Molecular Epidemiology, Clinical Aspects and Adverse Obstetric Outcomes of Emerging Zoonotic Diseases in Women of Khyber Pakhtunkhwa, Pakistan

Abstract

Introduction: Zoonotic diseases are communicable diseases of vertebrate animals, caused by various pathogens, which can transmit to human through various routes and cause diseases. These diseases are naturally transmitted from vertebrate animals to humans. Among zoonotic diseases, Brucellosis and Toxoplasmosis are zoonotic diseases, which are transmitted from animals and cause various adverse obstetric outcomes in women. Material and Method: The current study was conducted from 2017 to 2020 in Khyber Pakhtunkhwa province of Pakistan. 3586 women of reproductive age (17-45 years) from 7 divisions and 24 districts of KP province of Pakistan were clinically examined and screened for emerging zoonotic diseases (Brucellosis, Toxoplasmosis) using SPAT and RBPT for initial screening of Brucellosis and LAT and ICT assays for Toxoplasmosis, while ELISA and PCR was used as confirmatory assay for Brucellosis and STAT and ELISA for Toxoplasmosis. Hematological parameters like CBC, ESR were determined, Results: Out of 3586 women, 1599 (44.59%) were found positive for zoonotic diseases. Toxoplasmosis was found in 881 (24.56%) and Brucellosis in 718 (20.02%) women. Brucellosis was found in acute, subacute and chronic cases in 428 (59.61%), 239 (33.28%) and 51 (7.11%) women with a significant P<0.05 difference. A significant difference (p<0.05) was found in distribution frequency of zoonotic diseases (Brucellosis, Toxoplasmosis) among various district of the province. Prevalence of zoonotic diseases was significantly P<0.05 high in Karak (72.47%), Chitral (60.95%), Bannu (60.01%) and Kohat (57.81%) district. Assessment of zoonotic diseases was carried out in various seasons in the studied region. A significant difference P<0.05 was found in prevalence of zoonotic diseases in different seasons. Prevalence of zoonotic diseases was significantly P<0.05 high in summer 762 (50.76%) season as compared to spring, autumn and winter season Annual frequency of zoonotic diseases (Brucellosis, Toxoplasmosis) was determined during three year study, but no significant P>0.05 difference was found in prevalence of these diseases (Brucellosis, Toxoplasmosis). Among various socio-demographic characteristics, prevalence of zoonotic diseases (Brucellosis, Toxoplasmosis) was significantly P<0.05 high in (47.75%), (48.51%), (50.41%), (47.21%), (46.61%), (47.91%) and (45.71%) women, who were in age group of 27-36 year, resident of rural area, noneducated, house wives, low socio-economic statues, bad hygienic and who no knowledge about zoonotic diseases. Among various epidemiological factors, a significant P<0.05 association was found in prevalence of diseases with various factors like animals in homes, exposure to animals, handling meat, involvement in processing of dairy products and storing animal dung in homes for agriculture purposes, Prevalence of [x] zoonotic diseases (Brucellosis, Toxoplasmosis) was significantly P<0.05 high in (50.67%), (48.09%), (47.05%), (51.23%) and (50.82%) women, who had animals at homes, exposed to animals, handling meat, involved in processing dairy products and store animals manure in homes, while rate of diseases was low in women, who had no history of these factors. Various laboratory assays like SPAT, RBPT, IgM ELISA and IgG ELISA were evaluated for diagnosis of Brucellosis. SPAT and RBPT significantly P<0.05 detected Brucellosis, while ELISA and PCR significantly P<0.05 confirmed Brucellosis. SPAT was positive in 21.05% cases with sensitivity of 94.29%, specificity 97.28%, PPV 89.67%, NPV 98.55%, while RBPT was positive in 20.21% cases, with a sensitivity of 92.48%, specificity 97.87%, PPV 91.58% and NPV 98.12%. Similarly ELISA was positive in 70.36% cases with sensitivity of 100%, specificity 98.07%, PPV 99.17% and NPV 100%. IgM ELISA was positive in 23.51% cases with sensitivity of 33.15, specificity 98.72, PPV 98.35% and NPV 39.01%. IgG ELISA was positive in 11.66% cases with a sensitivity of 16.44, specificity 99.36%, PPV 98.34%, NPV 33.99%, while ELISA for IgM and IgG was collectively found positive in 35.18% cases with a sensitivity of 50.42, specificity 100%, PPV 100% and NPV 46.63% respectively. Quantitative assessment of SPAT assay was carried out. A highest degree of agglutination 75% was shown by 35.63% women, while lowest agglutination P<25% in 56.9% cases. IgM and IgG antibodies titer was quantitatively estimated by Brucella ELISA assay. A significant difference P<0.05 was found in positivity of IgM and IgG antibodies. IgM was positive in 23.52% women, IgG in 11.66% women, while IgM, IgG were collectively positive in 35.18% women. Brucella specific IgM and IgG were significantly P<0.05 different in acute, sub-acute and chronic cases of Brucellosis Various laboratory assays like LAT, ICT and ELISA were also evaluated for diagnosis of Toxoplasmosis. LAT and ICT assays significantly P<0.05 detected Toxoplasmosis, while no significant difference P>0.05 was found in positivity between these two assays. According to positive cutoff point, LAT assay was found positive in 25.46% cases with a sensitivity of 97.27%, specificity 97.92%, PPV 93.86% and NPV 99.11%. Similarly ICT assay was collectively found positive in 25.51% women with a sensitivity of 96.25%, specificity 97.52%, PPV 92.68% and NPV 98.76%, ICT and ELISA assay was compared for positivity of T. gondii specific IgM and IgG, which was based on qualitative and quantitative detection of these antibodies. A significant P<0.05 difference was found in positivity of IgM and IgG antibodies by these two assays. STAT was used for quantification of T. gondii antibodies. Among 915 women, 3.72%, 68.52% and 27.76% women showed agglutination at 1:32, 1:128 and 1:2048 dilation, which indicate absence of specific antibodies, evolving immunity and recent immunity.