Background: Hard ticks (Ixodidae) are hematophagous ectoparasites that transmit various pathogens to a variety of hosts including humans. Transhumant herds have been involved in the spread of ticks and associated Rickettsia spp., and studies on this neglected topic have been unexplored in many regions including Pakistan. This study aimed to investigate ticks infesting transhumant herds of sheep (Ovis aries) and goats (Capra hircus) in district Shangla, Khyber Pakhtunkhwa, Pakistan. Methods: Of the 144 examined animals, 112 hosts (68 sheep and 44 goats) of transhumant herds were infested by 419 ticks of different life stages including nymphs (105; 25%), males (58; 14%) and females (256; 61%). For molecular analyses, DNA was extracted from 64 collected ticks and subjected to PCR for the amplification of tick 16S rDNA and ITS2 partial sequences and for the amplification of rickettsial gltA and ompA gene sequences. Results: All tick specimens were identified as Ixodes kashmiricus based on morphological features. The obtained 16S rDNA and ITS2 sequences showed 95.7% and 95.3% identity, respectively, with Ixodes kazakstani reported from Kyrgyzstan. In the phylogenetic tree, the sequences clustered with members of the Ixodes ricinus species complex, including I. kazakstani and Ixodes apronophorus. Additionally, rickettsial gltA and ompA partial sequences were 99.7% identical to Rickettsia sp. endosymbiont of Ixodes spp. from Panama and Costa Rica and 99.2% with Rickettsia endosymbiont from the USA. Phylogenetically, the rickettsial gltA and ompA partial sequences from I. kashmiricus clustered with various haplotypes of Rickettsia endosymbiont, which were sister cladded to Rickettsia monacensis. Conclusions: This is the first genetic report of I. kashmiricus and associated Rickettsia sp. Large-scale tick surveillance studies across the country are needed to investigate Ixodes ticks and associated pathogens. Graphical

The online version contains supplementary material available at 10.1038/s41598-026-45336-0.

The mortality rate of women during the gestation period or after delivery is very high worldwide. ToRCH pathogens [Toxoplasma gondii (T.gondii), Rubella Virus (RV), Cytomegalovirus (CMV), and Herpes Simplex Virus (HSV)] are transmitted vertically and are associated with mortality and complications both in the mother and the fetus or children. Therefore, this study is designed to investigate the ToRCH pathogen-induced histopathological changes in placental tissues and associated post-obstetric complications in Pakistani women. Women (N = 68) included in this study were divided into Group A (women with current abortion and bad obstetric history-BOH) and Group B (women with normal delivery and BOH). The venous blood collected was analyzed for ToRCH antibodies through ELISA. A small piece of placenta tissue taken after delivery or abortion was subjected to nucleic acid (NA) detection through PCR using ToRCH pathogen-specific primers. The placental tissue was analyzed for histopathological changes under a microscope. The data obtained were analyzed using percentages and the Chi square test. Overall ToRCH antibody positivity was 64.70% (44/68). Of these, IgM (Immunoglobulin M) was found in 17.65% (12/68), IgG (Immunoglobulin G) in 45.59% (31/68), and one case (1.47%) was seropositive for both IgM and IgG. The NA was detected in 66.18% (45/68) of the placental tissues of the study population. The higher prevalence of antibodies for T.gondii and RV (23.53% each) and NA for RV, CMV, and HSV (17.99% each) was observed in the study population. The ToRCH infection (antibodies and NA) with BOH complications was statistically found to be non-significant (p> 0.05). Overall discrepancies in ToRCH antibodies and NA positivity were found in 86.76% (59/68) of cases, especially for RV (26.47%). Chorionic villitis (CV) in placental tissues was observed in 20.59% (14/68), i.e., 13.23% of Group A and 7.35% of Group B. Of the total, four women with T.gondii, six with RV, and three each with CMV and HSV infection were found with CV. The most common BOH complications observed were recurrent spontaneous abortions (RSAs) (47.05%, 32/68), stillbirth (22.05%, 15/68), congenital abnormalities (14.70%, 10/68), etc. It was observed that RSAs were associated with CV (10.29%) in the study population. The BOH complication and CV were found to be statistically non-significant (p> 0.05). It is concluded that ToRCH pathogens are prevalent in the study population, and CV was the only histopathological change found and associated with BOH complications.