Research Article

Detection of Goat and Human Toxoplasmosis Infection using Serological Methods: A Comparative Study

Dunya Abd Al-Malik Mohammed Salih1*, Lina Adil Jebur2, Haitham Abbas Khalaf3, Thaar Mohammed Najim4

1Department of Parasitology, Faculty of Veterinary Medicine, Fallujah University, Iraq; 2Department of Microbiology, Faculty of Medicine, Mustansiriyah University, Iraq; 3Department of Physiology, Faculty of Medicine, Anbar University, Iraq; 4Biotechnology and Environmental Center, Fallujah University, Iraq.

Received | August 21, 2025; Accepted | December 11, 2025; Published | February 11, 2026

*Correspondence | Dunya Abd Al-Malik Mohammed Salih, Department of Parasitology, Faculty of Veterinary Medicine, Fallujah University, Iraq; Email: [email protected]

Citation | Salih DAA-M, Jebur LA, Khalaf HA, Najim TM (2026). Detection of goat and human toxoplasmosis infection using serological methods: A comparative study. J. Anim. Health Prod. 14(1): 358-364.

DOI | https://dx.doi.org/10.17582/journal.jahp/2026/14.1.358.364

ISSN (Online) | 2308-2801

Copyright: 2026 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

INTRODUCTION

In most regions of the world, toxoplasmosis is a widespread zoonotic disease caused by Toxoplasma gondii (T. gondii), an intracellular protozoan parasite that infects humans, animals, including small ruminants (Al-Biatee, 2024; Mustafa et al., 2024), which are an essential part of food and economic security in most countries of the world (Herrero et al., 2013). Sheep and goats contribute significantly to livestock production, making the impact of toxoplasmosis of great veterinary and economic importance due to the reproductive losses it causes (Herrero et al., 2013), such as abortion (Mohammed et al., 2022), fetal abnormalities, and weak births (Fallahi et al., 2018). The transmission of disease to humans through eating undercooked meat or from an infected mother to the fetus during pregnancy (Abdulkhaliq et al., 2017; Stelzer et al., 2019). Together, these mechanisms represent a general health risk, especially to people with weak immunity and pregnant women (Al-Kuraishi et al., 2013; Khalil and Abdulqader, 2019).

The literature in Iraq, including Baghdad, shows a significant prevalence of toxoplasmosis between humans and animals, but these data remain heterogeneous and vary by region (Ali and Alyasiri, 2024). Although there are numerous studies at the population or livestock herd level, there is no updated data available regarding the Abu Ghraib District, which is considered one of the most important areas for goat farming and supporting animal production (Mehanzel, 2012). Also, lacks common estimates linking animal infestation to the handlers who raise them, which limits the effectiveness of targeted control measures (Robertson, 2020). Having such data is essential to enhance understanding of the dynamics of disease transmission. It is also in line with the nature of the region’s diversified production system, which may increase exposure levels (Salman, 2020).

The study is based on biological justifications indicating that age and sexes may affect the probability of exposure to T. gondii as a result of various immune infections and the productive roles of goats and their handlers, from the perspective of one health (Singh et al., 2024), assessing the relationship between animal infections and those responsible for them represents an essential step in understanding the cycle of transmission between Human-animal (Johnson and Johnson, 2021). Hence, integrating these variables provides a more comprehensive analytical framework for understanding infection risks (Huertas-Lopez et al., 2023).

However, there are no contemporary data that jointly estimate the seroprevalence of the disease among goats and their handlers in the Abu Ghraib district, which limits targeted control measures, so, the study aimed that there may be a link between goats and their handlers as a result of participating in the same exposure environment of Abu Ghraib district. The study was designed to detect antibodies, using ELISA testing, and identifying the most prominent risk factors associated with infection.

MATERIALS AND METHODS

Study region and sample population

This study was carried out in the various areas of Abu Ghraib District, located roughly 30-32 square kilometers (km²) or 20 miles west of Baghdad city, Iraq. It’s located at latitude 33.29° north, longitude 44.07° east, and elevation 35-40 meters above mean sea level. Also, it has an area of about 1,232 km² (Al-Jiburi and Al-Basrawi, 2015), as shown in Figure 1 (Mohammad and Awadh, 2023).

 

The population is estimated at 305,000-500,000, includes large rural areas, irrigation canals, and fertile lands that depend on agriculture as the primary source of income, grain and crop production, and animal husbandry under a large-scale agricultural system for meat and milk production (Muhaimeed et al., 2014). Most goats are widely raised on farms, and cases of transmission of parasitic diseases are acquired through constant contact with other animals while grazing and drinking water, mixing of different types of animals, and lack of veterinary services (Madi et al., 2025). Farms containing the goat herds with their handlers were chosen based on the presence of cats, management systems, abortion and diseases associated with the goat herd, etc.

Blood sampling and ELISA examination

One hundred and ninety blood samples were gathered from 100 goats and 90 handlers between July 1st/2024 and May 1st/2025. Goats had been divided into three age groups: ≤ 1 year, 1-2 years, and ≥ 2 years, with a sex of 46 females and 42 males. In addition to 90 handlers’ blood sampled for age groups 18-35 years and 36-50 years, 45 women and 45 men were chosen from the same household in which the goats were located on the farm to ensure the comprehensiveness of the investigation and its coverage of the common human and animal environment. Firstly, approximately 4 ml of blood was drawn by disposable needle syringes from the external jugular vein into vacuum tubes devoid of anticoagulants containing gelatin to collect serum. Each sample is assigned its own data number, represented by sex and age. They were transported in clean plastic containers cooled with ice to the Parasitology Laboratory, College of Veterinary Medicine. It was kept at room temperature to allow coagulation to separate the sera. After centrifugation (3000 rpm/10 min.), then transported in Eppendorf tubes using micropipettes with special data for each and later stored in the freezer at -20 °C until analysis time. After being removed from the freezer to allow it to sit at room temperature. Serum samples were tested for T. gondii-specific IgG antibodies using an ELISA diagnostic kit (IDVET, ID Screen®, Montpellier, France), according to the manufacturer’s protocol. The assay has a reported sensitivity of 100% and specificity of 96% (López-Ureña et al., 2023). Initially, 90 μl of diluted buffer was added to each microwell, and then 10 μl passive control was added to wells A1, B1, C1 and D1. A 10μl sample was added to the remaining well. Then incubated at 45± 4 min/21°C (±5). After that, all wells were emptied and then washed several times with 300μl of washing solution. Conjugate 1x was dilution concentrated conjugate solution of about 1/10 in diluted buffer 3. Another 100μl of conjugate-1X was emptied and washed 3 times with 300µl of wash solution. Then added 100μl of substrate solution to wells and incubated for 15± min./21oC. Also, stop solution (100μl) were added to wells for stopped reaction. All results were read with recording of OD (450 nm) using a plate reader of ELISA. Samples were classified as positive if the optical density (OD) was greater (>) than 50%, doubtful for values between 40-50%, and negative FOR values less (<) then 40%. All percentages were calculated as follows:

Data analysis

All data from the ELISA test results on laboratory investigations were entered and encoded in a spreadsheet file (®). Microsoft Excel 2007 (Aldrich, 2018), we performed statistical analyzes using the statistical version SPSS 21 (IBM Corporation software, Chicago, Illinois, USA). Seroprevalence was calculated by dividing a number of goats and handlers positive for T. gondii antibodies by total number tested. The association of risk factors with seropositivity for T. gondii was assessed using the chi-square (X2) test with corresponding odds ratio (OR) and relative risk (RR). A 95% confidence interval (95%CI) for seroprevalence was calculated to identify statistically significant associations (Andrade, 2023).

RESULTS AND DISCUSSION

Seroprevalence of Toxoplasmosis in goats and handlers

The study results indicated that the overall toxoplasmosis was 55.79% (106/190) by ELISA test. The seroprevalence for 56/100 goats at 59.6% was slightly higher than for 50/90 handlers of 55.56%. Non-significant difference (p>0.05) was observed, the chi-square test χ²=0.0001 are listed in Table 1. As compared with Iran, the infections were found at 6.5% in goats and 14% in humans (Sharbatkhori et al., 2014); in northeastern Algeria, at 94.44% and 53.26%, respectively (Dahmane et al., 2024); and in South Africa, at 11.2% and 8.8%, respectively (Bokaba et al., 2024). The discrepancies observed between handlers and goats in this study may be related to differences in diagnostic tests, occupational exposure levels, and sampling methods when compared with previous studies involving goats and humans. Moreover, there is no consistent pattern indicating that handlers (or meat processors) are always more or less infected than goats across different environments or among professional groups with high exposure to raw meat in slaughterhouse settings (Omonijo et al., 2022).

Toxoplasmosis seroprevalence was highest in goats aged ≥2 years at 65% (26/40), followed by those aged 1–2 years at 54.55% (18/33), and was lowest in goats aged ≤1 year at 44.44% (12/27) (χ² = 2.805; OR = 0.67; RR = 0.815). Female goats showed a higher T. gondii infection rate at 56.86% (30/51) compared with males at 46.94% (26/49) (χ² = 0.339; OR = 1.26; RR = 1.11). However, no significant differences were observed across age groups or between sexes (p > 0.05), as summarized in Table 2.

 

Table 1: Seroprevalence of toxoplasmosis and associated risk factors in goats and handlers.

Sample type	Sample number	Positive (%)	Negative (%)	95% CI	χ²
Goats	100	56 (56)	44(44)	0.464-0.656	0.0001
Handlers	90	50(55.56)	40(44.44)	0.459-0.652
Total	190	106(55.79)	84(44.21)	0.485-0.631
P≥ 0.05; OR=1.018; RR=1.007; df=1

 

Table 2: Seroprevalence of toxoplasmosis in goats by age and sex.

Factors	Positive (%)	Negative (%)	Total (%)	95% CI	χ²
Age
≤1 year	12(44.44)	15 (55.56)	27 (27.0)	0.256-0.632	2.805
1-2year	18(54.55)	15 (45.45)	33 (33.0)	0.366-0.725
≥2 year	26 (65.0)	14 (35.0)	40 (40.0)	0.462-0.838
Total	56 (56.0)	44 (44.0)	100 (199)	0.464-0.656
P-value=0.246 (NS: P≥ 0.05); df=2; OR=0.67; RR=0.815
Sex
Female	30(56.86)	21(41.176)	51(51)	0.454-0.722	0.339
Male	26(46.94)	23(46.938)	49 (49)	0.388-0.674
Total	56 (56)	44 (44)	100 (100)	0.464-0.656
P-value=0.56; (NS: P ≥0.25); OR=1.26; RR=1.11; df=1

 

Compared with previous studies, the age- and sex-related patterns of toxoplasmosis in our study show both similarities and differences. In Baghdad, Iraq, Madi et al. (2025) reported an infection rate of 27.41% in goats aged ≥4 years, compared with 24.13% in those aged ≤1 year. Regarding sex, males showed a notably higher seroprevalence (40.42%) than females (18.88%). In Thi-Qar Province, Iraq, Kareem et al. (2023) recorded a seroprevalence of 56.36% in goats aged ≥2 years and 22.22% in goats aged ≤1 year. Female goats had a higher infection rate (54.54%) than males (35.71%).

In Arabian countries, Chettih et al. (2024) reported a seroprevalence of 40% in goats aged ≥1 year in different regions of Algeria, compared with 36.73% in goats older than 4 years. Infection rates were slightly higher in males (38.89%) than in females (37.84%). In Pakistan, Shah et al. (2013) found that older goats (≥2 years) had a seroprevalence of 54.44%, compared with 20% in younger goats (≤1 year), with females showing higher infection rates (54.5%) than males (26%). Similarly, in Michoacán, Mexico, Alvarado-Esquivel et al. (2013) recorded a higher seroprevalence in older goats aged 49–86 months (22.9%) and in females (16.3%) compared with males (6.3%). The interruption may be due to relationship between T. gondii infection and some biological characteristics of goats, such that both age and sex may play a role in determining susceptibility to toxoplasmosis (Stelzer et al., 2019). An increase in seroprevalence has been observed with age, with adult goats being four times more susceptible to infection than young animals (Ahaduzzaman and Hasan, 2022). Yue et al. (2022) explained that these results are consistent with the observation that older animals are exposed to environmental and disease-causing factors for longer periods, which increases their likelihood of contracting multiple types of infections due to the gradual accumulation of exposure to sources of environmental pollution. Direct contact with infected animals may also play a role in transmitting the oocysts to humans, especially in the absence of personal protective practices (Desta, 2015).

As for the effect of sexes, a higher rate of seropositivity in females is associated with decreased immunity during certain physiological periods, such as periods of stress, pregnancy or the effect of hormones (Shaukat et al., 2024). A strong relationship between occupational exposure and infection registration suggests a similar risk in infection rates and the presence of a common source of infection, whether through soil, feed, or water contaminated with cat faeces, which is the ultimate host of T. gondii (Hatam-Nahavandi et al., 2021).

Higher seroprevalence was exhibited in goats ages 36-50 years at 58.97% (23/39) compared to those 18-35 years at 52.94% (27/51). OR (0.487), RR (0.896), χ² (0.403), and p ≈0.669. Women were largest group at 48.88% (22/45), then men at 42.22% (19/45). OR (1.58), RR (1.27), χ² (0.36), and p-value (0.526), indicates no statistical significance (p>0.05) among ages, sexes, and infection (Table 3).

 

Table 3: Seroprevalence of toxoplasmosis in handlers by age and sex.

Factors	Positive (%)	Negative (%)	Total (%)	95% CI	χ²
Age (Years)
18-35	27 (52.94)	24 (47.06)	51(56.66)	0.36-0.698	0.403
36-50	23 (58.97)	16 (79.48)	39 (43.33)	0.411-0.769
Total	50 (55.56)	40 (44.44)	90 (100)	0.21-1.42
p =0.669 (NS: p≥0.25); OR= 0.487; RR= 0.896; df=1
Gender
Women	22 (48.88)	23(51.11)	45 (50)	0.344-0.634	0.36
Men	19 (42.22)	26 (57.77)	45 (50)	0.28- 0.564
Total	41 (45.55)	49 (54.44)	90 (100)	0.354-0.558
P =0.526 (NS, P≥0.25); OR=1.58; RR=1.27; df=1

 

A comparison between our study and previous reports shows that the presence of T. gondii among handlers in Abu Ghraib, Baghdad, and other professionally exposed individuals is higher than that reported in some countries. For example, in El-Oro, Ecuador, the seroprevalence was 20.16%, with the highest infection rate recorded in individuals aged 24–28 years (25.23%) and the lowest in those over 35 years (16.28%) (Rodríguez et al., 2023). Conversely, in Finland, the prevalence was lower among veterinarians aged <40 years (9.5%) and higher in those aged ≥40 years (19.9%) (Siponen et al., 2019). In Algeria, the prevalence was substantially higher (71.73%), with slight age-related differences: ≤1 year (68.9%), 1–2 years (70.4%), and ≥3 years (74.1%). Female handlers (72.9%) also had a higher seropositivity than males (69.1%) (Dahmane et al., 2020).

These variations in T. gondii seroprevalence among goats and handlers in Iraq and elsewhere may be attributed to multiple risk factors, including differences in animal breed, age, frequency of cat presence on farms, sample size, abortion history, animal susceptibility, and management practices such as hygiene, pasture management, and cat control (Najm et al., 2023; Khan et al., 2024; Dahmane et al., 2024; Rhawy et al., 2024). Additionally, female animals may be more susceptible to infection during pregnancy and lactation due to reduced immunity (Fallahi et al., 2018). Environmental contamination with T. gondii oocysts originating from cat feces deposited on farms, grazing areas, feed, and soil also plays a significant role (Hussain et al., 2017; Mungai, 2021; Stelzer et al., 2019). Large herd sizes, breeding systems, and the close proximity of livestock to cats further increase the risk, and farm operators themselves may be at moderate risk of infection (Bhattarai et al., 2025). Climatic and geographical differences across regions of the world may also contribute to variation in seroprevalence patterns (Ahaduzzaman and Hasan, 2022). T. gondii oocysts reproduce and remain viable for months-years in wet ground, it should not be disputed that domestic and feral cats contribute to environmental pollution, which then exposes humans and animals to infection (Stelzer et al., 2019). Stagnant water basins are an important environmental factor that promotes infection, as water remains in them for long periods without regular change, in addition to relying on a common water source on goat farms, which is a potential causal factor for the transmission of T. gondii infection, as all animals using the same source increase the risk of exposure to infectious eggs (Shapiro et al., 2019; Celi et al., 2022). Evidence suggests that sporozoites may be transmitted through multiple environmental media such as flies, cockroaches, dung beetles, and earthworms, contributing to pollution of the surrounding environment, and can remain stable on the surfaces of fruits and vegetables for long periods (Dixit et al., 2024). Un-sanitized surfaces contaminated with abortion waste inside barns are also important sources of infection, especially in goat farming environments (Mungai, 2021). Infected goats are an important source for meat consumption and are usually slaughtered at a younger age; furthermore, they primarily contribute reservoirs to T. gondii through undercooked meat (Chettih et al., 2024) that appear as their role in foodborne infections, which pose a significant public health risk (Kareem et al., 2023; Khan et al., 2024), or contact with contaminated environments through exposure to the infected goats feces (Rodríguez et al., 2023). The risk of infection for handler lies in consuming contaminated products such as unpasteurized milk or undercooked meat, making the parasite a public health concern, so there is a need to monitor infection in herds periodically, especially with the close relationship between exposure to the parasite and the occurrence of miscarriage (Dahmane et al., 2020), as well as the importance of preventive education to reduce transmission to humans (Al-Biatee, 2024).

Risk factors for toxoplasmosis

Although no statistically significant differences were observed (p ≥ 0.05), the OR (1.018) and RR (1.997) presented in Table 1 still serve as useful indicators for evaluating potential risk factors. In case-control studies, a strong correlation between occupational exposure and infection status is often preferred when assessing disease associations (Ahaduzzaman and Hasan, 2022). This demonstrates that both groups are at risk for infection and that there is a common source of infection, such as soil, feed, or water tainted with cat faeces, which is final host for T. gondii (Hatam-Nahavandi et al., 2021). The statistical difference between goats and handlers (0.001) indicates that goats are more susceptible to infection than handlers, which means that the observed difference due to variability and impact is real, has implications for public health and veterinary practice, and is very useful in peer studies (Schober and Vetter, 2019). The results indicated age-related values in goats [OR (0.67), RR (0.815), and 95% CI (0.464-0.656)], which suggests that older goats (≥2 years) tend to show an increased prevalence of the disease compared to younger goats (≤1 year and 1-2 years); however, this difference was not statistically significant (p ≥ 0.05). In general, it underscores the importance of implementing targeted health interventions within goat populations, with particular attention to younger age groups. In addition, an insignificant trend was observed indicating higher seroprevalence rates in females compared to males, with a probability ratio (1.26), an RR (1.11), and a 95% CI (0.464-0.656), as shown in Table 2. Suggest that the seroprevalence in female goats was higher than in males. According to Table 3, observed higher results among adult handlers compared to young people with OR (0.25), RR (0.896), and 95% CI (0.21-1.42) indicate that adult handlers have more risk factors for seroprevalence than younger people, and women trend toward a higher chance and risk of infection than men. However, the difference is not statistically significant in all ages of handlers.

CONCLUSION

In the Abu Ghraib district, west of Baghdad city, this is the first investigation on Toxoplasmosis infection in goats and their handlers. We find a higher infection rate is influenced by important risk factors including age and sex, which may directly endanger the health and safety of raw meat, dairy products, and animal husbandry and requires more research, such as molecular diagnostic methods that will provide more precise prevalence rate statistics. Programs for food and veterinary health are founded on a thorough knowledge of the epidemiological elements that affect the transmission of illness and aid in efforts to prevent and control disease at the therapeutic and animal levels.

ACKNOWLEDGEMENT

We express our sincere gratitude to the Deanship of Fallujah’s University, Faculty of Veterinary Medicine for their assistance and support with this project.

Novelty Statement

This article provided a careful comparative study between serological detection methods for toxoplasmosis in goats and humans. It is the first of its kind in the region to directly evaluate the effectiveness and accuracy of these methods on both hosts. The results provide accurate indicators of infection, contributing to improved diagnostic and prevention strategies and guiding future studies in parasitology.

AUTHORS CONTRIBUTION

Each author made an equal contribution to the manuscript via blood sample collection, ELISA analysis, data organized, and academic writing.

Funding

There’s no funding.

Ethical approval

All study protocols were approved by the chair of College of Veterinary Medicine, University of Fallujah, Ethics Committee under the Reference number (7) at 25/05/2025.

Generative AI and AI-assisted technology statement

The authors declare that no Genrative AI was used in the creation of this manuscript.

Conflict of interest

The authors have declared no conflict of interests.

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