III (TgCkGrl3)
53 Old Westerhall St. David 320 0 III (TgCkGrl4)
54 Old Westerhall St. David 80 0 III (TgCkGrl5)
58 Old Westerhall St. David 160 0 III (TgCkGrl6)
59 Old Westerhall St. David 640 0 III (TgCkGrl7)
60 New Westerhall St. David 80 0 III (TgCkGrl8)
61 G, New Westerhall St. David 80 0 III (TgCkGrl9)
62 G, New Westerhall St. David 80 0 II (TgCkGr20)
64 G, New Westerhall St. David 320 0 III (TgCkGr21)
66 G, New Westerhall St. David 80 0 III (TgCkGr22)
67 G, New Westerhall St. David 80 0 III (TgCkGr23)
68 H, Prospect St. Patrick 1,280 0 III (TgCkGr24)
70 H, Prospect St. Patrick 40 0 I (TgCkGr25)
72 H, Prospect St. Patrick 640 0 I (TgCkGr26)
79 I,Woodford St. John 40 0 III (TgCkGr27)
82 I,Woodford St. John 80 0 III (TgCkGr28)
83 I,Woodford St. John 160 0 III (TgCkGr29)
84 I,Woodford St. John 80 0 III (TgCkGr30)
85 I,Woodford St. John 40 0 III (TgCkGr31)
86 I,Woodford St. John 160 0 III (TgCkGr32)
87 I,Woodford St. Andrew 160 0 III (TgCkGr33)
91 J, La Filette St. Andrew 160 0 III (TgCkGr34)
95 J, La Filette St. Andrew 10 f III (TgCkGr35)
97 J, La Filette St. Andrew 80 0 III (TgCkGr36)
* Five mice inoculated with each tissue.
t Heart, brain, and skeletal muscle were pooled; 4 of 5 mice were infected.

proximate weights of each brain, heart, and pectoral muscles chickens from the United States (Dubey, Graham, Dahl, Sreek

bioassayed were 3, 5, and 20 g, respectively. umar et al., 2003; Lehmann et al., 2003, 2004).
The mouse virulence data indicate that the isolates of T. gon The detection of viable T. gondii of 2 genotypes (Type I and
dii from Grenada are similar to isolates from Egypt, India, Mex Type III) genotypes in the tissues of a mouse inoculated with

ico, and the United States. None of the isolates from Egypt the tissues of chicken 34 is of interest. There has been only a

(Dubey, Graham, Dahl, Hilali et al., 2003), India (Sreekumar single report of isolation of viable T. gondii of 2 genotypes
et al., 2003), Mexico (Dubey, Morales et al., 2004), or the Unit from any host (Dubey, Graham, da Silva et al., 2003). The
ed States (Dubey, Graham, Dahl, Sreekumar et al., 2003; Leh authors reported the isolation of Type III and Type II genotypes
mann et al., 2003) was virulent for mice. The isolates from from the tissues of a chicken by mouse Of the 5 mice
bioassay.
Grenada were different from those from Argentina (Dubey, inoculated with the digested chicken tissues, 2 mice were found
Venturini et al., 2003), Brazil (Dubey et al., 2002; Dubey, Gra to be infected with Type I genotype, while the remaining 3 had
ham, da Silva et al., 2003; Dubey, Navarro et al., 2003), and Type III. In the present case, both genotypes were found in the
Peru (Dubey, Levy et al., 2004); isolates from these countries same mouse, thus leading to partial with Sau 3Al.
digestion
were virulent for mice. T. gondii isolates from chickens from However, sequence showed the presence of
chromatograms
Brazil were predominantly Type I, whereas they were predom both C and T peaks at the polymorphic site, confirming the

inantly Type III from Grenada; Type II was not found in Brazil presence of both al?eles. The efficacy of identifying multiple
and was rare in Grenada. I isolates were not found in genotype infections from depends on numerous factors,
Type samples

This content downloaded from 144.82.108.120 on Sun, 27 Dec 2015 03:06:05 UTC
All use subject to JSTOR Terms and Conditions
 560 THE JOURNAL OF PARASITOLOGY, VOL. 91, NO. 3, JUNE 2005

the most the relative rate of multiplication of Oliveira. 2003.

Toxoplasma gondii isolates from free ranging
important being
chickens from Rio
de Janeiro, Brazil: Mouse mortality, genotype,
different genotypes and number of passages (Ajzenberg et al.,
and oocyst shedding by cats. Journal of Parasitology 89: 851-853.
2002, 2004; Boothroyd and Grigg, 2002; Villena et al., 2004).-, M. Z. Levy, C. Sreekumar, O. C. H. Kwok, S. K. Shen, E.
The density of T. gondii in tissues of asymptomatic animals is Dahl, P. Thulliez, and T. Lehmann. 2004. Tissue distribution and
low. Thus, it is difficult to detect T. gondii from animal molecular characterization of chicken isolates of Toxoplasma gon
directly
dii from Peru. Journal of Parasitology 90: 1015-1018.
tissues. However, mixed genotypes have been detected in spec
-, E. S. Morales, and T. Lehmann. 2004. Isolation and genotyp
imens from patients with clinical toxoplasmosis (Fuentes et al.,
ing of Toxoplasma gondii from free-ranging chickens from Mexico.
2001; Aspinall et al., 2003). It is generally believed that Type Journal of Parasitology 90: 411-413.
I multiplies at a faster rate outgrowing less virulent genotypes -, I. T Navarro, D. H. Graham, E. Dahl, R. L. Freir?, L. B.
and thus may mask the other as passage Prudencio, C. Sreekumar, M. C. Vianna, and T. Lehmann. 2003.
genotypes progresses.
Characterization of Toxoplasma gondii isolates from free range
At present, there are no definite assays to diagnose the presence
chickens from Paran?, Brazil. Veterinary Parasitology 117: 229
of mixed genotype infections of T. gondii in animal tissues. 234.
The isolate in this case was genotyped from the original pas -, M. D. Ruff, M. E. Camargo, S. K. G. L. Wilkins, O.
Shen,
thus the chances of detection of mixed infec C. H. Kwok, and P. Thulliez. 1993. Serologie and parasitologic
sage, increasing
tion. responses of domestic chickens after oral inoculation with Toxo
plasma gondii oocysts. American Journal of Veterinary Research
54: 1668-1672.
ACKNOWLEDGMENTS -, M. C. Venturini, L. Venturini, M. Piscopo, D. H. Graham,
E. Dahl, C. Sreekumar, M. C. Vianna, and T Lehmann. 2003.
We thank Lilia Cabrera and K. Hopkins for their technical assistance.
Isolation and genotyping of Toxoplasma gondii from free-ranging
chickens from Argentina. Journal of Parasitology 89: 1063-1064.
LITERATURE
CITED Fuentes, I., J. M. Rubio, C. Ram?rez, and J. Alvar. 2001. Genotypic
characterization of Toxoplasma gondii strains associated with hu
Ajzenberg, D., A. L. Ba?uls, C. Su, A. Dum?tre, M. Demar, B. Car
man in Spain: Direct from clinical
toxoplasmosis analysis samples.
me, and M. L. Dard?. 2004. Genetic diversity, clonality and sex
Journal of Clinical Microbiology 39: 1566-1570.
uality in Toxoplasma gondii. International Journal for Parasitology
Grigg, M. E., J. Ganatra, J. C. Boothrooyd, and T. P. Margolis.
34: 1185-1196.
2001. Unusual abundance of atypical strains associated with human
-, N. Cogn?, L. Paris, M-H. P. Thulliez,
Besseres, D. Filli
ocular toxoplasmosis. Journal of Infectious Diseases 184: 633-639.
setti, H. Pelloux, P. Marty, and M.-L.
Dard?. 2002. Genotype
Howe, D. K., and L. D. Sibley. 1995. Toxoplasma gondii comprises
of 86 Toxoplasma gondii isolates associated with human congenital
three clonal lineages: Correlation of parasite genotype with human
toxoplasmosis, and correlation with clinical findings. The Journal
disease. Journal of Infectious Diseases 172: 1561-1566.
of Infectious Diseases 186: 684-689.
-, S. Honor?, F Derouin, L. D. Sibley. 1997. Determination of
Aspinall, T. V, E. C. Guy, K. E. Roberts, D. H. M. Joynson, J. E.
genotypes of Toxoplasma gondii strains isolated from patients with
Hyde, and P. F G. Sims. 2003. Molecular evidence for multiple
toxoplasmosis. Journal of Clinical Microbiology 35: 1411-1414.
Toxoplasma gondii infections in individual patients in England and
Jungersen, G., L. Jensen, M. R. Rask, and P. Lind. 2002. Non-lethal
Wales: Public health implications. International Journal for Para
infection parameters in mice separate sheep type II Toxoplasma
sitology 33: 97-103.
gondii isolates by virulence. Comparative Immunology, Microbi
Boothroyd, J. C, and M. E. Grigg. 2002. Population biology of Toxo
ology and Infectious Diseases 25: 187-195.
plasma gondii and its relevance to human infection: Do different and
Lehmann, T., C. R. Blackston, S. F Parmley, J. S. Remington,
strains cause different disease? Current Opinion in Microbiology J. P. Dubey. 2000. Strain typing of Toxoplasma gondii: Comparison
5: 438-442. of antigen-coding and house-keeping genes. Journal of Parasitology
da Silva, D. S. S., L. M. G. Bahia-Oliveira, S. K. Shen, O. C. H. 86: 960-971.
Kwok, T Lehmann, and J. R Dubey. 2003. Prevalence of Toxo -, D. H. Graham, E. Dahl, L. M. G. Bahia-Oliveira, S. M.
plasma gondii in chickens from an area in Southern Brazil highly and J. P. Dubey. 2004. Variation in the structure of Toxo
Gennari,
endemic to humans. Journal of Parasitology 89: 394-396. and the roles of selfing, drift, and epistatic selection
plasma gondii
Dubey, J. P. 1995. Duration of immunity to shedding of Toxoplasma in maintaining Genetics and Evo
linkage disequilibria. Infection,
gondii oocysts by cats. Journal of Parasitology 81: 410-415. lution 4: 107-114.
-, and C. P. Beattie. 1988. Toxoplasmosis of animals and man. -,-,-, C. Sreekumar, F. Launer, J. L. Corn, H. R.
CRC Press, Boca Raton, Florida, 220 p. and J. P. Dubey. 2003. Transmission of Toxo
Gamble, dynamics
-, and G. Desmonts. 1987. Serological responses of equids fed on a pig farm. Infection, Genetics and Evolution 3:
plasma gondii
Toxoplasma gondii oocysts. Equine Veterinary Journal 19: 337 135-141.
339. MONDRAGON, R., D. K. HOWE, J. P. DUBEY, AND L. D. SlBLEY. 1998.
-, D. H. Graham, C. R. Blackston, T Lehmann, S. M. Gennari, of Toxoplasma isolates from pigs. Jour
Genotypic analysis gondii
A. M. A. Ragozo, S. M. Nishi, S. K. Shen, O. C. H. Kwok, D. E. nal of Parasitology 84: 639-641.
Hill, and P. Thulliez. 2002. Biological and genetic characterisa Owen, M. R., and A. J. Trees. 1999. Genotyping of Toxoplasma gondii
tion of Toxoplasma gondii isolates from chickens (Gallus domes associated with abortion in sheep. Journal of Parasitology 85: 382
ticus) from S?o Paulo, Brazil: Unexpected findings. International 384.
Journal for Parasitology 32: 99-105. Ruiz, A., and J. K. Frenkel. 1980. Intermediate and transport hosts of
-, -, E. Dahl, C. Sreekumar, T. Lehmann, M. F. Davis, Toxoplasma gondii in Costa Rica. American Journal of Tropical
and T Y. Morishita. 2003. Toxoplasma gondii isolates from free Medicine and Hygiene 29: 1161-1166.
ranging chickens from the United States. Journal of Parasitology Sreekumar, C, D. H. Graham, E. Dahl, T. Lehmann, M. Raman, D.
89: 1060-1062. P. Bhalerao, M. C. B. Vianna, and J. P. Dubey. 2003. Genotyping
-,-,-, M. Hilali, A. El-Ghaysh, C. Sreekumar, O. of Toxoplasma gondii isolates from chickens from India. Veterinary
C. H. Kwok, S. K. Shen, and T Lehmann. 2003. Isolation and Parasitology 118: 187-194.
molecular characterization of Toxoplasma gondii from chickens Villena, L, M. Marle, M. L. Dard?, and J. M. Pinon. 2004. Toxo
and ducks from Egypt. Veterinary Parasitology 114: 89-95. plasma strain type and human disease: Risk of bias during parasite
-, -, D. S. da Silva, T. Lehmann, and L. M. G. Bah?a isolation? Trends in Parasitology 20: 160-162.

This content downloaded from 144.82.108.120 on Sun, 27 Dec 2015 03:06:05 UTC
All use subject to JSTOR Terms and Conditions