Overall, a very high seroprevalence of toxoplasmosis of 81.4% (95% CI = 77.70, 85.13) was found in women of child-bearing age in Central Ethiopia. This high figure is in agreement with other seroprevalence figures from general or selected populations in different parts of the country that ranged from 60.0 - 97.7%
[14–18, 20]. However, it is much higher than the 20.2% found in pregnant women referred to the National Health and Nutrition Research Center (Addis Ababa) for pregnancy related laboratory examinations
. The high seroprevalence could be attributed to cat density and high rate of oocyst shedding. Recently, Tiao et al. reported that 23.9% (11/46) of the recently infected (IgM positive by ELISA) feral cats in Addis Ababa were also positive for T. gondii oocysts. Tiao et al. also reported 85.4% IgG seroprevalence of T. gondii in 48 feral cats of Addis Ababa. Moreover, the inadequate hygiene, feeding habits and suitable climatic factors for sporulation and survival of oocysts in the environment might have additionally contributed for the high seroprevalence.
Looking into the previous prevalence reports and comparing them with the results of our study, toxoplasmosis in women is still on the rise in Ethiopia perhaps due to the lack of awareness about the disease, as indicated by all respondents in this study (unpublished observations). According to Kapperud et al. the relative importance of the risk factors varies between countries due to differences in cultural patterns and climatic factors affecting oocyst survival. Study area, study population, sample size, age, sensitivities of serological techniques employed, cat densities in the areas and access of cat to contaminate feed and water with oocysts and geographical variability may account for some of the differences in the reported seroprevalence
The Toxoplasma IgG positive and lgM negative results (78.4%, 333/425) suggest past exposure to the parasite or old infection, while positive results for IgM (4.0%, 17/425) are indicators of acute or recent exposures. Though positive IgM results are characteristic markers of recent infections further confirmation to exclude reaction of natural IgM antibody with Toxoplasma antigen is needed
. IgM antibodies can persist for a long time with no risk of congenital infections. Besides, non-pregnant women negative for IgG and IgM antibodies (47/212, 22.2%) are at risk of primary infection and should be monitored for seroconversion in case they become pregnant
. Among the pregnant women of the study only one (0.47%) was IgG negative and IgM positive and 8 (3.8%) were both IgG and IgM positive, which means 4.2% of the women had detectable IgM antibodies during pregnancy with potential risk of congenital toxoplasmosis warranting attention to design preventive measures.
Of the 17 potential risk factors assessed for association with T. gondii seropositivity, study area (P = 0.003), pregnancy status (P = 0.018) and consumption of raw vegetables (P = 0.043) were found to be independent predictors of toxoplasmosis seropositivity.
The association of the seroprevalence with the study areas may be a reflection of the life style of the people that makes them more predisposed to the infection as well as the favorable climatic conditions for T. gondii oocysts to sporulate. The higher seroprevalence of toxoplasmosis in women of Debre-Zeit and Addis Ababa compared to Ambo could probably be attributed to the increasing trend of raw vegetables consumption
 and high usage of water that might have been contaminated with T. gondii oocysts. On the other hand, the higher seroprevalence of toxoplasmosis in Metehara compared to Ambo could be attributed to the combined effect of the presence of cats at home (91.7%), rural background of sampled women (95%), inadequate sanitation and contact with cats (93.3%) rather than raw vegetables consumption (28.3%). Furthermore, the free movement of people of Metehara leading to acquiring of the infection from other areas might additionally explain the situation. The high seroprevalence in women of Debre-zeit (92.6%) could be partly explained by the high seroprevalence reported in sheep (45.3%)
 and goats (40.2%)
 of same area. However, considering the arid and semi-arid nature (unfavorable for oocyst survival) and the relatively low seroprevalence of toxoplasmosis in sheep (13.4%) and goats (15.4%) of Metehara
[32, 33], the high seroprevalence in women of Metehara is controversial. It is possible that women might have higher exposure than animals to cat-originated oocysts since people have a close contact to pet cats. Although consumption of raw sheep and goats’ milk is a common tradition in Metehara pastoralists, only 21.7 % of interviewed women reported such a habit. We recommend further large scale study in Metehara to clarify the high seroprevalence in order to draw reliable conclusions. Variation of T. gondii seroprevalence with geographical location has been noted
[5, 7, 34].
In Ethiopia, antenatal screening of toxoplasmosis is not done unless health professionals have strong suspicion of pregnancy complication in which case patients are referred to the National Health and Nutrition Research Center and other private hospitals and laboratories in Addis Ababa for a better diagnosis. Unlike the reports of Gubre-Xiaber et al.
, who suggested a low risk of T. gondii infection during pregnancy, our findings indicated that a considerable number of pregnant women had recent infections (4.2%, 9/213 IgM positives). The current recent infection rate in pregnant women emphasizes for the need of developing antenatal care programs for toxoplasmosis in Ethiopia. In addition, the IgM and IgG seroprevalence in pregnant women indicates that they are living in a highly contaminated environment. Pregnant women are more susceptible due to immunosuppressant condition of pregnancy where the innate immunity protecting against T. gondii is altered during the 30th and 34th weeks of gestation
. On the other hand, Biedermann et al.
 mentioned that it is difficult to associate the high seroprevalence in pregnant women with immunosuppression as reactivation of latent toxoplasmosis is rare at least in immunocompetent mothers. Although we did not perform CD4+ lymphocyte count of HIV positive pregnant women, there is a chance of reactivation of latent infection with a possibility of congenital transmission. Thus, strengthening the ongoing application of highly active antiretroviral therapy (HAART) and prophylactic treatment at larger scale helps considerably to prevent maternal reactivation and vertical transmission of toxoplasmosis.
Toxoplasma gondii infection was 2.21 times greater in individuals who ate raw vegetables than those who didn’t. The generally poor hygienic method of transport and selling of vegetables coupled with the poor quality water used to wash vegetables might have provided the opportunities for contamination by T. gondii oocysts. Consonant with our results, Njunda et al. from Cameroon, Liu et al.
 from China and Kapperud et al. from Norway reported raw vegetable consumption as an important risk factor for contracting toxoplasmosis. Seroprevalence was higher in women who used unboiled river and well water for drinking purpose (87.3%) than those who used unboiled tap water (75.5%) [P = 0.002], indicating contamination of river and well waters by oocysts from felids’ feces and inadequate water management as reported by Petersen et al.
. These findings are consistent with the already documented scientific knowledge
[2, 37, 39]. Toxoplasmosis was considered as one of water borne diseases
[10, 40, 41].
A relative increase in the seroprevalence was observed with increasing age, as it pertains to the cumulative effect of exposure to the infective stages of the parasite. Once seroconvertion occurred, IgG antibodies persist for life. About 73.3% of the women seroconverted by the time they reach 15 - 20 years of age, most likely following acquisition of oocyst from the environment (as consumption of raw meat for this age group is uncommon). Gubre-Xiaber et al. reported that 75% of Ethiopian children seroconvert before puberty. Several studies have indicated an increase in seroprevalence with age
[3, 10, 13, 20, 34, 39, 42, 43].
The seroprevalence of toxoplasmosis was significantly associated with the presence of domestic cats in the household (84.9%) than in their absence (76.1%) [P < 0.05]. The high seroprevalence observed in households where cats are present suggest a high environmental contamination. Felids are the only definitive hosts responsible for shedding oocysts that contaminate the environment and become infective for a long time in water or soil
. The present finding is in accordance with Acha and Szyfres
 and Negash et al. who reported strong association of seroprevalence and the presence of cats. However, Sroka et al. and Guebre-xabier et al. reported absence of association between seropositivity and presence of cats at home.
Raw meat is popular in Ethiopia and consumption of fresh raw beef, goat or camel meat (typically grass-fed) dipped in a spicy sauce is considered a delicacy. This began centuries ago when Bushmen couldn’t start fires to cook the meat to keep from being seen by enemies
. Eating raw meat (“Kurt” in Amharic language) is sort of a male thing and women just began to eat it more recently. To avoid the problems associated with Taenia saginata, consumption of raw goat meat
 and sometimes mutton is nowadays practiced. Consumption of raw meat depends on culture, habit and economic condition. Despite the deep rooted tradition of raw and undercooked meat consumption and the high seroprevalence of the parasite in sheep and goats
[15, 31, 32, 46–48], no significant association was found between prevalence and raw / undercooked meat consumption (41.6% of all studied women consume raw / undercooked meat) in the current study. On the other hand, earlier studies in Ethiopia
[15–18] and elsewhere
[5, 28] demonstrated significant association between seropositivity and behavior of raw meat consumption.
Although HIV status was not an independent predictor by the final model, HIV positive women (88.4%) are more likely to acquire T. gondii (OR = 2.03, 95% CI: 1.07, 3.85) in the univariate analysis, as compared to HIV negative women (78.9%). This deserves special attention as there is a high chance of reactivation of latent infection and development of toxoplasmic encephalitis
[3, 7].The high seroprevalence of toxoplasmosis in HIV positive women might partly be due to early child hood and teenage infection. In contrast, Biedermann et al.  and Weldemichael et al. reported similar seroprevalence between HIV infected individuals and normal controls.
We are reporting for the first time from Ethiopia that there is a significant association between T. gondii infection and pregnancy, consumption of raw vegetables and use of untreated well and river water.
Committing potential errors by respondents due to recall bias and low health related knowledge (all women have no awareness about health risk of cats to humans) for some of our questions (leading to false negative response), failure to include species of food animals used for meat, failure to make further follow-up and retesting of IgM positive women and selection of sampled women who were volunteers from health institution (that might not necessarily represent the general population) are some of the limitations of our study. Hence, generalization of the results for other geographical areas or entire Ethiopian population needs to be cautiously done.