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Comparative study of productive and reproductive parameters of Holstein Friesian cows in different agroecological zones under subtropical conditions of Pakistan

Masood Ur Rehman1, Syed Muhammad Suhail1, Iqbal Munir2, Rajwali Khan1*

1Department of Livestock Management, Breeding and Genetics, The University of Agriculture, Peshawar, 25130, Khyber Pakhtunkhwa, Pakistan

2Institute of Biotechnology and Genetic Engineering, The University of Agriculture, Peshawar, 25130, Khyber Pakhtunkhwa, Pakistan


The current study aimed to evaluate the impact of three different agroecological zones on the productive and reproductive performance of Holstein-Friesian (HF) cows, along with their comparative analysis of physiological and molecular markers under subtropical conditions of Pakistan. The productive (milk yield and composition) and reproductive (service per conception and calving interval) performance of genetically identical (n=210) HF cows placed across three agroecological zones: irrigated lowlands (Okara), wet (Abbottabad), and dry (Queta) highlands were analyzed. Additionally, heat stress markers (superoxide dismutase, glutathione peroxidase, serum cortisol, bovine heat shock protein 70 (HSP-70), and blood glucose) from five cows in each location (n=15 in total) were also investigated in early and late summers using commercial ELISA and calorimetric kits. Furthermore, the mRNA levels of heat shock protein genes HSPA8 and HSP90AB1 were also quantified in three agroecological zones through qRT-PCR.  The results revealed that cows raised in the wet highlands of Abbottabad exhibited significantly higher (P<0.05) milk production (daily and total lactational yield), and the lowest service period, calving interval and number of services per conception as compared to the cows from the other two agroecological zones. Furthermore, there was a significant effect (P<0.05) of agroecological zones on the heat stress indicators (SOD, GPX, cortisol, blood glucose, and HSP70) profiles and expression of heat shock protein genes in HF cows. The dairy cows from Abbottabad (Wet highlands) showed a significantly lower profile of heat stress indicators as compared to the cows from the other two agroecological zones. It can be inferred that HF cows function better in moderate agroecological zones of subtropical countries.

Keywords: Heat stress indicators, Subtropics, Holstein-Friesian cows, Agroecological zone

Moringa leaf extract enhances the growth and yield characteristics of buckwheat genotypes by modulating the biochemical and physiological activities

Heer Baloch1, Irfan Ali Sabir2, Saadullah Khan Leghari1, Muhammad Sohail Saddiq3*, Pravej Alam4, Shahbaz Khan5*, Esha Mehik Fatima6, Mateen Sajid7, Muhammad Hammad Raza8, Muhammad Arif Hussain1, Muhammad Ayoub1, Rashid Iqbal9

1Department of Botany, Ghazi University, Dera Ghazi Khan, Pakistan

2College of Horticulture, South China Agricultural University, Guangzhou, 510642, China

3Department of Agronomy, Ghazi University, Dera Ghazi Khan, Pakistan

4Department of Biology, College of Sciences and Humanities, Prince Sattam Bin Abdulaziz University, Al- Kharj 11942, Saudi Arabia

5Colorado Water Center, Colorado State University, Fort Collins, CO, USA

6Department of Entomology, Muhammad Nawaz Shareef University of Agriculture, Multan, Pakistan

7Department of Horticulture, Ghazi University, Dera Ghazi Khan, Pakistan

8Department of Agri. Extension and Education, Ghazi University, Dera Ghazi Khan, Pakistan

9Department of Agronomy, Faculty of Agriculture and Environment, The Islamia University of Bahawalpur, Pakistan


Moringa leaf extract (MLE) as a biostimulant has demonstrated success in boosting the productivity of several agronomic crops, but its impact on Buckwheat crops remains unexplored. Buckwheat, recognized as an essential nutritional and functional food crop, often exhibits lower yields compared to major cereal crops grown in similar environments. Therefore, this research aimed to investigate the impact of different concentrations (1%, 2%, and 3%) of MLE on the agricultural performance of common buckwheat (CB) and tartary buckwheat (TB). A pot experiment was carried out according to completely randomized design with factorial arrangements having three replications. Results demonstrated significant improvements in growth parameters (branches, leaves, nodes, and internodes) for MLE-treated plants compared to the control group. Foliar treatment MLE 2% also increased chlorophyll content, improved membrane stability index (MSI) and relative water content (RWC), and enhanced biochemical composition (phenolic compounds, free amino acids, leaf proline, and soluble sugars) in both buckwheat genotypes followed by MLE 3% and MLE 1%. TB produced significantly higher grain yield (0.74 g) as compared to CB (0.43 g). The findings showed that a foliar treatment of MLE 2% led to increased grain yield in both TB (0.97 g) and CB (0.55 g) as compared to control group plants (0.37g TB and 0.22 g CB) respectively. This increase was associated with elevated activities of photosynthetic pigments, phenolic content, RWC, free amino acids, soluble sugars, and catalase in both buckwheat genotypes. In conclusion, MLE application at 2% significantly boosted the agriculture performance of buckwheat, and this study unlocked new insights into optimizing the productivity of the vital food crop.

Keywords: Biostimulant, Free amino acid, Phenolic, Buckwheat, Foliar application, Grain yield

Development of DNA vaccine(s) against Mycobacterium specific genes and prime boost with BCG

Aeman Jilani1, Mirza Imran Shahzad1*, Muhammad Mohsin Zaman1, Areeba Yousaf1, Gildardo Rivera2

1Institute of Biochemistry, Biotechnology and Bioinformatics, The Islamia University of Bahawalpur, Pakistan

2Laboratorio de Biotecnología Farmacéutica, Centro de Biotecnología Genómica, Instituto Politécnico Nacional, México


Tuberculosis (TB), the white plague of Europe is still uncontrolled and fatal in many parts of the world including Pakistan. It is a major cause of morbidity and mortality in human and domestic animals in Pakistan. No new vaccine in the last hundred years has been developed except for a few encouraging results from recombinant and DNA vaccines in the past two decades. Five Mycobacterium specific genes (Rv0379, Rv3914, Rv3006, Rv0432+SP, and Rv0432-SP) were selected to develop DNA vaccine(s). All the constructs were tested on mice using both naked DNA and prime-boost methodologies. Forty-five BALB/c mice were divided into three main groups; DNA vaccine group, BCG Prime boost group, and Control group. Post-vaccine (PV) and post-challenge (PC) immune responses were confirmed through cytokine ELISA and qRT PCR. IFN-γ was additionally checked in plasma as well. Based on cytokine ELISA PC immune responses showed significant differences in TNF-α levels for both naked DNA vaccine groups (Rv0379, Rv3006, and Rv0432-SP) and BCG primed Rv3914 group in comparison to the BCG control group (p<0.05). Based on qRT PCR, IL-6, TNF-α, IFN-γ, and IL-1β showed no significant difference among all the vaccines and BCG control groups (CT range 25- 30). IFN-γ levels in plasma were analyzed PC; two vaccines Rv3006/LppZ and BCG primed Rv0432/SodC-SP (highest mean value 1360.35 pg/ml) have shown significant results (cutoff value 21pg/ml) at 63 days. All the vaccine construct(s) alone or in combination have significant therapeutic effects in comparison to BCG and negative control groups.


Keywords: Mycobacterium tuberculosis, DNA vaccine, Tumor necrosis factor-alpha, Interleukin-6, Interferon-gamma, Interleukin-1beta, Bacille Calmette-Guérin (BCG)

Hemato-biochemical changes, molecular characterization and phylogenetic analysis of the 2022 Lumpy Skin Disease (LSD) outbreak in Cholistan, Pakistan

Tayyaba Asghar1, Mudassar Mohiuddin1*, Ayesha Mohiud Din2,3, Tahira Kamal4, Muhammad Khalid Mansoor1,

Abubakar Siddique5,6,7, Mudasser Habib8, Riaz Hussain9, Muhammad Taslim Ghori10, Hira Hameed1, Humaira Rizwana11,

Islem Abid12, Muhammad Ehsan13, Aroosh Shabbir14

1Department of Microbiology, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan

2Department of Biotechnology, Virtual University of Pakistan, Lahore, Pakistan

3School of Biological Sciences, University of Southampton, Southampton, UK

4National Institute for Genomics and Advanced Biotechnology, National Agricultural Research Centre (NARC), Islamabad, Pakistan

5Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Science and Technology (NUST), Islamabad, Pakistan

6Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, China

7Hainan Institute of Zhejiang University, Sanya, China

8Animal Sciences Division, Nuclear Institute for Agriculture and Biology, Faisalabad, Pakistan

9Department of Pathology, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan

10Department of Clinical Medicine and Surgery, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan

11Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia

12Centre of Excellence in Biotechnology Research, King Saud University, P.O Box 2455, Riyad 11495, Saudi Arabia

13Department of Parasitology, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan

14Institute of Molecular Biology and Biotechnology (IMBB), The University of Lahore, Pakistan


The lumpy skin disease virus (LSDV) is a member of the Capripox genus of the Poxviridae family. It is the causative agent of lumpy skin, a highly contagious disease of cattle, water buffalo, sheep, and goats. In 2022, several outbreaks of LSD were reported in the Cholistan region of Pakistan, which has a large population of livestock living in arid conditions. A total of 230 blood/serum and scab samples were collected from three LSD outbreak locations. Clinically, affected cattle showed acute clinical signs characterized by skin nodules, fever, enlarged lymph nodes, emaciation, and lower leg edema. Hematological findings revealed non-significant changes in red blood cell and white blood cell counts (some animals had leukocytosis while others were leukopenic) whereas, hemoglobin level were significantly low. Platelet count, MPV, PCT, P-LRC, and P-LCC were elevated. Granulocytes were significantly low in LSD affected cattle while lymphocyte counts were significantly high. Serological findings revealed elevated protein levels, along with high creatinine and ALT concentrations. Amplification of DNA-dependent RNA polymerase 30 kDa subunit gene (RPO30) confirmed the presence of LSD virus in all suspected samples. Phylogenetic analysis showed that all Pakistani isolates clustered closely with isolates from neighboring countries. The SNPs differences were less than 20 among these isolates, indicating their close resemblance with each other. It can, therefore, be inferred that our LSD strains might be originated from neighboring Asian countries, that were affected by LSD in previous years.

Keywords: Lumpy skin disease virus (LSDV), Cholistan, Skin lesions, Hematology, Phylogenetic analysis

Influence of Stachys sieboldii Miq. root powder on changes in neural system parameters in growing male rats on a high fat and sucrose diet

Pozdnyakova Yelena1*, Solyanov Dmitry2, Tatina Yelena1, Britko Valeriy1, Omarbekova Nazgul3, Korshukova Marina3

1Department of Biomedicine, Karaganda Medical University, Karaganda, The Republic of Kazakhstan

2Department of Pharmacy, Karaganda Medical University, Karaganda, The Republic of Kazakhstan

3Department of Informatics and Biostatistics, Karaganda Medical University, Karaganda, The Republic of Kazakhstan


The aim of this study is to investigate the effects on the nervous system of the use of Stachys sieboldii root powder, in growing male rats on a high fat and sucrose (HFHS) diet. The animals were divided into three groups: Intact – normal rodent chow, HFHS – high fat and sucrose diet, HFHS + Stachys – high fat and sucrose diet with Stachys supplementation. After 30 days of the experiment, the animals were subjected to the “Open Field” test to study the changes in behavioural responses. Biochemical studies of changes in the concentration of lipid peroxidation products in brain homogenates of growing rats were also carried out. Growing males on the HFHS diet exhibit altered nervous system functioning. Animals show apathy, their motor activity and indicators of exploratory behaviour are reduced and the level of emotional reactivity increases. On the part of biochemical changes – the level of all indicators of lipid peroxidation increases significantly. The introduction of Stachys root powder reduces the negative effect of the diet on the organism of animals – they significantly increase the indicators of exploratory behaviour, increase locomotor activity and reduce emotional reactivity, as well as there is a decrease in the concentration of all products of free-radical oxidation in the brain homogenate. Our study showed that diets high in fat and sucrose had negative effects on growing male rats. The introduction of Stachys sieboldii root powder into the diet improved both behavioural response scores and antioxidant processes in the brain.

Keywords: Stachys sieboldii, Open field, Lipid peroxidation, Conjugated dienes, Ketodienes

Biogenic synthesis and characterization of silver nanoparticles: exploring antioxidant and anti-Inflammatory activities and assessing antimicrobial potential against multidrug-resistant bacteria

Maria Rasool, Muhammad Hidayat Rasool*, Mohsin Khurshid, Bilal Aslam

Institute of Microbiology, Government College University Faisalabad, Faisalabad-38000, Pakistan


Multi-drug resistant (MDR) bacterial infections significantly increase mortality, morbidity, and treatment costs when they persist. Therefore, there is a pressing need to discover, modify, or search for antimicrobial agents with the ability to combat MDR bacteria. Silver nanoparticles used in this study were synthesized by Bacillus subtilis and characterized through different techniques. MDR strains underwent antibacterial activity, antioxidant activity, and time-kill kinetic assays to assess susceptibility to silver nanoparticles. Furthermore, the synergistic impact of silver nanoparticles and antibiotics was examined using the two-dimensional checkerboard method to calculate the Fractional Inhibitory Concentration Index (FICI). Scanning Electron Microscopy (SEM) results revealed a circular shape of synthesized AgNPs, with an average length and area of 76.78 and 47.10 nm, respectively. UV analysis showed an optimum peak at 420 nm. XRD analysis indicated the crystalline nature of nanoparticles with diversity in size. Remarkable antioxidant potential (55% of AgNPs) was observed at a concentration of 1000 µg/ml, while minimum activity (18%) was noted at 62.6 µg/ml. Silver nanoparticles demonstrated a synergistic interaction with the antibiotic Cefixime against Staphylococcus aureus, Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae, with FICI values of 0.37, 0.3, 0.25, and 0.49, respectively, and an additive effect against Pseudomonas aeruginosa, with a FICI value of 0.7. Moreover, this research explores the anti-inflammatory potential of silver nanoparticles (AgNPs) in two distinct models: formaldehyde-induced inflammation and carrageenan-induced inflammation, along with an assessment of their in vitro anti-inflammatory activity. The findings shed light on the multifaceted role of AgNPs in mitigating inflammatory responses, offering promising avenues for therapeutic interventions.

Keywords: Green synthesis, Silver, Bacillus subtilis, Synergistic effect, Cefixime, XRD

Effect of Cordyceps militaris extract on T-lymphocyte, Th2, and Th17 cytokines in vitro and in vivo

Urairat Mongmonsin1, Prapenpuksiri Rungsa2, Withan Teajaroen1, Theerasak Somdee3, Jureerut Daduang4, Sakda Daduang2,5*

1Department of Biomedical Sciences, Faculty of Graduate School, Khon Kaen University, Khon Kaen, Thailand

2Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Khon Kaen University, Khon Kaen, Thailand

3Department of Microbiology, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand

4Department of Clinical Chemistry, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand

5Division of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand


Cordyceps militaris has been used in traditional medicine due to its immune-boosting functions, especially in elderly and cancer patients. The study aims to assess the impact of C. militaris extract on the immune response in LPS-induced splenocytes and male Sprague Dawley rats. The study found that CME contains various bioactive compounds such as adenosine, cordycepin (3′ deoxyadenosine), and phenolic compounds. In vitro studies showed that CME has the potential to enhance CD3+ T cell and CD45RA+ B cell populations in LPS-induced splenocytes with a cell viability of over 80%. Moreover, we found that CME can enhance the immune response in rats by increasing CD3+ T cell proliferation and up-regulating IL-4, IL-6, and IL-17a expression, without affecting the rat’s body weight. The findings suggest that CME could be a promising immune enhancer for patients with weakened immune systems. However, further animal experiments with varying doses are essential to determine the optimal dosage for the successful development of CME as a therapeutic agent.

Keywords: Cordyceps militaris, Immunostimulatory effects, Pro-inflammatory cytokines, Cordycepin

Characterization and identification of bioactive natural products in the ethanol extracts of Acacia nilotica, Melia azedarach, and Euphorbia hirta from Cholistan desert, Pakistan

Sobia Malik1,2, Nuzhat Sial1, Mirza Imran Shahzad3*, Shazia Anjum4, Arshad Javid5, Gildardo Rivera6

1Department of Zoology, The Islamia University, Bahawalpur, Pakistan

2Department of Zoology, Government Sadiq College Women University, Bahawalpur, Pakistan

3Department of Biochemistry, The Islamia University, Bahawalpur, Pakistan

4Department of Chemistry, The Islamia University, Bahawalpur, Pakistan

5Department of Wildlife and Ecology, University of Veterinary and Animal Sciences, Lahore, Pakistan

6Laboratorio de Biotecnología Farmacéutica, Centro de Biotecnología Genómica, Instituto Politécnico Nacional, México



Cholistan desert plants form Fabaceae, Meliaceae and Euphorbiaceae families have always been recognized as an alternate source of medicine and used in different pharmacological activities due to the presence of bioactive secondary metabolites. This study was aimed to characterize bioactive contents in ethanol extracts of Acacia nilotica (whole branch, bark), Melia azedarach (leaves, bark,) and Euphorbia hirta (whole plant). Characterization and composition of secondary metabolites were determined by both chromatographic and non-chromatographic techniques. TLC profile showed maximum spots in Acacia nilotica and M. azedarach. A. nilotica whole branch yielded nine spots for n-Hex, seven spots for DCM, 4 spots for EtAC while A. nilotica bark yielded 4 spots for n-Hex, nine spots for DCM and 4 spots for EtAC. M. azedarach leaves revealed seven spots for n-Hex, six spots for DCM, seven spots for EtAC, while M. azedarach bark revealed seven spots for n-Hex, five spots for DCM, and also five spots for EtAC. E. hirta yielded six spots for n-Hex, two spots for DCM and for EtAC two spots detected. FT-IR spectra showed the characteristic prominent peaks. The maximum number of functional groups were observed in M. azedarach bark, followed by A. nilotica whole branch/bark and M. azedarach whole branch. The least number of functional groups were observed in E. hirta. HPLC analysis was revealed that 9 compounds were majorly quantified in A. nilotica whole branch bark i.e., Gallic Acid, p-hydroxy benzoic acid, Gentisic Acid, Protocatechuic Acid, Catechin, Syringic Acid, Chlorogenic Acid, Vanillic acid, and Epi-catechin. twenty-three compounds were predominantly quantified in M. azedarach leaves bark i.e., Quercetin, Hydroxy ferulic acid hexoside, Rutin, Vanillic Acid, Ferulic Acid, Ferulic acid hexoside II, Feruloylquinic Acid, Myricetin hexoside, Kaempferol -3- O -rutinoside, Kaempferol -3- O – rhamnoside, Procyanidin dimer B, Toosendanin, Quercetin-7-O- glycoside, Kaempferol, Catechin-7-O- glycoside, Apigenin -7-O- glycoside, Kaempferol -7-O-glycoside, Catechin-5-O- glycoside, Capric acid methyl ester, 8- Hexadecene, Phytadiene, γ-n-Amyl butyrolactone, Apigenin, Luteolin, Kaempferol -3-O-glycoside and from E. hirta seven compounds were quantified i.e., Rutin, Gallic Acid, Tannic Acid, Resorcinol, Ellagic Acid, Benzoic Acid, Quercetin. The LCMS scan of A. nilotica whole branch demonstrated the presence of twelve active compounds showing 1.612 -11.183 retention time, fifteen compounds confirmed in A. nilotica bark with rt 0.700 – 14.202, M. azedarach leaves showed only eleven compounds with rt 1.494 -13.031, M. azedarach bark showed 14 compounds in with rt 1.659 – 13.039 and E. hirta revealed eleven compounds with rt 1.557 – 10.884. The GCMS scan of A. nilotica whole branch ten compounds were detected with rt 23.529 – 35.779, A. nilotica bark 9 compounds identified with rt 6.180 – 36.157. Only one compound from M. azedarach leaves identified with rt 34.116, while 5 compounds found in M. azedarach bark with rt 30.740 – 35.379. E. hirta revealed twenty-eight compounds with rt 6.411 – 36.933. The experimental data of studies suggest that the presence of active compound introduce the therapeutic use against infectious diseases and also based on assumption that Cholistan desert medicinal plants are rich source(s) that confers various biological activities.


Keywords: Cholistan desert, Medicinal plants, Ethanol extraction, Liquid-liquid fractionation, TLC, Phytochemical analysis

Differences in gut microbiota and serum Trimethylamine N-oxide (TMAO) levels in patients with colorectal cancer with a small, nested case-control study

Jian Huang1, Xiaohua Chen1, Qinglian Zhong1*

[1] Department of Gastroenterology, The Eighth Affiliated Hospital, Sun Yat-sen University, Shenzhen, Guangdong 518033, China


The dysbiosis of the gut microbiota caused by drug metabolism and diets may influence the gastrointestinal (GI) barrier and their ability for normal attachment and further immunity system, which all could be associated with the medical efficacy of colorectal cancer (CRC) during chemotherapy.

This study aims to investigate Trimethylamine N-oxide (TMAO), a gastrointestinal product readily entering the bloodstream, as a potential risk factor for various diseases, including CRC.

To investigate the relationship between gut microbiota dysbiosis and carcinogenesis in CRC patients, we analyzed taxonomic alterations in the gut microbiota of 77 subjects, including 36 CRC patients and 41 normal controls. We collected samples of the participants’ microbiome from their fecal material and utilized 16S rRNA sequencing to identify the microbial composition. Additionally, to predict the functions of the GI microbiota, Phylogenetic Investigation of Communities by Reconstructing Unobserved States (PICRUSt) was employed. This could serve as a promising biomarker for colorectal cancer. Moreover, the serum level of TMAO between the CRC and healthy controls was also compared, and it was observed that the intestinal microbiome was changed in CRC patients; however, the serum level of TMAO was not correlated with the progression of CRC.

In conclusion, in the present study, instead of relying solely on TMAO, which is a convenient clinical test, we focused on the treatment of CRC by emphasizing the modification of the intestinal microbiome.

Keywords: CRC, Intestinal microbiome, TMAO, 16s rRNA, PICRUSt

Comparative assessment of β-propiolactone, binary ethyleneimine and formaldehyde in inactivating Foot and Mouth Disease virus serotype O

Muhammad Amjad Iqbal1, 2, Noreen Sarwar1*, Sohail Raza1, Sehrish Firyal3, Rabia Riaz4, Afia Muhammad Akram5, Rashad Munir4, Aamir Riaz Khan4, Mobeen Sarwar6, Muhammad Imran Arshad7

1Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore, Pakistan

2Veterinary Research Institute, Lahore, Pakistan

3Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, Pakistan

4Foot and Mouth Disease Research Centre, Lahore, Pakistan

5Department of Zoology, Division of Science and Technology, University of Education, Lahore, Pakistan

6Livestock and Dairy Development, Government of Punjab, Lahore, Pakistan

7Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan


Inactivation of foot-and-mouth disease virus (FMDV) with Formalin (FA) and binary ethyleneimine (BEI) is a slow process and takes long time. The purpose of this study was to evaluate β-Propiolactone (BPL), an alkylating agent that is frequently used in vaccine development and production, as a candidate for the inactivation of FMDV serotype O. Virus was grown on confluent monolayer of BHK-21 cell line. Harvesting was performed between 18 and 20 hours after infection, when CPE was between 90 and 95%. The virus was subjected to inactivation with formalin (0.02%), BEI (2mM, 2.5mM, 3mM) and BPL (0.1%, 0.2 and 0.4%) at 4°C and 37°C. Samples (05 mL/each) were collected after 0, 2, 4, 8, 16, and 24 hours post treatment. The biological titer (TCID50/ml) of each inactivated sample was measured. An innocuity test was used to further confirm the inactivation. RT-PCR was used to detect viral genome damage by amplification of VP1 gene. Data were analyzed by one-way analysis of variance and pair wise comparison was made by post hoc Tukey’s HSD test. Linear regression was used in Microsoft Excel Version 2010. The virus titers after 0.02% formalin treatment were 7.172±0.298 and 4.584±1.362 at 4°C and 37°C respectively. The virus titers were 6.036±0.513, 5.622±0.298, and 5.150±0.449 with 2mM, 2.5mM, 3mM BEI at 4°C and 1.646±1.0210, 1.050±0.644, 0.492±0.492 at 37°C respectively. Inactivation with 0.1%, 0.2% and 0.4% BPL at 4°C resulted in virus titers of 2.386±1.1104, 1.4400±0.9445 and 0.6960±0.6960 respectively. Rapid inactivation of virus with all three BPL concentrations at 37°C gave mean titer of 0.00±0.00. There was a significant difference (P<0.05) among all treatment groups with highest inactivation rates recorded for BPL.

Keywords: FMDV, BHK-21, Inactivation, BEI, BPL, Formalin