Expression of recombinant 35 kDa fragment of VP2 protein of canine parvovirus using Escherichia coli expression system

Research (Published online: 29-06-2021)

35. Expression of recombinant 35 kDa fragment of VP2 protein of canine parvovirus using Escherichia coli expression system

Natnaree Inthong, Sarawan Kaewmongkol, Nattakan Meekhanon, Eukote Suwan, Wanat Sricharern, Khomson Satchasataporn, Rungthiwa Sinsiri, Kaitkanoke Sirinarumitr and Theerapol Sirinarumitr

Veterinary World, 14(6): 1682-1688

ABSTRACT

Background and Aim: Canine parvovirus (CPV) is one of the most common viral infections in dogs, causing acute hemorrhagic gastroenteritis and high mortality. Vaccination effectively prevents CPV infection. However, the currently available CPV vaccines have concerns such as maternal immunity interference, shedding of virus vaccine, and false-positive result based on polymerase chain reaction after vaccination. A subunit vaccine can overcome these problems. This study aimed to express the recombinant 35 kDa fragment of the VP2 protein (consisting of epitopes 1-7) and the recombinant full-length VP2 protein (consisting of epitopes 1-10) and to study the ability of these two recombinant proteins to react with rabbit anti-CPV polyclonal antibodies.

Materials and Methods: The full length and 35 kDa fragment of VP2 gene of CPV were cloned into the pBAD202 Directional TOPOTM expression vector and expressed in E. coli. The recombinant full-length and the recombinant 35 kDa fragment proteins of VP2 were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting.

Results: The recombinant full-length and the recombinant 35 kDa fragment VP2 genes were successfully cloned and expressed. The optimum concentrations of arabinose and induction time for the recombinant full-length and the recombinant 35 kDa fragment VP2 proteins were 0.2% for 6 h and 0.02% for 6 h, respectively. The recombinant full-length and the recombinant 35 kDa fragment VP2 protein molecular weights were approximately 81 and 51 kDa, respectively. The recombinant full-length and the recombinant 35 kDa fragment VP2 proteins specifically interacted with rabbit anti-CPV polyclonal antibodies.

Conclusion: These results suggest that the recombinant 35 kDa fragment and the recombinant full-length VP2 proteins may be useful in developing a CPV diagnostic test or vaccine.

Keywords: canine parvoviruses, Escherichia coli expression system, recombinant protein, VP2 gene.

Trials for preparation and evaluation of a combined inactivated reassorted H5N1 and Escherichia coli O157 vaccine in poultry

Research (Published online: 28-06-2021)

34. Trials for preparation and evaluation of a combined inactivated reassorted H5N1 and Escherichia coli O157 vaccine in poultry

Marwa Fathy El Sayed, Reem A. Soliman, Heba Mohamed Ghanem, Marwa M.S. Khedr, Gina M. Mohamed and Mounir Mohamed Diab El Safty

Veterinary World, 14(6): 1677-1681

ABSTRACT

Background and Aim: Avian influenza (AI), which is one of the major respiratory diseases of poultry, and Escherichia coli (E. coli) have caused major economic losses around the world, including in Egypt. Therefore, in this study, we aimed to produce a vaccine from E. coli O157 and AI H5N1 formulated with Montanide ISA70 for the protection of poultry against both diseases.

Materials and Methods: We divided one hundred 3-week-old chicks into four groups: Group 1 was vaccinated with prepared inactivated AI H5N1formulated with Montanide ISA70, Group 2 was vaccinated with inactivated E. coli formulated with Montanide ISA70, Group 3 was vaccinated with combined E. coli and AI H5N1 formulated with Montanide ISA70, and Group 4 was an unvaccinated control group. We measured the immune response using the HI (hemagglutination inhibition) test, enzyme-linked immunosorbent assay (ELISA), and the challenge test.

Results: We found the three vaccines to be safe and sterile during all periods of examination and observation. The HI test showed that Group 1 exhibited specific antibody titers of 2.3 log2, 4.3 log2, 7.5 log2, 7.8 log2, 8 log2, and 8.1 log2 from week 2 to week 7, respectively, post-vaccination. Group 3 exhibited antibody titers of 3.3 log2, 5.8 log2, 7.8 log2, 8 log2, 8.3 log2, and 8.3 log2 from week 2 to week 7, respectively, post-vaccination. The immune response in both groups reached a high titer at week 6. The combined inactivated E. coli and AI H5N1 vaccine generated a higher immune response than the inactivated AI H5N1 vaccine, and a significant difference exists between the two groups. For Groups 2 and 3, the ELISA antibody titer exhibited its lowest value, 1996.5 and 2036.7, respectively, at week 1 post-vaccination; whereas, both groups exhibited the highest titers, 2227.7 (for Group 2) and 2287.3 (for Group 3), in week 3 post-booster. The ELISA for the combined inactivated E. coli and AI H5N1 vaccine had a higher titer than did the inactivated E. coli vaccine, and a significant difference exists between the two groups. Moreover, the protection rate was higher in Group 3, with 100% for E. coli and 90% for the AI H5N1 vaccine.

Conclusion: Our findings demonstrate that producing a combined vaccine using E. coli and AI H5N1 formulated with Montanide ISA70 is recommended for protection against both diseases.

Keywords: avian influenza (H5N1), Escherichia coli O157, enzyme-linked immunosorbent assay, hemagglutination inhibition test, Montanide ISA70.

Phenotypic characterization and principal component analyses of indigenous chicken breeds in Indonesia

Research (Published online: 28-06-2021)

33. Phenotypic characterization and principal component analyses of indigenous chicken breeds in Indonesia

Dyah Maharani, Fatmawati Mustofa, Aprilianna Putri Z. N. L. Sari, Akhmad Fathoni, Heru Sasongko and Dwi Nur Happy Hariyono

Veterinary World, 14(6): 1665-1676

ABSTRACT

Background and Aim: Understanding the phenotypic characteristics of indigenous livestock breeds is essential for their utilization and conservation. This study aimed to characterize indigenous chicken breeds in Indonesia based on phenotypic traits.

Materials and Methods: Data on eight qualitative and 12 quantitative traits were recorded for 250 chickens from six breeds: Black Kedu, Gaga, Merawang, Nunukan, Pelung, and Sentul. Data were analyzed using descriptive statistics and one-way analysis of variance to test the effect of breed on observed traits. Moreover, principal component analysis (PCA) was conducted separately for each chicken breed. Data on quantitative traits were subjected to Kaiser-Meyer-Olkin, which was computed to test the sampling adequacy and the pattern of correlation among the traits, and Bathlett's tests were used to assess the validity of the factor analysis of each of the datasets and determine whether the partial correlations among traits were small.

Results: We found considerable phenotypic variation in both qualitative and quantitative traits among indigenous chicken breeds. Multicolored plumage (96.40%), wild plumage (39.20%), gold feather flick (51.20%), yellow shank (36.80%), single comb (80.80%), red comb (94.80%), red earlobe (77.60%), and orange eyes (61.60%) were the most common features in the indigenous chickens. In addition, breed had a significant effect on all the quantitative traits that were analyzed (p<0.05). There were higher mean values for all quantitative traits for Pelung chickens than other chickens. In addition, the overall mean values for all quantitative traits in Merawang chicken were intermediate between Pelung chickens and Black Kedu, Gaga, and Nunukan chickens. The PCA showed two principal factors extracted that accounted for 77.80% and 78.38% of the total variance in the original variables for males and females, respectively.

Conclusion: In general, body weight and body measurements, except wattle length, were loaded in PC1 as the primary factors responsible for the variation. The phenotypic variation observed in indigenous chickens in Indonesia could provide valuable basic information for the design of selection and genetic improvement programs.

Keywords: indigenous chickens, phenotypic variation, qualitative traits, quantitative traits.

Assessment of various veterinary drug residues in animal originated food products

Review (Published online: 28-06-2021)

32. Assessment of various veterinary drug residues in animal originated food products

Jagdish Kumar Parmar, Kundan Kumar Chaubey, Vikas Gupta and Manthena Nava Bharath

Veterinary World, 14(6): 1650-1664

ABSTRACT

The veterinary drugs are broad-spectrum antibacterial antibiotics; it uses to cure the animal disease. Many countries have banned veterinary drug residues like nitrofurans metabolites, chloramphenicol. However, the people were administrated veterinary drugs to animals as illegal to increase the milk production in animals for economic benefit. The results of illegally use of veterinary drugs remain as a residue in animal product like milk and it is very harmful to whom consume it cause cancer and allergic for human being which has entered the concern among milk consumers. To control illegal use of veterinary drugs, the government of India has restricted its use in animals. For the identification and confirmation of veterinary drug residues in animal products, analytical techniques such as liquid chromatography and mass spectrometry are available. These are very sophisticated equipments which are available nowadays and their methodologies for the analytical method validation are described by European commission 2002/657/EC. The use of veterinary drugs is a big challenge to effectively identify and authorization of their use. There are so many analytical techniques are using very effectively and taking very less time to protect the consumers from their adverse effects. These techniques take very less time to identify more groups of compounds such as tetracycline, sulfonamides, anthelmintic, and macrolides in single multi-residue method. These methods having validation parameters include system precision, calibration curve, accuracy, limit of detection, and quantification. Therefore, improvement in the existing technologies and accessibility of new screening methodologies will give opportunities for automation that helps in obtaining the results in very less time and improved sensitivity and specificity which contribute to better safety assurance, standard, and quality of various food products of animal origin.

Keywords: antibacterial, antibiotics, food products, methods, techniques, veterinary drug residues.

Investigation of the effects of some processing conditions on the fate of oxytetracycline and tylosin antibiotics in the making of commonly consumed cheeses from the East Mediterranean

Research (Published online: 25-06-2021)

31. Investigation of the effects of some processing conditions on the fate of oxytetracycline and tylosin antibiotics in the making of commonly consumed cheeses from the East Mediterranean

Hussein F. Hassan, Liz Saidy, Rita Haddad, Chadi Hosri, Shady Asmar, Adla Jammoul, Rola Jammoul, Hamad Hassan and Mireille Serhan

Veterinary World, 14(6): 1644-1649

ABSTRACT

Background and Aim: Transfer of antibiotics from raw milk to derived products is directly related to the processes involved in the manufacturing of dairy products, including East Mediterranean cheeses, since these have particular flow diagrams of production. The aim of this study was to assess the effects of skimming, pasteurization, curding, pressing, salting, cheese boiling, and whey acidification/heating on two widely used antibiotics in Lebanon, oxytetracycline (OTC) and tylosin (TYL), in the manufacture of commonly consumed cheeses in the East Mediterranean.

Materials and Methods: Four hundred and fifty kilograms of full-fat bovine milk were spiked with OTC and TYL, then skimmed and pasteurized using holder and high-temperature short-time (HTST) methods. Milk was then processed to make cheeses (23 kg Baladi, 20 kg Akkawi, 20 kg Halloum, and 18 kg Double Cream). Liquid chromatography–mass-spectrometry was used to measure antibiotics. Analysis was performed using Statistical Package for the Social Sciences v25.

Results: Skimming significantly (p=0.015) decreased TYL concentration by 68.6%. OTC degradation during holder (41- 54%) proved to be significant (p=0.015). HTST had a significant (p=0.012) effect on TYL with 32% degradation. Curding step in making Baladi had a significant (p=0.028) effect on OTC only with the concentration increasing by 1.5-fold. Acidification and heating of whey to produce Double Cream decreased significantly (p=0.037) OTC concentration (14.7- 46.3%), while TYL concentration increased significantly (p=0.000) by 300%. Pressing and salting in making Akkawi did not have any significant effect, while cheese boiling in making Halloum significantly decreased both antibiotics.

Conclusion: OTC is transferred to Baladi and Akkawi (curd based) mainly, while double cream (whey based) has a high level of TYL transfer. Hence, people who consume these cheeses excessively could be exposed to high amounts of both antibiotics and thus be prone to their detrimental effect on health.

Keywords: antibiotics, cheeses, liquid chromatography–mass-spectrometry, oxytetracycline, processing, tylosin.

Antioxidant activity and chemical composition of meat from broilers fed diets containing different essential oils

Research (Published online: 25-06-2021)

30. Antioxidant activity and chemical composition of meat from broilers fed diets containing different essential oils

Izabela Lorena Azevedo, Wedson Carlos Lima Nogueira, Anna Christina de Almeida, Lis Lorena Melúcio Guedes, Claudia Regina Vieira, Sergio Henrique Sousa Santos, Carolina Magalhães Caires Carvalho, Francine Souza Alves da Fonseca, Rogério Marcos de Souza and Cintya Neves de Souza

Veterinary World, 14(6): 1638-1643

ABSTRACT

Background and Aim: Broiler meat is susceptible to lipid oxidation due to its high content of unsaturated fatty acids, especially when stored for a long time. Concerning with that problem, we can add natural compounds to the animal feed, as the plant essential oils, which have antioxidant, antimicrobial, and antifungal activities. The objective of this study was to evaluate the antioxidant activity, fatty acid profile, and chemical composition of the meat of broilers fed with diets containing different essential oil, including lemongrass (Cymbopogon flexuosus) and pedestrian tea (Lippia aff. rotundifolia).

Materials and Methods: The experimental design used was a 4×4 factorial scheme (storage time×diets), and each treatment was repeated thrice. The treatments were applied as negative control (without antimicrobial); positive control (ration supplemented with 10 ppm of enramycin and 42 ppm of salinomycin), lemongrass (ration with 120 mg of essential oil of lemongrass for each kilogram of live weight of the animals), and pedestrian tea (ration with 120 mg of essential oil of pedestrian tea for each kilogram of live weight of the animals). Storage was performed after slaughter and evaluated monthly for 4 months.

Results: The inclusion of lemongrass oil and pedestrian tea in the broiler diet did not change the ether extract content of the meat sample obtained from thigh and drumstick. The storage time promotes an increase in the moisture loss of the meat from the 2nd month in all treatments. There was an interaction between the additive and storage time for moisture loss. In the 2nd month of storage, the meat from broilers fed the antibiotic-containing feed had a lower moisture loss than those from broilers in the other treatment groups. The fatty acid profile did not differ statistically between the different treatment groups. The inclusion of additives in the food dropped the lipid oxidation at the 2nd month of storage. From the 3rd month, however, only the essential oils showed antioxidant effect.

Conclusion: Antioxidant activity was positive for treatments that included essential oils. The lemongrass oil and pedestrian tea can be used as antioxidant additives in broiler diets to improve the oxidative stability of thigh-drumstick meat during storage. The results of the study suggest a good prospective for further research with profitability of usage essentials oils examined, for their economical application as feed components in broiler nutrition.

Keywords: broiler, Cymbopogon flexuosus, Lippia aff. rotundifolia, moisture, storage time.

Prevalence of antimicrobial-resistant Streptococcus species among respiratory patients and meat products, and antibacterial effects of oregano oil nanoemulsion

Research (Published online: 24-06-2021)

17. Prevalence of antimicrobial-resistant Streptococcus species among respiratory patients and meat products, and antibacterial effects of oregano oil nanoemulsion

Alshimaa A. Hassanien and Nahed Mahmoud Abdel-Aziz

International Journal of One Health, 7(1): 135-141

ABSTRACT

Background and Aim: Antimicrobial-resistant Streptococcus species causes several diseases in humans with infection recurrence in some cases. Handling of food under unhygienic conditions resulted in transmission of pathogens to food leading to risks for consumers' health. This work studies the possible role of humans in transmission of human pathogens to food products. In this study, we focused on the presence of antimicrobial resistance Streptococcus pyogenes, Streptococcus mitis, and Streptococcus pneumoniae found in respiratory patients and in some meat products. We detected the risk factors for patient's infection and examined the antibacterial effect of oregano oil nanoemulsion (ONE) at different concentrations against the multi-drug resistance strains of streptococci.

Materials and Methods: Microbiological culture and polymerase chain reaction were used to identify S. pyogenes, S. mitis, and S. pneumoniae in 120 throat swab samples and 200 meat products. The disk diffusion method was used for investigating the prevalence of resistance against 10 antimicrobials that are commonly used in the treatment of humans and animals. ONE at four concentrations (0.5%, 1.0%, 2.0%, and 3.0%) with an average particle size of 45.4 nm was identified using transmission electron microscopy. The effect of ONE on Streptococcus species isolates was examined using the well diffusion method. Risk factors for human infection were detected using statistical analysis.

Results: S. pyogenes, S. mitis, and S. pneumoniae were detected in 14 (11.7%), 9 (7.5%), and 7 (5.8%), respectively, of 120 throat swabs of patients. These bacteria were also detected in 11 (5.5%), 8 (4%), and 6 (3%), respectively, of 200 meat products samples. Beef luncheon had the highest infection rate for the three species of streptococci. The majority of these isolates showed resistance to antimicrobials such as tetracycline and amoxicillin/clavulanic acid. ONE had an inhibitory effect on the growth of S. pyogenes, S. mitis, and S. pneumoniae, and its effect was significantly increased at a high concentration (3%), with a mean inhibition zone of 36.45±0.302 mm. Very young and old patients, those with chronic diseases, and those exposed to the infection several times were most susceptible to infection.

Conclusion: ONE may be used as a therapeutic agent at safe and specific doses, and also in the food industry, to prevent the spoilage of food and protect human health.

Keywords: meat products, Oregano oil nanoemulsion, Streptococcus pneumoniae, Streptococcus mitis, Streptococcus pyogenes.