Mycoplasma bovis is a highly contagious pathogen that causes clinical or subclinical mastitis. The present study was aimed for the isolation, molecular characterization and antibiogram determination of M. bovis from raw milk samples. Milk samples were collected randomly from lactating cows and buffaloes from different tehsils of district Faisalabad, Pakistan. Samples were inoculated on modified Hayflick medium and biochemical tests were performed for further confirmation of isolated M. bovis. Out of total 400 milk samples, 184 (46%) samples were found positive for culture method. The 16S-rRNA gene polymerase chain reaction was performed for molecular characterization of isolated M. bovis strains. Out of total 400 milk samples, 240 (60%) positive for M. bovis through PCR method were examined. The 16S-rRNA gene PCR positive isolated M. bovis strains were sequenced and results were compared using Maximum-likelihood method and sequenced strains of M. bovis were aligned and analyzed by Clustal W software. Antibiogram of isolated M. bovis strains was analyzed by disc diffusion assay against eight commonly used antibiotics. Tylosin (30μg) and Tilmicosin (15ug) showed inhibition zones of 32.34 ± 1.10 mm and 17.12 ± 0.93 mm respectively against isolated M. bovis which were found sensitive. Isolated M. bovis was found resistant to other commonly used antibiotics. Statistical analysis revealed that p-value was < 0.05 and the odds ratio was >1.0 at 95% CI. This study complemented the lack of epidemiological knowledge of molecular characterization, comparative effectiveness and resistance trends of isolated M. bovis strains against commonly used antibiotics.

Viral diseases have caused devastating effect on poultry industry leading to significant losses in economy of world. In the presented study, the ability of Newcastle disease virus (NDV), infectious bursal disease virus (IBDV) and avian influenza virus (AIV) to grow in two cell lines was evaluated. Both chicken embryo fibroblast (CEF) and DF-1 cells were used and cytopathic effects (CPE) produced by these viruses were observed. The titer of virus in terms of TCID50 was determined after 24h up to four days for each virus. The same type of CPE was observed for all viruses used in the study in both DF-1 and CEF cells. IBDV showed CPE causing rounding of cells while NDV caused formation of multicellular large nuclei, cell fusion and rounding of cells. Giant cells with inclusions and aggregation of cells with intact monolayer was observed for AIV. In growth kinetic study, higher titer of IBDV and NDV was observed in CEF cells than DF-1 cells while for AIV, DF-1 cells showed higher titer than CEF cells. These results would be useful for furthers comparative studies on growth of different cell lines of various viruses to find a suitability for vaccine production.

Babesiosis is a parasitic disease caused by intraerythrocytic parasites of the genus Babesia, which infect both wild and domestic animals. Merozoite surface antigens (MSAs) have been identified as efficient immunogens in Babesia-infected animals. MSAs play a key role in the invasion process and have been proposed as potential targets for vaccine development. Epitope-based vaccines offer several advantages over whole protein vaccines as the immunogenic proteins are small and can induce both Th1 and Th2 immune responses, which are desirable for protection. However, the MSA, particularly gp45, is polymorphic in Babesia bigemina, posing a challenge to vaccine development. The purpose of this study was to develop a recombinant gpME (gp45-multi-epitope) for a vaccine against Babesia bigemina. B-cell, T-cell, and HLA epitope predictions were used to synthesize the gpME sequence from the consensus sequence of gp45. The gpME sequence was synthesized and cloned in the pET28α vector through the commercial biotechnology company to get pET28-gpME. The plasmid cloned with the gpME sequence comprising 1068 bp was expressed in a bacterial expression system. A band of 39 kDa of rec-gpME was obtained via SDS-PAGE and Western blotting. Rec-gpME @200ng was injected in calves 3 times at 2 weeks interval. The humoral response was evaluated through the indirect ELISA method. The ELISA with rec-gp45 protein showed a significant value of optical density. The recombinant protein containing multiple epitopes from the MSA gp45 may represent a promising candidate for a vaccine against Babesia bigemina.

Azo dye is widely used in the textile industry since it is cost effective and simple to use. However, it becomes a continuous source of environmental pollution due to its carcinogenicity and toxicity. Various methods had been used to remove the azo dye in solution. One of the famous and frequently used is the Fenton process. The Fenton process is one of the advanced oxidation processes where iron catalysed hydrogen peroxide to generate hydroxyl radical. Treating azo dyes in solution requires a catalyst to enhance the process of degradation. Herein, high entropy alloys (HEAs) have been proposed as a catalytic material to enhance the performance of Fenton process for azo dye degradation. HEAs have been reported as a promising catalyst due to its high surface area. The higher the number of active sites, the higher the rate of azo dye degradation as more active sites are available for adsorption of azo dyes. The results have shown that HEAs can be used as a catalyst to fasten the Fenton reaction since the degradation time is proven to be shorter in the presence of HEAs. The method derived from the result of this study will contribute in treating azo dyes for wastewater management in the Fenton process.