Search

Objectives: Establishment of a mouse model for congenital toxoplasmosis based on oral infection with oocysts from Toxoplasma gondii ME49 and its application for investigating chemotherapeutic options against congenital toxoplasmosis., Methods: CD1 mice were mated, orally infected with 5, 25, 100, 500 or 2000 oocysts and monitored for clinical signs and survival of dams and pups until 4 weeks post partum . The parasite burden in infected mice was quantified by real-time PCR in lungs, brains and, in the case of surviving pups, also in eyes. Seroconversion was assessed by ELISA. T. gondii cysts in brain were identified by immunofluorescence. In a second experiment, pregnant CD1 mice challenged with 20 oocysts/mouse were treated with buparvaquone or the calcium-dependent protein kinase 1 inhibitor bumped kinase inhibitor (BKI)-1294 and the outcome of infection was analysed., Results: T. gondii DNA was detected in the brain of all infected animals, irrespective of the infection dose. Seroconversion occurred at 3 weeks post-infection. Most pups born to infected dams died within 1 week post partum , but a small fraction survived until the end of the experiment. T. gondii DNA was detected in the brain of all survivors and half of them exhibited ocular infection. Chemotherapy with both compounds led to dramatically increased numbers of surviving pups and reduced cerebral infection. Most efficient were treatments with BKI-1294, with 100% survivors and only 7% brain-positive pups., Conclusions: BKI-1294 and buparvaquone exert excellent activities against transplacental transmission in pregnant mice., (© The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Ten pregnant sows were experimentally inoculated per os with T. gondii in order to investigate vertical and galactogenic transmission of the parasite and the evolution and maturation of the specific IgG humoral response in the sows and piglets. Five seronegative sows received 10 4 T. gondii (CZ isolate clone H3) sporulated oocysts during late-pregnancy (Exp. 1), three sows received 10 4 oocysts during mid-pregnancy (Exp. 2) and three sows from Exp. 1 (and two seronegative sows) were re-inoculated with 10 5 oocysts during a further pregnancy (late-pregnancy) (Exp. 3). Besides, six 4.5 week-old piglets inoculated per os with 5×10 3 oocysts were also included in the serological investigations. All animals seroconverted (PrioCHECK Toxoplasma Ab porcine ELISA, Prionics, Switzerland) by 2-3 weeks post inoculation (wpi) and remained seropositive for at least 38 weeks or until euthanasia. Four chronically infected sows from Exp. 1 and 2 were serologically monitored during a further pregnancy and no reactivation, but a decrease of the antibody levels was observed at farrowing (Exp. 4). In all experiments, the specific IgG-avidity was initially low, increased during the course of infection and after re-inoculations. An avidity index (AI) ≥40% could be used to rule out recent infections (<8 weeks) in most (15 of 16) animals. In some piglets (18.6% of 70) delivered by inoculated sows (Exp. 1 and 2), maternal antibodies were still detectable at 2 months (but not by 3 months) of age, with constant high avidity values, comparable to those of the dams at farrowing. In all experiments, the sows remained asymptomatic and delivered non-infected offspring at term. A total of 208 normal and 5 stillborn piglets delivered by the inoculated sows (Exp. 1-4) tested serologically negative before colostrum uptake. Placentas (n=88) from all sows and tissues (brain, liver, lung, heart, and masseter muscle) from 56 delivered piglets were analysed histopathologically and by real-time PCR for T. gondii with negative results. Colostrum and milk samples from all sows were negative by real-time PCR for T. gondii DNA. In addition, no seroconversion was observed in 16 piglets from seronegative dams that were transferred to infected dams one day after birth to detect a possible infection through colostrum or milk during the suckling period. Although vertical transmission of T. gondii was demonstrated in naturally infected pigs, many factors involved in the outcome of vertical transmission and congenital toxoplasmosis in pigs are still unknown., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Given the fact that numerous microbial species can be detected in pregnant female dogs, the objective of this study was to assess the transplacental transmission of Brucella canis, Ehrlichia canis, Neospora caninum and Toxoplasma gondii in stillborn puppies. This study involved 41 stillborn puppies, 78.6% of which were positive for T. gondii, 52.4% for N. caninum and 59.5% for B. canis. E. canis was not detected in any of the analyzed puppies. Pregnancy is an important physiological condition for the transmission of infectious agents to puppies and transplacental transmission may be epidemiologically relevant in the spread of these opportunistic agents., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The present study aimed to evaluate the transmission of toxoplasmosis (vertical and venereal) and its influence on milk production and reproductive problems of Lacaune sheep seropositives for Toxoplasma gondii. Males and females were serologically selected using indirect immunofluorescence method in three steps of the study. Step 1: In order to evaluate the influence of toxoplasmosis on milk production, the volume of milk produced by 40 sheep (22 seronegatives and 18 seropositives for T. gondii) was weekly measured throughout the lactation period. There were no significant differences between these two groups; in other words, toxoplasmosis did not affect milk production. Step 2: In order to assess T. gondii venereal transmission, five samples of semen from seropositive rams (n = 5) were tested by endpoint and real time PCR with two days of interval; however, these semen samples were PCR negatives for T. gondii. Step 3: To evaluate reproductive problems, 12 seropositive animals out of a flock of 68 pregnant ewes showed signs of reproductive problems, such as abortion or fetal resorption. T. gondii transplacental transmission was evaluated on blood drawn from newborn lambs (n = 41), and their respective seropositive mothers (n = 30) after single, double or triple births. Serological tests showed that 65.8% of the lambs had antibodies against this protozoan, indicating a high transmission from ewe to fetus during pregnancy. Therefore, it is concluded that toxoplasmosis in sheep may impair reproduction with a high percentage of vertical transmission., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Congenital transmission of Toxoplasma gondii may occur if the mother gets infected for the first time while pregnant. The risk of mother-to-child transmission depends on the gestation trimester at infection, being lowest in the first and highest in the last. Conversely, fetal damage is frequent and more severe at the beginning of pregnancy. The objective of this study was to evaluate congenital transmission and pathological aspects in the placenta and the fetus using a mouse model of congenital infection of the second gestation third. Forty-five female BALB/c mice were infected intravenously with 2.5-10.0 × 10(6) tachyzoites of the ME49 strain at middle gestation. Samples of maternal spleen and fetal/placental units were taken 72 h later. We determined parasite load and vertical transmission by qPCR, as well as damage macroscopically and by histopathology. With the lowest dose, 18% of the fetuses were infected. Also, 40% of fetuses/litter were altered, while this value was 10% in the control group (P < 0.05). These results are similar to those described in humans in terms of vertical transmission and fetal damage during the second third of gestation. The maternal spleen had 10-1000 times more tachyzoites than the placenta, and the later retained 90-99% of the parasites that could reach the fetus. Nevertheless, we found resorptions, abortions or fetal tissue damage in the presence but also in the absence of parasites. Our data indicate a strong protective effect of maternal organs and the placenta against fetal infection, but extensive damage of the later may led to resorption or abortion without vertical transmission., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Toxoplasmosis, caused by an obligate intracellular protozoan parasite, Toxoplasma gondii, is an worldwide parasitic disease, with significant importance for animal production and considerable impact to the public health. This study was aimed to evaluate the dynamic of the distribution of T.gondii in tissues of female Wistar rats and their puppies tissues, after the immunization by oral rote with irradiated tachyzoites. One week after pregnancy confirmation, rats was challenged by gavage with T. gondii bradyzoites, oocysts or tachyzoites of T. gondii. Forty-eight pregnant rats were grouped as follow: immunized and challenged with bradyzoites (BZ*); non-immunized and challenged with bradyzoites (BZ); immunized and challenged with oocysts (OC*); non-immunized and challenged with oocysts (OC); immunized and challenged with tachyzoites (TZ*); non-immunized and challenged with tachyzoites (TZ); only immunized (I); control group (C). After parturition the rats were sacrificed and the tissues were researched for the DNA of T. gondii by polymerase chain reaction (PCR) and the parasite load determined by the quantitative PCR (qPCR). It was verified that the immunization with irradiated tachyzoites of T. gondii induced the reduction of parasitic load in most organs analyzed, although not prevent the establishment of infection with the parasite. And also, the immunization showed a favorable effect on the birth rate and litter size., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Parasites with complex life cycles are expected to manipulate the behaviour of their intermediate hosts (IHs), which increase their predation rate and facilitate the transmission to definitive hosts (DHs). This ability, however, is a double-edged sword when the parasite can also be transmitted vertically in the IH. In this situation, as the manipulation of the IH behaviour increases the IH death rate, it conflicts with vertical transmission, which requires healthy and reproducing IHs. The protozoan Toxoplasma gondii, a widespread pathogen, combines both trophic and vertical transmission strategies. Is parasite manipulation of host behaviour still adaptive in this situation? We model the evolution of the IH manipulation by T. gondii to study the conflict between these two routes of transmission under different epidemiological situations. Model outputs show that manipulation is particularly advantageous for virulent strains and in epidemic situations, and that different levels of manipulation may evolve depending on the sex of the IH and the transmission routes considered. These results may help to understand the variability of strain characteristics encountered for T. gondii and may extend to other trophically transmitted parasites.

It is widely accepted that behavioural changes induced by Toxoplasma gondii are an adaptation of the parasite to enhance transmission to its cat definitive host. In our opinion, this explanation requires a rethink. We argue that the experimental evidence that observed behavioural changes will enhance transmission to cats is not convincing. We also argue that cats and sexual reproduction may not be essential for transmission and maintenance of this parasite. Thus, the selection pressure to infect a cat may not be sufficiently strong for the evolution of adaptive host manipulation to have occurred in order to enhance predation by cats., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Six ovine fetal brains were harvested 33 to 35 days postchallenge from 5 ewes, each of which was given 3000 Toxoplasma gondii oocysts on day 90 of pregnancy. Histopathologic examination of transverse sections taken at 13 levels in the fetal brains revealed the presence of toxoplasmosis-related lesions in all 6 brains. However, lesions were not randomly distributed (P = .007); they were most numerous at the level of the optic tract, the rostral margin of the pons, and 4 mm caudal to the ansate sulcus and were absent in all sections at the level of the caudal cerebellum. Lesion distribution may be due to hemodynamic factors, differences in the expression of endothelial surface receptor molecules at the level of the blood-brain barrier, or the presence of localized permissive/inhibitory factors within the brain. The results have implications for the selection of areas of brain from aborted ovine fetuses to be examined histopathologically for laboratory diagnosis.

Almost uniform protection against congenital toxoplasmosis initiated by inoculations with cysts and oocysts of the parasite was seen in the hamster model, among strains of different genotypes. Because the RH immunization prior to pregnancy has to be controlled with medication for most of the hamsters to survive, and also some congenital transmission of Toxoplasma was observed during the chronic stage of the infection, the hamster is considered less practical than the rat and the BALB/c mouse models. It is concluded that the hamster model closely resembles protection against congenital infection in nature, where most of the pregnant women and ewes that experienced a toxoplasma infection previously, protect their fetuses against an infection with the parasite during pregnancy., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The protozoan parasite Toxoplasma gondii is prevalent worldwide and can infect a remarkably wide range of hosts despite felids being the only definitive host. As cats play a major role in transmission to secondary mammalian hosts, the interaction between cats and these hosts should be a major factor determining final prevalence in the secondary host. This study investigates the prevalence of T. gondii in a natural population of Apodemus sylvaticus collected from an area with low cat density (<2·5 cats/km2). A surprisingly high prevalence of 40·78% (95% CI: 34·07%-47·79%) was observed despite this. A comparable level of prevalence was observed in a previously published study using the same approaches where a prevalence of 59% (95% CI: 50·13%-67·87%) was observed in a natural population of Mus domesticus from an area with high cat density (>500 cats/km2). Detection of infected foetuses from pregnant dams in both populations suggests that congenital transmission may enable persistence of infection in the absence of cats. The prevalences of the related parasite, Neospora caninum were found to be low in both populations (A. sylvaticus: 3·39% (95% CI: 0·12%-6·66%); M. domesticus: 3·08% (95% CI: 0·11%-6·05%)). These results suggest that cat density may have a lower than expected effect on final prevalence in these ecosystems.

We compared the transplacental-transmission ability of Toxoplasma gondii and Neospora caninum in cattle. One uninfected pregnant heifer served as control, while three were inoculated with N. caninum K9WA strain and four with T. gondii RH strain at their midgestational period. Both infected groups showed clinical signs and antibodies either to N. caninum or T. gondii, while the control animal was normal. Two (50%) Toxoplasma dams aborted on days 6 and 11 postinoculation. T. gondii tachyzoites were found in various organs of those dams that had abortions but not in their fetuses. Two Neospora dams did not abort but gave birth to subclinically infected calves. The remaining two Toxoplasma dams and one from Neospora group became recumbent. Those two dams and their fetuses showed disseminated Toxoplasma DNA, but no Neospora DNA was found. Our findings suggest that maternal toxoplasmosis could be a cause of abortion and congenital toxoplasmosis in cattle, especially when they are infected by virulent strains.

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes a variety of clinical syndromes, but the infection is severe in immunocompromised individuals and during pregnancy due to the possibility of transplacental transmission of the parasite causing congenital toxoplasmosis. Vertical transmission of the parasite usually occurs when females are primarily infected during pregnancy. Calomys callosus is resistant to T. gondii ME49 strain, which presents a moderate virulence and congenital disease occurs only during the acute phase of infection. The aim of this study was to determine whether vertical transmission occurs when females of C. callosus chronically infected with ME49 strain of T. gondii are reinfected with a highly virulent strain (RH, type I). Females were infected with cysts of the ME49 strain. On the 1st day of pregnancy, animals were reinfected with tachyzoites of the RH strain. In the 19th day of pregnancy, placentas and embryos were processed for morphological analysis, immunohistochemistry and for detection of the parasite by PCR and mouse bioassay. Morphological and immunohistochemical analyses revealed the presence of parasites only in placental tissues. Mouse bioassay results showed seroconversion only in mice that were inoculated with placental tissues. Also, T. gondii DNA was detected only in placental samples. Congenital toxoplasmosis does not occur in C. callosus females chronically infected with the moderately virulent ME49 strain of T. gondii and reinfected with the highly virulent RH strain, thus indicating that primary T. gondii infection before pregnancy leads to an effective long-term immunity preventing transplacental transmission to the fetus., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Congenital toxoplasmosis can cause fetal damage in humans and domestic animals. This study was focused on the effects of Toxoplasma gondii (Prugniaud strain) infection at different stages of pregnancy on the offspring of mice. Results showed that newborn mice from all infected groups were significantly lower in weight than those from the control group but significant difference was not found among these groups at day 60 after birth. The survival rate of the offspring from the group of mice infected at the earlier stage of pregnancy was significantly lower than those of infected and control groups. The positive offspring (with cysts found in their brain tissues) born from the mice infected at the earlier and intermediate stages of pregnancy showed a shorter latency and greater number of errors in the step-through passive avoidance test than those born from the mice infected at the late stage of pregnancy, the control group and the negative offspring from the infected groups. The number of cysts in the brain tissue was significantly higher in the offspring born from the groups of mice infected at the earlier and intermediate stages of pregnancy than those from the group of mice infected at the late stage of pregnancy. In addition, our results indicated that a high congenital transmission rate (90%) occurred in this NIH mouse model. In conclusion, the earlier and intermediate maternal infection of T. gondii can result in severe congenital toxoplasmosis, exhibiting conditions such as stillbirth or non-viability, and learning or memory capability damage in this mouse model. These results not only provide useful data for better understanding the effects of T. gondii infection on the offspring of mice infected at different stages of pregnancy but also for better consideration of the effect of this infection on other mammalian hosts including humans., (Copyright © 2010 Elsevier Inc. All rights reserved.)

To investigate how different routes of Toxoplasma gondii transmission influence the antibody response and infection status of deer mice (Peromyscus maniculatus), 80 mice were orally infected with 1, 5, 10, or 100 T. gondii oocysts. Ten weeks postinfection, 15 T. gondii -seropositive female mice were bred and allowed to produce 2 litters. Evidence of persistent T .gondii infection in orally infected mice was detected by serology and DNA amplification in mice from all 4 oocyst treatment groups, including those that received only a single T. gondii oocyst. Congenital transmission of T. gondii was detected by polymerase chain reaction (PCR) in 7/8 first and 4/7 second litters. Toxoplasma gondii was also detected by PCR in 9/30 congenitally infected offspring 16 wk after birth, despite the fact that detectable serological titers had waned. These findings raise questions about the applicability of serological testing to assess the prevalence of T. gondii infection in deer mice and other rodents in the wild. Additionally, the detection of frequent congenital transmission suggests that deer mice could help maintain T. gondii in the environment even in the absence of definitive feline hosts.

Toxoplasmosis is a parasitic zoonosis having worldwide distribution; it infects many human and animal populations and is produced by the parasite Toxoplasma gondii, this being of great importance in contagion of pregnant women since this zoonosis causes illness in the fetus through transplacental infection. A constant mother-foetus infection rate has traditionally been used. Nevertheless, there is evidence of a strong relationship between a mother's gestation week and the moment when a foetus becomes infected. This work describes congenital toxoplasmosis transmission dynamics by using an age-structured model taking a mother's gestational week into account. The model was adapted to a space-time model describing T. gondii dispersion through cats; this provided the amount of parasites in the environment depending on which a pregnant woman would become infected. Simulations were done, varying the amount of parasites and the pregnant mother-inoculum distance. Some parameters related to possible control measures were also varied. Important changes were found when comparing infected pregnant female population patterns, when a constant mother-fetus infection rate was used and when a dependent pregnancy week rate was used. This led to concluding that the gestation week in which the mothers became infected is fundamental in infection being transmitted to their fetuses.

Toxoplasma gondii is a highly ubiquitous and prevalent parasite. Despite the cat being the only definitive host, it is found in almost all geographical areas and warm blooded animals. Three routes of transmission are recognised: ingestion of oocysts shed by the cat, carnivory and congenital transmission. In natural populations, it is difficult to establish the relative importance of these routes. This paper reviews recent work in our laboratory which suggests that congenital transmission may be much more important than previously thought. Using PCR detection of the parasite, studies in sheep show that congenital transmission may occur in as many as 66% of pregnancies. Furthermore, in families of sheep on the same farm, exposed to the same sources of oocysts, significant divergent prevalences of Toxoplasma infection and abortion are found between different families. The data suggest that breeding from infected ewes increases the risk of subsequent abortion and infection in lambs. Congenital transmission rates in a natural population of mice were found to be 75%. Interestingly, congenital transmission rates in humans were measured at 19.8%. The results presented in these studies differ from those of other published studies and suggest that vertical transmission may be much more important than previously thought.

Toxoplasma gondii infection is common worldwide and highly important to pregnant women as it can be transmitted to the fetus via the placenta. This study aimed at evaluating the prevention of placental transmission in two different strains after chronic infection with each one of the strains. A BALB/c mice model was inoculated 30days before breeding (immunization) and re-infected 12 and 15days after pregnancy (challenge). Seven experimental groups were assayed: G1: ME49-immunization (type II), M7741-challenge (type III); G2: M7741-immunization, ME49-challenge; G3, ME49-immunization; G4: M7741-immunization; G5: ME49-challenge; G6: M7741-challenge; G7: saline solution inoculation. Serology, mouse bioassay, PCR and RLFP of the uterus, placenta and fetus were performed to determine the congenital transmission of the strains challenged after chronic infection. IgG T. gondii antibodies were detected in G1, G2, G3 and G4, but not in G5, G6 and G7. All animals of G5 and G6 were IgM-positive. Congenital infection was not detected by bioassay and PCR. Nonetheless, placentas from G3 and G4 resulted positive but no corresponding fetal infection was detected. G1 and G2 did not show the genotype of the strain challenged during pregnancy, only those of chronic infection. Thus, the chronically infected BALB/c mice showed no re-infection after inoculation with another strain during pregnancy. Further studies with different parasite loads and different mice lineages are needed.

To date, little is known about the dynamics of vertical transmission of Toxoplasma gondii in Australian marsupials. Studies in mice demonstrate that vertical transmission of T. gondii is common and that chronically infected mice can transmit T. gondii to successive generations. In this study, PCR and immunohistochemistry were used to detect T. gondii in chronically infected marsupial dams and their offspring. T. gondii was detected in the unfurred pouch young of 2 out of 10 chronically infected western grey kangaroos (Macropus fuliginosus) and in the unfurred pouch young of a brush-tailed bettong (Bettongia penicillata). Results of the study suggest that vertical transmission of T. gondii can occur in chronically infected Australian marsupials.

The objective of the research was to test the hamster for a model of transmission of congenital toxoplasmosis. A non-invasive method for the diagnosis of pregnancy in hamsters was designed, with a specificity and a sensitivity of 70.2 and 94.7%, respectively (n=168). Of 32 females with a chronic toxoplasma infection, 3 transmitted Toxoplasma congenitally during their first pregnancy, but not during the subsequent pregnancy. Congenital transmission rates of infections initiated during pregnancy with 2 stages of 2 strains of Toxoplasma were in the range of 33 to 100% of the 76 females inoculated. Only 1 of 17 females transmitted the parasite exclusively via milk. It was concluded that the hamster is a promising species for a model of transmission of congenital toxoplasmosis.

The objective of this study was to refine the rat model of congenital toxoplasmosis. In Fischer rats we found that visualization of spermatozoa in vaginal exudates and the detection of at least 6g body weight increase between days 9 and 12 of pregnancy, allowed the diagnosis and timing of pregnancy with 60% specificity and 84% sensitivity. A dose of 10(4) Toxoplasma gondii bradyzoites or 10(2) T. gondii oocysts of the Prugniaud strain resulted in more than 50% of congenital infection of the rat litters. Transmission of T. gondii via lactation was not detected in rats inoculated with either bradyzoites or oocysts. Bioassays of 51 neonates born from mothers inoculated with bradyzoites (in tissue cysts) and 29 neonates from mothers inoculated with oocysts demonstrated that both liver and lungs can be used for the diagnosis of congenital transmission in this model.

Clinical toxoplasmosis is most severe in congenitally-infected hosts. In humans, transmission of Toxoplasma gondii from the mother to the foetus is considered to be most efficient during the last trimester of pregnancy but clinical congenital toxoplasmosis is more severe if transmission occurs during the first trimester. However, there are no data on the rate of congenital transmission of T. gondii with respect to gestational age in any host during natural infection. In the present study, attempts were made to isolate T. gondii by bioassay in mice inoculated with tissues from foetuses of 88 naturally-exposed white-tailed deer from Iowa and Minnesota. Viable T. gondii was isolated from foetuses of six of 61 deer in early pregnancy (45-85 days of gestation) from Iowa and foetuses of nine of 27 deer from Minnesota in mid-gestation (130-150 days) of a gestational period of 7 months. The 15 T. gondii isolates obtained from foetal deer were PCR-restriction fragment length polymorphism genotyped using polymorphisms at 10 nuclear markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and an apicoplast marker, Apico. Five genotypes were revealed, including the clonal Type II and III lineages, and three non-clonal genotypes. DNA sequencing analysis of representative isolates at loci SAG2, c22-8, L358 and PK1 revealed that the three non-clonal genotypes are closely related to the clonal Type I, II and III lineages. It is very likely that these non-clonal genotypes were derived from genetic crosses among the three clonal Type I, II and III lineages. The most common genotype was Type II, commonly found in humans in North America and Europe, suggesting the possible link of transmission from game animals to humans.

Toxoplasma gondii is a protozoan parasite capable of infecting almost all warm-blooded animals. The cat is the definitive host and becomes infected by consuming contaminated meat or infected prey. Humans can act as intermediate hosts and in healthy individuals the infection is mild and self-limiting. In pregnant women it can cause spontaneous abortions and foetal abnormalities and is capable of inducing serious illness in immuno-compromised patients. In infested dwellings, mice could act as intermediate hosts and play a role in the persistence/propagation of the disease. A total of 200 mice were trapped alive in 27 infested properties in Manchester, UK, and screened for Toxoplasma infection; 59% tested positive. Evidence of vertical transmission from infected dams to foetus was found, possibly maintaining the infection in urban areas. These findings have important implications when considering approaches to rodent control.

Toxoplasma gondii is one of the few pathogens that can cross the placenta. Frequency and severity of transmission vary with gestational age. While acquired toxoplasmosis is already well explored, the control of maternal-foetal transmission of the parasite remains almost unknown. This is partly due to inherent inadequacies of animal models. This review summarises the studies which have been undertaken and shows that the mouse is a valuable model despite obvious differences to the human case. The paramount role of the cellular immune response during primary infection has been consistently shown. Surprisingly, IFN-g has a dual role in this process. While its beneficial effects in the control of toxoplasmosis are well known, it also seems to have transmission-enhancing effects within the placenta and can also directly harm the developing foetus. This shows the importance of designing vaccines which protects both mother and foetus. Therefore, it is useful to study the mechanisms of natural resistance against transmission during a secondary infection. In this setting, the process is more complicated, involving cellular, but also humoral components of the immune system. In summary, even if the whole process is far from being elucidated, important insights have been gained so far which will help us to undertake rational vaccine research.

In September 2004, a neonatal sea otter pup was found alive on the beach in northern Monterey Bay, CA. Efforts to locate the mother were unsuccessful. Due to a poor prognosis for successful rehabilitation, the pup was euthanized. Postmortem examination revealed emaciation, systemic lymphadenopathy and a malformation of the left cerebral temporal lobe. On histopathology, free tachyzoites and tissue cysts compatible with Toxoplasma gondii were observed in the brain, heart, thymus, liver, lymph nodes and peri-umbilical adipose. The presence of T. gondii within host tissues was associated with lymphoplasmacytic inflammation and tissue necrosis. Immunofluorescent antibody tests using postmortem serum were positive for anti-T. gondii IgM and IgG (at 1:320 and 1:1280 serum dilution, respectively), but were negative for IgG directed against Sarcocystis neurona and Neospora caninum (<1:40 each). Brain immunohistochemistry revealed positive staining for tachyzoites and tissue cysts using antiserum raised to T. gondii, but not S. neurona or N. caninum. T. gondii parasite DNA was obtained from extracts of brain and muscle by PCR amplification using the diagnostic B1 locus. Restriction enzyme digestion followed by gel electrophoresis and DNA sequencing confirmed the presence of Type X T. gondii, the strain identified in the majority of southern sea otter infections.

Toxoplasma gondii is a ubiquitous parasite with a widespread distribution both in terms of geographical and host range. Although the definitive host is the cat, it is also a major health hazard to domestic animals and humans. Three routes of transmission are recognised (infection from the cat, carnivory and congenital transmission). We aimed to assess the relative importance of congenital transmission, using sheep as a model system, due to the lack of carnivory. We report, using PCR as a diagnostic tool, that congenital transmission occurs with high frequency (69%). If transmission from oocysts was important in sheep, we would expect sheep reared under the same environmental conditions (i.e. a single farm) to have a random distribution of Toxoplasma infection. Using breeding records in conjunction with PCR, some families were found to have high Toxoplasma prevalence and abortion while others were free of Toxoplasma infection and abortion (P < 0.01). This supports the notion that Toxoplasma may be transmitted vertically. In humans, we conducted a similar study and showed that Toxoplasma was transmitted from mother to baby in 19.8% of cases. Vertical transmission in Toxoplasma may be more important than previously thought and this knowledge should be considered in any eradication strategies.

Toxoplasma gondii is an obligate intracellular parasite that causes a variety of clinical syndromes, but the infection is more severe in immunocompromised individuals and in cases of congenital toxoplasmosis. This study aimed to verify if the susceptibility to vertical transmission of Toxoplasma gondii is temporally dependent on the preconceptional infection in Calomys callosus. Twelve C. callosus females were infected with 20 cysts of T. gondii ME49 strain and divided into three groups of four animals that were mated after approximately 10 days (group 1), 30 days (group 2), and 50 days (group 3) of infection. The animals were sacrificed from the 17th to 20th day of pregnancy, when placentas and embryos were collected for morphological and immunohistochemical studies, mouse bioassay for evaluating seroconversion and PCR for detecting parasite DNA. Serum samples from C. callosus females and mice used in bioassay were analysed for the detection of IgG antibodies to T. gondii by ELISA. Detection of T. gondii was observed by mouse bioassay and PCR in placentas and embryos from C. callosus females infected around 10 days pre-conception. However, only placentas, but not embryos, from females infected around 30 and 50 days pre-conception showed positivity for parasite DNA and seroconversion by mouse bioassay. In conclusion, this study model shows that vertical transmission of T. gondii may take place when maternal infection occurs within one month before conception, thus demonstrating the time of preconceptional seroconversion that rule out a risk of congenital toxoplasmosis.

To investigate the potential role of endogenous transplacental transmission of Toxoplasma gondii, 31 seropositive ewes presumed to be persistently infected with the parasite and 15 seronegative ewes were mated and monitored throughout pregnancy and lambing. Antibody titres were determined in precolostral sera from the liveborn lambs and in thoracic fluid from the dead lambs. A PCR for the B1 gene of T gondii was applied to the placentas from all the ewes and to the brains of the stillborn lambs. Samples of brain, lung, liver, spleen and heart from the dead lambs were examined by histopathology. No evidence of toxoplasmosis was detected by histopathology or PCR in any of the samples, but low titres of antibody to T gondii were detected in two liveborn, healthy offspring of a seropositive ewe by the immunofluorescent antibody test (3.2 per cent of pregnancies and 4.1 per cent of lambs in the seropositive group). Antibody to specific antigens of T gondii was demonstrated in sera from these two lambs by Western blotting.

Antibodies to Neospora sp. and Toxoplasma gondii were measured in mares and precolostral foals from a farm in Parana State, Brazil. An indirect fluorescent antibody test (IFAT) was used to determine specific antibodies. Three sampling points, 2003, 2004 and at parturition were included in the study, but not all horses are represented at a parturition time point. In 2003, antibodies to Neospora were detected in 17 mares (47%) at 1:50 dilution and in 5 mares (13.8%) at 1:100 dilution. In 2004, antibodies to Neospora were found in 11/36 (30%) horses with titers of 1:50 and in 6 mares (16.6%) at 1:100 dilution. The prevalence of antibodies against T. gondii was 2.7% in mares, either in 2003 and 2004. Evidence for the role of Neospora sp. in equine reproduction failure was not observed in the farm. Immunoglobulin G antibodies to Neospora were found in two of the nine precolostrum foals. Four seronegative foals were born from seronegative mothers, and three seronegative foals were born from seropositive mothers (1:50). Two seropositive precolostrum foals were born from seropositive mothers (1:50). The foals were born clinically normal, and T. gondii antibodies were not detected in their serum samples. The total immunoglobulins values detected on seronegative precolostrum foals were lower than the levels observed in the seropositive foals. This data warrants additional studies to differentiate N. hughesi and N. caninum serologically and determination if these parasites were associated with equine neurological disease and reproductive failure.

It is clear from researching the vertical transmission of Neospora caninum in cattle that the terms 'vertical', 'congenital' and, indeed, 'transplacental' are inadequate for describing two extremely different situations that have fundamentally different immunological, epidemiological and control implications. A similar situation pertains to Toxoplasma gondii in different hosts. We advocate the use of the terms 'endogenous transplacental infection (TPI)' to define foetal infection from a recrudescent maternal infection acquired before pregnancy (and probably prenatally) and 'exogenous TPI' to define foetal infection that occurs as a result of an infection of the dam during pregnancy.

Parasite loads of different tissues were assessed in guinea pig foetus after maternal infection. Twelve female guinea pigs were infected with 100 cysts of the 76 K strain of Toxoplasma gondii by the oral route. Inoculation was performed 20 +/- 5 days (G20) or 40 +/- 5 days (G40) after the beginning of gestation. Gestational age was determined by progesterone assay. Maternal and foetal organ samples were taken 60 days after the beginning of gestation. Parasite loads (from placenta, amniotic fluid (AF), cord blood (CB), foetal brain, liver, lung and spleen) were assessed by a real-time PCR quantification using fluorescence resonance energy transfer (FRET) hybridization probes on the Light Cycler. Congenital transmission was proven by the presence of parasites in blood or tissue samples of the foetus in 84.6% (11/13) and 100% (16/16) of cases after inoculation on G20 and G40, respectively. The quantitative analysis of our results after inoculation at G20 and G40 has allowed us to determinate the positive parasitic loads as a function of the origin of the sample and the period of inoculation. The parasite loads expressed as log (parasite/g) were low in AF and CB samples: 1.49 +/- 0.50 and 1.05 +/- 0.10 at G20 and 1.21 +/- 0.36 and 1.20 +/- 0.42 at G40 respectively. In contrast the placenta and the different foetal tissues had higher parasite burdens: 2.89 +/- 0.54 to 5.30 +/- 0.51 at G20 and 2.81 +/- 0.71 to 3.65 +/- 0.59 at G40. All the placentae were positive for parasites even in the two cases with no proven transmission. Real time quantitative PCR using the hybridization probe was a very sensitive and reproducible technique to study the kinetics of congenital toxoplasmosis in the guinea pig model wich is close to that of humans.

Tachyzoites of Toxoplasma gondii have been found in the milk of sheep, goats, cows and mice and infection by ingestion of raw goat milk has been documented in humans. Lactational transmission from infected cats to their kittens is suspected but the organism has not been detected in the milk. The purpose of this study was to demonstrate the presence of T. gondii in the milk of experimentally infected cats. Pregnant specific pathogen free cats were inoculated orally with T. gondii at various times prior to parturition. Feces were examined for oocyst shedding after sugar solution centrifugation. Milk was collected for polymerase chain reaction (PCR) and bioassay in mice. T. gondii was detected in the milk of five of six cats by either bioassay or PCR.

Captive-bred Mus musculus (house mice) and Apodemus sylvaticus (field mice) were each infected with 50 oocysts of Toxoplasma gondii M1 strain per os and infection in them and their offspring was assessed by polymerase chain reaction (PCR) amplification of the T. gondii B1 gene in brain tissue and by serology, using the modified agglutination test (MAT). The chronically infected female A. sylvaticus (n = 10) and M. musculus (n = 23) were mated at least 6 weeks after infection (and subsequently to produce up to 6 litters) and their pups examined 3 weeks after weaning at 6 weeks of age. By PCR, in offspring of A. sylvaticus and M. musculus respectively, vertical transmission was demonstrated in 82.7% (n = 83) and 85.0% (n = 207) of all pups (N.S., P > 0.05), 95% (n = 21) and 100% (n = 30) of all litters (N.S., P > 0.05), with a mean (+/- S.E.) proportion of each litter infected of 0.87 (0.06) and 0.86 (0.04) (N.S., P > 0.05). There was no change in any of these variables between first and subsequent litters. By serology, whilst MAT suggested 100% vertical transmission in A. sylvaticus, it under-estimated rates of infection in offspring of M. musculus. A limited series of bioassays from M. musculus tissues confirmed the good correlation of PCR and the poor correlation of MAT with mouse inoculation. These results indicate that vertical transmission in A. sylvaticus and M. musculus is extremely efficient and probably endures for the life of the breeding female. This mechanism favours parasite transmission and dispersion by providing a potential reservoir of infection in hosts predated by the cat.

The course of Toxoplasma gondii infection was studied in IL-4-deficient mice from three genetic backgrounds and their wild-type counterparts following peroral inoculation of tissue cysts. Survival rates were significantly reduced in disease-susceptible C57 BL/6 mice and F1 (C57BL/6 x 129Sv) mice deficient in IL-4 compared with wild-type controls. In contrast, this difference was not observed in T. gondii-resistant BALB/c mice. However, brain tissue cyst burdens in IL-4-deficient mice were either equivalent to (C57BL/6 and BALB/c mice) or significantly less (B6/129 mice) than similarly infected wild-type mice. Thus strain-specific differences in the course of T. gondii were demonstrated in the absence of IL-4. The course of T. gondii infection was also compared between B6/129 IL-4-deficient mice and their wild-type counterparts following peroral challenge with 20 tissue cysts on day 12 of pregnancy. Age-matched non-pregnant IL-4-/- and IL-4+/+ mice were also infected to assess the role of IL-4 on T. gondii infection during pregnancy. Disease phenotypes, as measured by mortality, were reversed if infections were initiated during pregnancy compared with non-pregnant infection. Thus significant mortality occurred immediately post partum in IL-4+/+ mothers, while all IL-4-/- mothers survived. Cyst burdens 28 days p.i. were significantly lower in IL-4-/- mothers than IL-4+/+ mothers and both IL-4-/- and IL-4+/+ non-pregnant mice. Congenital disease transmission as measured by foetal death or vertical disease transmission was independent of the presence or absence of IL-4. These studies demonstrate a role for IL-4 in pregnancy-induced immunosuppression and the associated increased susceptibility to T. gondii infection.

To study congenital transmission of Toxoplasma gondii during acute and chronic infections, 4 pregnant Sprague-Dawley rats were each fed 10,000 oocysts of the VEG strain. Toxoplasma gondii was recovered from 33, 55, 83 and 57% of rats (F1) when dams were inoculated at 6, 9, 12 or 15 days of gestation, respectively. Progeny of 15 congenitally infected female rats were examined for T. gondii. Toxoplasma gondii was recovered from tissues of 1 of 155 rats (F2) born to congenitally infected dams. A total of 4 (F2) females were mated; 0 of 40 (F3) rats born to them were infected. None of the acutely infected 4 dams that had given birth to congenitally infected litters produced congenitally infected offspring during the second pregnancy. Thus, unlike mice, evidence for repeated congenital transmission of T. gondii in the rat was found in < 1% of cases. Of the 16 congenitally T. gondii infected pups with demonstrable tissue cysts, 5 were seronegative (< 1:4) in the Sabin-Feldman dye test and 5 were seronegative (< 1:20) in the modified agglutination test by the use of whole formalinized tachyzoites and mercaptoethanol.

Sixteen pregnant queens were inoculated orally with tissue cysts of Toxoplasma gondii, and fetal membranes and offspring were examined for T gondii infection by bioassay in mice. Queens appeared clinically normal, although all shed T gondii oocysts. Toxoplasma gondii was isolated from tissues of 7 of 33 fetuses or kittens from 5 litters (at 13, 23, 26, 27, and 29 postinoculation days) from 8 queens euthanatized between 10 and 31 postinoculation days. Infection with T gondii was found in kittens from all 8 litters from the 8 queens that were allowed to undergo parturition and nurse their kittens. A total of 43 kittens were born to these 8 queens. Toxoplasma gondii was isolated from tissues of 26 of 40 kittens bioassayed; in 3 kittens, tissues were not available for bioassay. Toxoplasmosis was severe in full-term kittens born to 5 queens; all 25 kittens from these litters died or were ill by 24 days of age. Anorexia, lethargy, hypothermia, and sudden death were the most common manifestations. Cytologic examination of peritoneal fluid aspirate samples and determination of hepatic-associated enzyme concentrations in affected kittens, as well as measurement of anti-T gondii antibodies in serum of kittens and queens, were helpful in the diagnosis of neonatal toxoplasmosis. Transplacental transfer of anti-T gondii antibodies was not observed in cats. Toxoplasma gondii oocysts were found in fecal samples of 3 kittens from different litters at 16, 24, and 63 days of age.