Seroprevalence of Toxoplasma gondii and associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Pakistan.

Background and Aim: Toxoplasma gondii is an intracellular protozoan that infects humans and animals. This study aimed to estimate the seroprevalence of T. gondii and the associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Mianwali district, Pakistan. Materials and Methods: A total of 350 blood samples were obtained from male and female camels of different ages (≤3 years old, 4-6 years old, and ≥7 years old). To validate T. gondii antibodies, the collected samples were subjected to indirect enzyme-linked immunosorbent assay using purified recombinant micronemal protein 3 as an antibody catching antigen. Results: The prevalence of T. gondii was 50.2% higher in male camels than in female camels (16.5%) (p<0.001). Furthermore, the prevalence of T. gondii in camels was directly proportional to age (p<0.001). It was 63.33% (57/90) in camels of ≥7 years of age, 32.54% in 4-6 years old age group, and 23.08% in ≤3 years old age group. The hematological analysis of infected camels revealed a significant increase in the values of glucocorticoid-remediable aldosteronism, lymphocyte percentage, monocyte percentage (MONO%), corpuscular hemoglobin (MCH), and procalcitonin. Furthermore, substantially higher levels of liver enzymes alanine aminotransferase, aspartate aminotransferase, and the macro-mineral potassium were found in the serum of T. gondii-infected camels. Conclusion: The seropositivity of T. gondii is directly associated with the age and sex of camels, which may be considered as potential risk factors. Furthermore, T. gondii infection directly impacts the hemato-biochemistry of infected camels.

Outbreak investigation and identification of risk factors associated with the occurrence of foot and mouth disease in Punjab, Pakistan.

Foot-and-mouth disease (FMD) is a highly contagious viral disease of domesticated and wild ruminants and pigs that causes enormous economic losses through morbidity, mortality, and trade restrictions. Although the disease is endemic in Pakistan, seasonal outbreaks occur every year throughout the country. This study aimed to investigate FMD outbreaks and to identify the risk factors associated with FMD outbreaks between January and April 2019 in the Punjab province. We performed outbreak investigations (n = 64) for FMD in nine districts of the Punjab province through Divisional Disease Diagnostic Laboratories, Livestock and Dairy Development Department Punjab. Subsequently, we conducted a case-control study including 128 livestock farms (case to control ratio = 1). The data on various plausibly associated factors were collected using a structured questionnaire from each farm. The FMD outbreaks started during the month of January, culminated in February (n = 36, 56.25%) and stopped during the month of April 2019. The most common serotype was O (45.83%), followed by Asia1 (29.17%) and A (13.89%); however, some case farms had mixed infection with serotypes A and O (9.72%) and serotypes O and Asia1 (1.39%). Multivariable analysis revealed that the history of introducing a new animal with unknown FMD vaccination status in the herd (OR =11.51, 95% CI = 3.28 - 40.47), not practicing regular vaccination against FMD (OR = 20.81, 95% CI = 4.59 - 94.35), history of the visit of an animal broker (OR = 9.06, 95% CI = 2.31 - 35.61), distance of the farm to a nearby livestock farm (OR = 6.13, 95% CI = 1.39 - 27.01) and large herd size (OR = 20.79, 95% CI = 2.45 - 176.27) were significantly associated with the occurrence of FMD outbreaks in Punjab province during 2019. In conclusion, improving biosecurity measures, avoiding the introduction of animals without FMD vaccination history and regular vaccination against FMD can significantly reduce the occurrence of the disease on livestock farms in Pakistan.

First molecular evidence of equine granulocytic anaplasmosis in Pakistan.

Anaplasma phagocytophilum (A. phagocytophilum) is an obligate intracellular bacterium that causes equine granulocytic anaplasmosis (EGA) disease in equines. This pathogen has zoonotic potential, which makes it very important to be detected and controlled as early as possible. This study was aimed to assess the molecular prevalence, associated risk factors of EGA along with its effects on various hematological parameters. This study revealed an overall 10.67% prevalence in equine. Horses showed highest prevalence followed by mules and donkeys presenting 11.86, 10.53 and 9.43% prevalence, respectively. The samples were confirmed for anaplasmosis through sequencing. The BLAST queries confirmed very high homology of our isolates with Chinese and Japanese isolates of A. phagocytophilum (Accession no's; KX505303, KY242456 and LC002836). The phylogenetic analysis found the study isolates clustered with each other and this cluster closely resembled Chinese isolate of A. bovis (FJ169957), A. phagocytophilum (HQ872464) and A. phagocytophilum (NR_044762) human isolate from northern Minnesota and Wisconsin. The key risk factors identified for occurrence of EGA in equine species on the basis of univariable analysis were sex of animal, housing type, tick infestation, previous tick history and tick control status, type of acaricides used, rearing system and farm hygiene, respectively. The hematological parameters like Hemoglobin (Hb), Total Leukocyte Count (TLC), Total Erythrocytes Count (TEC), and granulocytes were decreased in diseased animals. The mules showed no typical hematological variations which make sense for its nature as carrier of infection to the susceptible species. This is the first molecular evidence of EGA in Pakistan. The disease needs to be handled seriously as it has zoonotic potential. The animals should be properly attended in disease conditions as leukopenia, neutropenia and lymphopenia can aggravate the condition by making the animal prone to secondary infections.