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Antibody response of buffalo calves to different levels of Foot and Mouth Disease (FMD) virus immunogen was investigated. Vaccine containing 106.2 units of immunogen/TCID50 of FMD virus (O, A and Asia-1) serotypes induced log2 (1.3± 0.4) units of anti-FMD O Complement Fixing Geometric Mean antibody (FMD O CFT-CGM) titer, log2 (1.4±0.3) units of anti-FMD A CFT-CGM titer and log2 (2.0±0.7) units of anti-FMD Asia-1 CFT-CGM titer. The vaccine containing 2x106.2 units of immunogen of each of the virus serotypes induced log2 (2.2±0.2) units of anti- FMD O CFT-CGM titer, log2 (2.1±0.25) units of anti- FMD A CFT-CGM titer and log2 (3.4±0.8) units of anti-FMD Asia-1 CFT-CGM titer. The vaccine containing 3x106.2 units of TCID50 of each of the virus serotypes induced log2 (5.3 ± 2.0) units of anti-FMD O CFT-CGM titer, log2 (4.6±1.9) units of anti-FMD A CFT-CGM titer and log2 (5.0±2.2) units of anti- FMD Asia-1 CFT-CGM titer. Moreover, buffalo calves (n=3) which were primed and boosted with 60 days interval using vaccine containing 2x106.2 units of immunogen of each of the virus serotype, showed log25.0 and log26.3 units of anti FMD O CFT-GMT antibody titer, log24.6 and log26.0 units of anti FMD A CFT GMT antibody titer, log25.6 and log26.0 units of anti FMD Asia-1 CFT GMT antibody titer, on 30 and 120 days post boosting. Antibody response of buffalo calves was directly proportional to amount of FMD virus immunogen serotypes in the vaccine.

Background: Acute otitis media (AOM) is the second most common disease of childhood. Many factors play role in the causation of AOM. However, the research-based data on risk factors for AOM in infants is still inadequate. Objective were this study aimed to determine the risk factors for AOM in infants with AOM.Methods: This descriptive cross-sectional study was conducted among 189 infant patients with AOM in the otorhinolaryngology outpatient department of allied hospital of Rawalpindi medical university, Rawalpindi. Established inclusion and exclusion criteria and non-probability convenience sampling technique were utilized for the selection of patients. After taking informed consent, data were collected through a self-adapted questionnaire. Descriptive analysis was accomplished through SPSS version 25.Results: AOM was more common among patients who had, an age group with a range of 9-12 months (41.28%), male gender (55.02%), lower birth order (49.73%), mothers with illiterate educational status (53.96%), mothers with the job (57.14%), lower socioeconomic status (59.25%), used bottle-feeding (46.56%), supine position during feeding (61.37%), used a pacifier (67.73%), poor feeding hygiene (58.21%), unvaccinated pneumococcal status (57.15%), and large family (63.50%). In the case of family ear infection and passive smoking, AOM incidence was more frequent among infants whose family had no history of ear infection (69.85%) and no smoking exposure (53.44%).Conclusions: In short, the age group with a range of 9-12 months, male gender, low birth order, illiterate mothers, mothers with the job, lower socioeconomic status, bottle-feeding, supine feeding position, pacifier use, poor feeding hygiene, unvaccinated pneumococcal status, and large family size, all raise the risk of AOM incidence in infants.

Tuberculosis is a devastating disease causing high mortality all over the world, especially in the developing countries.

Mycobacterium tuberculosis (M.tb) causes the current leading infectious disease. Examination of the functional genomics of M.tb and development of drugs and vaccines are hampered by the complicated and time-consuming genetic manipulation techniques for M.tb. Here, we reprogrammed M.tb endogenous type III-A CRISPR-Cas10 system for simple and efficient gene editing, RNA interference and screening via simple delivery of a plasmid harboring a mini-CRISPR array, thereby avoiding the introduction of exogenous proteins and minimizing proteotoxicity. We demonstrated that M.tb genes were efficiently and specifically knocked-in/out by this system, which was confirmed by whole-genome sequencing. This system was further employed for single and simultaneous multiple-gene RNA interference. Moreover, we successfully applied this system for genome-wide CRISPR interference screening to identify the in-vitro and intracellular growth-regulating genes. This system can be extensively used to explore the functional genomics of M.tb and facilitate the development of new anti-Mycobacterial drugs and vaccines.SummaryTuberculosis caused by Mycobacterium tuberculosis (M.tb) is the current leading infectious disease affecting more than ten million people annually. To dissect the functional genomics and understand its virulence, persistence, and antibiotics resistance, a powerful genome editing tool and high-throughput screening methods are desperately wanted. Our study developed an efficient and a robust tool for genome editing and RNA interference in M.tb using its endogenous CRISPR cas10 system. Moreover, the system has been successfully applied for genome-wide CRISPR interference screening. This tool could be employed to explore the functional genomics of M.tb and facilitate the development of anti-M.tb drugs and vaccines.

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a zoonotic pathogen capable of causing severe respiratory disease in humans. Although dromedary camels are considered as a major reservoir host, the MERS-CoV infection dynamics in camels are not fully understood. Through surveillance in Pakistan, nasal (n = 776) and serum (n = 1050) samples were collected from camels between November 2015 and February 2018. Samples were collected from animal markets, free-roaming herds and abattoirs. An in-house ELISA was developed to detect IgG against MERS-CoV. A total of 794 camels were found seropositive for MERS-CoV. Prevalence increased with the age and the highest seroprevalence was recorded in camels aged > 10 years (81.37%) followed by those aged 3.1–10 years (78.65%) and ≤ 3 years (58.19%). Higher prevalence was observed in female (78.13%) as compared to male (70.70%). Of the camel nasal swabs, 22 were found to be positive by RT-qPCR though with high Ct values. Moreover, 2,409 human serum samples were also collected from four provinces of Pakistan during 2016–2017. Among the sampled population, 840 humans were camel herders. Although we found a high rate of MERS-CoV antibody positive dromedaries (75.62%) in Pakistan, no neutralizing antibodies were detected in humans with and without contact to camels.

The objective of this study is to construct a digital gene expression tag profile to identify genes potentially related to immune response in the ostrich. Exposure to boron leads to an immune response in the ostrich, although the underlying mechanism remains obscure. Thus, a dire need of biological resource in the form of transcriptomic data for ostriches arises to key out genes and to gain insights into the function of boron on the immune response of thymus. For this purpose, RNA-Seq analysis was performed using the Illumina technique to investigate differentially expressed genes in ostrich thymuses treated with different boric acid concentrations (0, 80, and 640 mg/L). Compared with the control group, we identified 309 upregulated and 593 downregulated genes in the 80 mg/L treated sample and 228 upregulated and 1816 downregulated genes in 640 mg/L treated sample, respectively. Trend analysis of these differentially expressed genes uncovers three statistically significant trends. Functional annotation analysis of the differentially expressed genes verifies multiple functions associated with immune response. When ostrich thymuses were treated with boron, expression changes were observed in genes predominantly associated with MAPK and calcium signaling pathways. The results of this study provide all-inclusive information on gene expression at the transcriptional level that further enhances our apprehension for the molecular mechanisms of boron on the ostrich immune system. The calcium and MAPK signaling pathways might play a pivotal role in regulating the immune response of boron-treated ostriches.

Objectives: This study was performed to determine the presence of West Nile virus (WNV) in mosquito specimens and human blood donors in Pakistan. Methods: A total of 4150 mosquito specimens were collected using CO2-baited traps from five selected districts of Punjab Province, Pakistan. The mosquitoes were taxonomically identified using standard morphological keys, resulting in 166 pools. In addition, 1070 serum samples were collected from human blood donors. RNA was extracted from mosquito and human samples and screened for WNV using a reverse transcriptase PCR (RT-PCR). Results: None of the mosquito pools tested positive for WNV, whereas three samples from asymptomatic humans tested positive. To determine the WNV strains, partial sequences were compared against a global representation of 23 WNV sequences. The study strains were determined to come from WNV lineage 1. Conclusions: This study is novel in reporting the circulation of lineage 1 WNV in Pakistan. Given its ability to transmit from human to human via blood transfusion, this highlights the urgent need for nationwide surveillance to assess the distribution and impact of WNV in Pakistan. Determining the source of human infection will require more extensive mosquito sampling. Keywords: West Nile virus, Mosquitoes, Human, Pakistan

Rabies, the oldest recorded viral zoonosis in the Indo-Pakistan subcontinent, is a neglected and lingering endemic disease in Pakistan. The review of online available rabies-related journals, papers and reports through platforms and electronic databases have provided the basis for a detailed analysis of the rabies situation in Pakistan. Only published materials related to various research areas of rabies in Pakistan were included and meaningful conclusions were developed to identify major constraints and generate an intellectual discussion on possible solutions. Results revealed 52 studies representing major issues concerning rabies prevention related to topics including, but not limited to: epidemiological investigations (40.38%), prophylactic measures (21.15%), population of wandering dogs (13.46%), public awareness and government interventions (17.30%) and diagnostic surveillance (7.69%). In order to minimize these problems and reduce the prevalence of dog bites or rabies in significant manners, the country direly needs to apply the following actions: a maintained supply of rabies prophylactic measures in public hospitals at subsidized rates, mass dog vaccination at regional levels, enforced responsible animal ownerships, implementing a systematic One Health approach, and diagnostic labs equipped with surveillance mechanisms established in coordination with the livestock and medical departments. This review, which presents up-to-date information on the risk factors and epidemiological features of rabies in Pakistan, provides useful information for scientists, policy makers, and administrative health officials wishing to understand how this deadly disease persists in the absence of effective control measures.

A partir de una sospecha de sarna sarcoptica en tres de cinco conejos albinos europeos (Oryctolagus cuniculus), mantenidos con fines experimentales, se hizo un examen general que revelo areas multifocales de alopecia alrededor de los ojos, las fosas nasales y los labios. Se procesaron cortes de piel y un analisis de PCR con el objeto de identificar los acaros. La histopatologia de los cortes de piel mostro epidermis erupcionada y estrato corneo con una infiltracion de celulas inflamatorias. El raspado o frotis de piel revelo la presencia de acaros tanto en adultos como en huevos. Al examen microscopico, los acaros adultos fueron identificados como Sarcoptes (S.) scabiei (Acari: Sarcoptidae). El analisis de muestras de los tres casos por PCR revelo una banda de 311 pb confirmando la infestacion de S. scabiei en los conejos. La infeccion en conejos con S. scabiei puede representar un problema de salud publica de transmision indirecta debida a la manipulacion de conejos infestados. Hasta la presente, este es el primer reporte de infeccion por S. scabiei en conejos de Pakistan.

This report describes the lower urinary tract disease (LUTD) in four male cats with two different etiologies. All animals were under three years of age and on commercial dry diet. Treatment guidelines prescribed for obstructive and non-obstructive cases were followed. This appears to be the first clinical report on feline LUTD in Pakistan.