This study was conducted in four districts (Malakand, Swat, Bajaur and Shangla) of Northern Pakistan to investigate the prevalence, associated risk factors and phylogenetic analyses of Theileria and Anaplasma species in small ruminants. A total of 800 blood samples, 200 from each district, were collected from apparently healthy animals. PCR assays were performed using generic primers for Anaplasma spp. and Theileria spp. as well as species specific primers for A. ovis and T. ovis. Overall infection prevalence was 361/800 (45.1%). Theileria spp. infection prevalence (187/800, 23.3%) was higher than Anaplasma spp. (174/800, 21.7%). Amplified partial 18S rRNA genes were sequenced and enrolled animals were found to be infected by T. ovis (115/800, 14.3%), and at least two more Theileria species (72/800, 9%) were present (T. lestoquardi and T. annulata). All blood samples that were found to be positive for Anaplasma spp. were also positive for A. ovis. Infection prevalence was higher in sheep (227/361, 28.3%) compared to goats (134/361, 16.6%) (p < 0.005). Univariable analysis of risk factors showed that host, age, grazing system and acaricide treatment were significant determinants (p < 0.05) for both Theileria and Anaplasma infections. Multivariable analysis revealed that host, sex, age, tick infestation and grazing system were significant risk factors (p < 0.005) for both pathogens. Phylogenetic analysis revealed variants among the A. ovis and T. annulata samples analysed, indicating that different genotypes are circulating in the field while T. ovis presented the same genotype for the samples analysed.

Ascariasis is a common public health problem of preschool and primary school children in developing countries like Pakistan. The aim of the present study was to determine the prevalence and pattern of Ascaris lumbericoides (A. lumbricoides) infection among children residing in urban areas of Lahore, to provide information on ascariasis to promote awareness and prevention programs between the participants specially on the months or season of higher prevalence. To investigate the prevalence of Ascaris Lumbricoides in the contaminated faecal samples of children residing in urban areas of Lahore, a study was conducted from November 2010 to October 2012 and we collected 3600 stratified faecal samples from six urban study areas. Overall 32/3600 (0.88%) prevalence of fecal samples was found positive for eggs of Ascaris lumbricoides. Area wise highest presence positivity 1.67% was observed in Allama Iqbal Town followed by 1.17% in Samanabad, 1.00% in Wapda Town, 1.00% in Gulberg, 0.50% in Cantt, and the lowest 0.00% in Valencia Town respectively (p < 0.001) The highest month wise positivity prevalence 3/300 (3.33%) (p < 0.001) was observed in the month of September that gradually declined up to 0/300 (0.00%) in the month of March. The results reveal that urban areas of Lahore are susceptible to Ascaris Lubricoides infection and the highest prevalence were observed autumn on the month of September.

Anaplasma marginale is the main etiologic agent of bovine anaplasmosis, and it is extensively distributed worldwide. We have previously reported the first genome sequence of a Mexican strain of A. marginale (Mex-01-001-01). In this work, we report the genomic analysis of one strain from Hidalgo (MEX-14-010-01), one from Morelos (MEX-17-017-01), and two strains from Veracruz (MEX-30-184-02 and MEX-30-193-01). We found that the genome average size is 1.16-1.17 Mbp with a GC content close to 49.80%. The genomic comparison reveals that most of the A. marginale genomes are highly conserved and the phylogeny showed that Mexican strains cluster with Brazilian strains. The genomic information contained in the four draft genomes of A. marginale from Mexico will contribute to understanding the molecular landscape of this pathogen.

The One Health approach looks after animal welfare and demands constant monitoring of the strains that circulate globally to prevent outbreaks. Anaplasma marginale is the etiologic agent of bovine anaplasmosis and is endemic worldwide. This study aimed to analyze, for the first time, the genetic diversity of seven Mexican strains of A. marginale and their relationship with other strains reported. The main features of A. marginale were obtained by characterizing all 24 genomes reported so far. Genetic diversity and phylogeography were analyzed by characterizing the msp1a gene and 5′-UTR microsatellite sequences and constructing a phylogenetic tree with 540 concatenated genes of the core genome. The Mexican strains show 15 different repeat sequences in six MSP1a structures and have phylogeographic relationships with strains from North America, South America, and Asia, which confirms they are highly variable. Based on our results, we encourage the performance of genome sequencing of A. marginale strains to obtain a high assembly level of molecular markers and the performance of extensive phylogeographic analysis. Undoubtedly, genomic surveillance helps build a picture of how a pathogen changes and evolves in geographical regions. However, we cannot discard the study of relationships pathogens establish with ticks and how they have co-evolved to establish themselves as a successful transmission system.

Mycoplasma wenyonii and ‘Candidatus Mycoplasma haemobos’ are bacteria that have been described as significant hemoplasmas that infect cattle worldwide. Currently, three bovine hemoplasma genomes are known. This work aimed to describe the main genomic characteristics and the evolutionary relationships between hemoplasmas, and provide a list of epitopes predicted by immunoinformatics as diagnostic candidates for bovine hemoplasmosis. Thus far, there is no vaccine to prevent this disease that economically impacts cattle production worldwide. Additionally, there is a lack of vaccines against bovine hemoplasmosis. In this work, we performed a genomic characterization of hemoplasmas, including two Mexican strains reported in bovines in the last few years. The generated information is a new scenario about the phylogeny of hemoplasmas. Also, we show genomic features among hemoplasmas that strengthen their characteristic genome plasticity of intracellular lifestyles. Finally, the elucidation of antigenic proteins in Mexican strains represents an opportunity to develop molecular detection methods and diagnoses.