First report on molecular surveillance based on duplex detection of Anaplasma marginale and Theileria annulata in dairy cattle from Punjab, Pakistan.

Theileriosis and anaplasmosis are important tick-borne hemoparasites of bovines. The first surveillance study aimed to assess the suitability of duplex PCR for simultaneous detection of Theileria annulata and Anaplasma marginale field infections in Jhang and Rawalpindi districts of Punjab, Pakistan. Cattle blood samples (n = 480) were collected from selected union councils of all tehsils using a multistage sampling technique. The sampling unit consisted of asymptomatic cattle belonging to either age, sex, and breed. Epidemiological data related to host, area, management, and season were collected using a questionnaire. Based on duplex PCR, the overall prevalence of the two concurrent tick-borne pathogens was 19.79% (95/480). Chi-square analysis indicated that age, breed, tick infestation, history of tick-borne diseases, frequency of acaricidial application, and season were significantly associated with tick-borne pathogens. Phylogenetic analysis of A. marginale and T. annulata isolates based on msp1ß and cytochrome b genes, respectively, revealed that nucleotide sequences acquired from these two pathogens are novel, grouped separately from different countries. All our A. marginale isolates showed 88.2 to 80.5% similarity with isolates from Egypt, Israel, Mexico, and lesser homology with South African isolates. Similarly, the phylogenetic tree based on cytochrome b partial sequences of T. annulata revealed that our sequences are closer to those from India and Iran. Based on this first study on concomitant detection of tick-borne pathogens, it can be concluded that mixed infections are endemic in the study districts and mPCR is suitable for detecting concurrent field infections. Simultaneous infections should be considered while performing surveillance and chemotherapeutic trials for better prevention and control of tick-borne diseases.

Seroprevalence of Toxoplasma gondii in one-humped camels (Camelus dromedarius) of Thal and Cholistan deserts, Punjab, Pakistan.

Toxoplasma (T.) gondii is an important zoonotic protozoan infecting humans and a wide range of animals. In this study, we determine the seroprevalence and risk factors associated with the seroprevalence of T. gondii in one-humped camels (Camelus dromedarius) in Pakistan. Camels are still an important mean of transportation in some desert areas in Pakistan. In addition, they are the main source of meat and milk for people in those regions; therefore, they have the potential to transmit T. gondii to humans. In order to estimate the seroprevalence of T. gondii, a total of 897 sera samples were collected from camels in the Thal (n = 359) and Cholistan (n = 440) deserts, along with other districts of Chakwal (n = 44) and Faisalabad (n = 54) Punjab, Pakistan, through convenient and snowball sampling techniques. These samples were then analyzed by an indirect enzyme-linked immune-sorbent assay (ELISA) for the presence of T. gondii-specific antibodies, using purified recombinant micronemal protein 3 (MIC3) as an antibody-catching antigen. Our results showed an overall seroprevalence of T. gondii as 40.1% (Thal = 45%; Cholistan = 35.9%; other districts = 33.7%). Risk factor analysis suggested that infection rate was higher in older animals (70.6%). In addition, female camels carried frequent infection (48.8%) than males (22.4%). What's more, female animals having abortion history showed even higher infection rate (75%) compared to pregnant (68.4%) and non-pregnant (42.4%) animals. Our results reported high seroprevelance of T. gondii in camels in Pakistan which provided important information with respect to public health and disease controls.

Epidemiology, Diagnosis, and Control of Canine Infectious Cyclic Thrombocytopenia and Granulocytic Anaplasmosis: Emerging Diseases of Veterinary and Public Health Significance.

This review highlights the diagnostic methods used, the control strategies adopted, and the global epidemiological status of canine cyclic thrombocytopenia and granulocytic anaplasmosis at the animal-human interface. Canine anaplasmosis is an important worldwide disease, mainly caused by Anaplasma platys and A. phagocytophilum with zoonotic implications. A. platys chiefly infects platelets in canids, while A. phagocytophilum is the most common zoonotic pathogen infecting neutrophils of various vertebrate hosts. Diagnosis is based on the identification of clinical signs, the recognition of intracellular inclusions observed by microscopic observation of stained blood smear, and/or methods detecting antibodies or nucleic acids, although DNA sequencing is usually required to confirm the pathogenic strain. Serological cross-reactivity is the main problem in serodiagnosis. Prevalence varies from area to area depending on tick exposure. Tetracyclines are significant drugs for human and animal anaplasmosis. No universal vaccine is yet available that protects against diverse geographic strains. The control of canine anaplasmosis therefore relies on the detection of vectors/reservoirs, control of tick vectors, and prevention of iatrogenic/mechanical transmission. The control strategies for human anaplasmosis include reducing high-risk tick contact activities (such as gardening and hiking), careful blood transfusion, by passing immunosuppression, recognizing, and control of reservoirs/vectors.