Category Archives: b_original_articles

Original Articles

Enhanced wound healing effects of herbal gel formulations in a rabbit model: a comparative study

Dunia A. Al-Farraj1, Muhammad Kashif2*, Fateh Ullah2, Hafiz Muhammad Ali3, Abdul Qayyum3, Asma Yamin4, Jawaria Aslam5, Shabana Bibi6, Sayed M. Eldin7*, Iftikhar Ali8, Abd El-Zaher M.A. Mustafa1, Mohamed Soliman Elshikh1

1Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia

2Department of Clinical Sciences, University of Veterinary and Animal Sciences, Lahore, Sub-campus Jhang, Pakistan

3Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan

4Department of Zoology, The Govt Sadiq College Women University, Bahawalpur, Pakistan

5Bahawalpur Medical and Dental College, Bahawalpur, Pakistan

6Department of Biosciences, Shifa Tameer-e-Millat University, Islamabad, Pakistan

7Center of Research, Future University in Egypt, New Cairo, Egypt

8Department of Genetics and Development, Columbia University Irving Medical Center, New York, USA

Abstract

The current study was conducted to evaluate the effects of herbal 2% topical gel formulations of either of Allium sativum, Calotropis procera and Prosopis juliflora or their combination compared to an antibiotic cream (Betaderm-N) on healing of full-thickness skin wounds in rabbit. The wound healing (contraction) rate of treated groups was found to be significantly (p<0.05) higher than the positive and negative control groups. The wound treated with A. sativum were healed on 12th day while those treated with P. juliflora or Betaderm-N cream healed on 15th day. The wounds treated with combination gel showed a significantly (p<0.05) higher healing rate and completely healed the wound by 9th day of the experiment and in the histo-pathological examination, there observed an increased number of collagen fibers in dermis of the skin compared to positive and negative controls. Catalase test was used to differentiate S. aureus from other staphylococcal species. S. aureus has golden or creamy colour colonies raised on mannitol salt agar with coagulase positive activity. While the pink colonies raised at Meckonky agar with Indol positive test were of E. coli. By disc diffusion method, the combination of three herbal extracts showed a significantly (p<0.05) higher antibacterial activity against S. aureus and E. coli than other groups and showed a significant increased level of superoxide dismutase (SOD) and reduced glutathione (GPx) at 7th (p<0.05), 14th (p<0.05) and 21st (p<0.01) days of treatments. It was thus concluded that the combined effects of three herbal extracts accelerated the healing process of surgical wound in rabbits due to presence of active metabolites.

 

Keywords: Medicinal plants, Phytochemical analysis, Wound healing, S. aureus, Rabbit

Development of polyclonal antibodies against the recombinant protein of Barley yellow dwarf virus

Muhammad Khalid1, Najam us Sahar Sadaf Zaidi1, Naeem Rashid2, Muhammad Tahir1*

1Atta-ur-Rahman School of Applied Biosciences, National University of Sciences and Technology, Islamabad, Pakistan

2School of Biological Sciences (SBS), University of the Punjab, Lahore, Pakistan

Abstract

Barley yellow dwarf virus (BYDV) particle purification is challenging because of the limited phloem tissue and extremely low viral titers. The current study aimed to generate antibodies against the viral coat protein, without purification of the viral particles. To produce the recombinant coat protein, the genomic region of BYDV encoding the coat protein (CP), was cloned, and expressed in Escherichia coli (E. coli.) BL21 (DE3) strain. Physicochemical characteristics, subcellular localization, and immunogenicity of the BYDV CP (coat protein) were identified using an in-silico approach. The BYDV CP was synthesized by Synbio Technology, USA and cloned into the pET28a (+) expression vector, to produce recombinant fusion coat protein (rFCP-BYDV) in Escherichia coli (E. coli). The recombinant protein produced in inclusion bodies was denatured and purified with on-column refolding by affinity chromatography. Purified protein (rFCPBYDV) was used, as an antigen followed by four weekly intraperitoneal booster doses in mice to develop pAB antisera, which was collected by cardiac puncture, to raise polyclonal antibodies (pAB) in mice. The raised anti-BYDV CP immunoglobulins (IgGs) detected the recombinant BYDV CP even at 100 pg/mL and 1000-time diluted crude extract of proteins from BYDV-infected wheat plant leaves. Results from indirect ELISA titration showed that the anti-BYDVCP antiserum produced in mice had a titer of around 1:10,000. The findings offer a quick and simple immunodiagnostic technique for rapid detection of BYDV. To the best of our knowledge, this is the first report on the production of anti-BYDV CP pAB and their application for the diagnosis of BYDV disease in Pakistan.

 

Keywords: Barley yellow dwarf virus, Wheat, Disease diagnosis, Antibody, ELISA

Metabolic profiles and some physiological traits of three rice cultivars differing in salinity tolerance under salinity stress at the germination stage

Thitiwan Jumpa1, Jutarop Phetcharaburanin2, Manida Suksawat2, Kunlaya Pattanagul3, Wattana Pattanagul1*

1Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen, 40002 Thailand

2Department of Systems Biosciences and Computational Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002 Thailand

3Department of Statistics, Faculty of Science, Khon Kaen University, Khon Kaen, 40002 Thailand

Abstract

This study aimed to investigate salinity’s impact on the germination process and the metabolic profile of germinated seedlings in three rice cultivars known for their varying levels of salinity tolerance. The seeds of three rice cultivars, KDML105 (salt-sensitive), IR29 (salt-sensitive), and Pokkali (salt-tolerant), were germinated in two different treatments: distilled water and a 40 mM NaCl solution. Salinity delayed seed germination and caused root shape abnormalities. The root length of all cultivars was significantly decreased by salinity; however, Pokkali showed the highest root length among these cultivars. Salinity also decreased total soluble sugar, glucose, and fructose content, while starch content was unaffected. Na+/K+ ratio in root and shoot was significantly increased in all cultivars; however, the increase was much lower in the tolerance cultivar. Metabolic profiles revealed several biomarkers of the salt stress response among cultivars and between the control and stressed groups. Gamma-aminobutyric acid was increased in all cultivars. Citric acid, L-lysine, and L-valine were reduced, while uracil was increased in KDML105. L-lysine, succinic acid, and L-methionine were decreased, while adenine was increased in IR29. Oxalacetic acid, L-proline, and urea increased while melatonin decreased in Pokkali.

 

Keywords: Metabolic profiles, Physiological traits, Rice cultivars, Pokkali, Salinity stress

 

Effect of the moisture adsorbents on shallot bulb drying

Muftia Chairin Nissa1, Dewi Qurrota A’yuni1, Setia Budi Sasongko1, Aji Prasetyaningrum1, Mohamad Djaeni1*, Ching Lik Hii2

1Department of Chemical Engineering, Faculty of Engineering, Diponegoro University Jl. Prof H. Soedarto, SH, Tembalang, Semarang, Indonesia

2Food and Pharmaceutical Engineering Research Group, Department of Chemical and Environmental Engineering, Faculty of Engineering, University of Nottingham, Malaysia Campus, Semenyih 43500, Selangor Darul Ehsan, Malaysia

Abstract

Ingredient deterioration, extended drying times, and inefficient energy use are still issues with current shallot bulb drying. As a result, it is suggested that air dehumidification using solid adsorbents increase the driving force in shallot drying. Zeolite and silica, which were used in this study as moisture adsorbents, increased the mass transfer of water from shallot to air. Dehumidification was used to dry about 25 kg of fresh shallots for 4 hours at temperatures of 30 °C, 40 °C, and 50 °C, with an average air velocity of 7.8 m/s. Results indicated that using adsorbents throughout the drying process could speed up the reduction of moisture content. In addition, Page’s model predicted accurately the rate of shallot bulb drying for any variable. The total phenolic compounds (TPC) decreased at higher drying temperature and longer drying time. The addition of zeolite can keep the TPC high. Meanwhile, the thermal energy efficiency rose at higher temperatures. Response surface methodology (RSM) determined that air dehumidified by zeolite at a drying temperature of 50 °C produced the best of shallot drying results.

Keywords: Shallot bulbs, Adsorbent drying, Total phenolic, Mathematical modeling

Transcriptome analysis of flower development and mining of genes related to the flower development in Oncidium

Nengyan Fang, Yuanhua Luo, Ronghui Fan, Xiuxian Ye, Minling Huang*, Huaiqin Zhong*

Institute of Crop Sciences, Fujian Academy of Agricultural Sciences (FAAS), Fujian Engineering Research Center of Characteristic Floriculture, Fuzhou, 350013 China

Abstract

Oncidium, a kind of orchid plants characterized with unique flower, is one of the four tropical orchids with high ornamental value and favored by consumers. However, our understanding about the molecular basis of its flower development is still limited. Here, we collected Oncidium tissues at different developmental stages for RNA-seq. A total of more than 621 million clean reads were generated. 134,640 unigenes were assembled, and 54,221 unigenes were annotated. The number of DEGs (differentially expressed genes) was the largest in the comparison group F2-vs-F3 (F2 stage compared with F3 stage in flowers). The GO (Gene Ontology) terms and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways enriched for the specific DEGs were diverse at different stages. The common pathways enriched in multiple comparisons were also obtained. 11 key genes were obtained in cutin, suberin and wax biosynthesis pathway. Moreover, 32 candidate genes related to flower development were screened. Most of them presented tissue-specific expressions, especially MADS-box genes described by the ABCDE model. In all, the present data provides a valuable resource for dissecting the molecular mechanism of Oncidium in regulating flower development.

 Keywords: Candidate genes, DEGs, Flower development, MADS-box genes, Oncidium, RNA-seq

Sorting effects of X/Y sperm on in vitro fertilization of Belgian blue bulls

Peng Niu1, Fei Huang1, Xin Liu2, Bo Liu2, Jie Wang1, Di Fang1, Ahrar Khan4,5*, Qinghua Gao1,2,3*

1College of Life Sciences and Technology, Tarim University, Alar, Xinjiang 843300, China

2College of Animal Science and Technology, Tarim University, Alar, Xinjiang 843300, China

3Key Laboratory of Tarim Animal Husbandry Science and Technology, Xinjiang Production & Construction Corps, Alar, Xinjiang 843300, China

4Shandong Vocational Animal Science and Veterinary College, Weifang, China

5Faculty of Veterinary Science, University of Agriculture, Faisalabad-38040, Pakistan

Abstract

There is a large market in China for the production of Belgian blue bulls, which would benefit from semen flow sorting for animal sex control. The sperms of the Belgian blue bulls were separated by flow cytometry, and then the quality of sorted sperm was tested by in vitro fertilization (IVF) and artificial insemination (AI). The fresh semen of 8 Belgian blue bulls was individually collected and processed for sorting. Sperm sorting was carried out using MOFLO cell sorter and cryopreservation. There were no significant differences in sperm motility, acrosome integrity, and DNA integrity between sex-sorted and non-sorted sperm (p > 0.05). The purified sperm with higher viability was fertilized with mature oocytes in vitro, co-cultured to cleavage and blastocyst stage, embryo sex was identified by nested PCR amplification of AMEL gene, and there was no significant difference between sorted sperm and non-sorted sperm (p > 0.05). For artificial insemination, the pregnancy rate using non-sorted sperm was higher than for sorted sperm (p < 0.05). After delivery, the sex ratio of offspring using X- and Y-sperm was 100% and 90.9%, respectively, with a significant deviation from conventional semen (p < 0.05). The birth weight of male and female calves in the control group was similar to that in the sex-sorted sperm (p > 0.05). In summary, after artificial insemination using sex-sorted sperm, normal Belgian blue calves with the predicted gender can be produced. It is of great significance to improve the commercial promotion and production efficiency of semen after sorting Belgian blue bulls.

Keywords: Belgian blue bull, Flow cytometry, In vitro fertilization, Artificial insemination, Sex ratio

Immune enhancing activity of garlic polysaccharide, selenizing Codonopsis pilosula compounds in chickens

Li Liang-liang1,2, Liu Kuan-hui3, Xie Shi-hui3, Li Ting3, Zhu Guang-huang3, Mikhlid H. Almutairi4, Muhammad Zahid5, Mushtaq Ahmad Gondal6, Qudratullah6, Khalid Mehmood5*, Yi Wu1*

1College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China

2College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China

3Wuhu Institute of Technology, Wuhu, China

4Zoology Department, College of Science, King Saud University, Riyadh, Saudi Arabia

5Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan

6Cholistan University of Veterinary and Animal Sciences Bahawalpur, Pakistan

Abstract

Codonopsis pilosula polysaccharide is a main ingredient of Codonopsis. The key component in Condoposis is identified as Codonopsis pilosula. This Codonopsis has multiple functions including stimulation of spleen, lungs and tonifying the middle Jiao and Qi. Allium sativum has functions of regulating vital energy, appetizing, detoxification, and disinfection. Polysaccharide of the bulbous is key component to perform the functions of this bulbous. In current study, the immune enhancing activity of GP, sCPP and GP-sCPP through in-vivo and in-vitro trials was evaluated. For in-vivo trials, a total of 150 one day old white roman chicks were randomly divided into five groups, namely immune control (VC), blank control (BC), GP-sCPP, sCPP and GP groups. Chicks in group GP-sCPP, sCPP and GP was intramuscularly treated with 0.5 mg, 0.5 mg and 4.5 mg of polysaccharide, respectively, in all experimental days, and chicks in VC and BC were treated with equal volume of normal saline injections daily. All chicks were bred according to the standards for the rearing of laying hens. For in-vitro trials, lymphocyte cells were cultured in a 12-well plate along with different concentrations of GP-sCPP, sCPP and GP were supplemented in the cells and incubated at 37.5℃. After that, MTT was employed to measure the cytotoxicity of those polysaccharides. The results showed that GP, sCPP and GP-sCPP could increase immune-enhancing activities by releasing cytokines of IL-2 and IFN-γ. In-vivo experiments in chicks showed significantly higher serum antibody titer, T lymphocyte proliferation, and IFN-γ, IL-2 and IL-6 levels in GP-sCPP group. These results demonstrated that GP-sCPP can significantly promote immune enhancing activity, which could be recognized as a novel candidate for new type of immune modulator.                          

Keywords: Lymphocyte proliferation, Codonopsis Pilosula, Chickens, mRNA expression, Antibody titer, Cytokines

Sciadopitysin attenuates paraquat induced renal toxicity by modulating Nrf-2/Keap-1 pathway in male albino rats

Muhammad Umar Ijaz1*, Ayesha Kalsoom1, Ali Hamza1, Nazia Ehsan1

1Department of Zoology, Wildlife and Fisheries, University of Agriculture, Faisalabad, Pakistan

Abstract

Paraquat (PQ) is a herbicide that has the potential to instigate nephrotoxicity in animals and human. Sciadopitysin (SPS) is a biflavonoid that is extracted from Taxus cuspidate and displays diverse biological activities including anti-oxidant, anti-inflammatory and anti-apoptotic. Therefore, the present investigation was designed to evaluate the mitigative potential of SPS against PQ prompted renal toxicity in albino rats. 48 male albino rats were divided into 4 groups, such as control group, PQ treated group (5 mgkg-1), PQ + SPS co-treated group (5 mgkg-1 and 2 mgkg-1 respectively) and only SPS treated group (2 mgkg-1). The exposure of PQ significantly reduced Nrf-2 as well as anti-oxidant enzymes expression, while increasing Keap-1 expression. Moreover, anti-oxidant enzymes such as, glutathione reductase (GSR), superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione-S-transferase (GST), heme oxygenase-1 (HO-1) and glutathione (GSH) activities were decreased. However, in PQ-treated rats malondialdehyde (MDA) and reactive oxygen species (ROS) contents were significantly increased. PQ exposure also increased the serum level of urea, urinary protein, urobilinogen and creatinine while decreased creatinine clearance and albumin protein levels. Moreover, KIM-1 and NGAL levels were also increased in PQ exposed rats. Additionally, inflammatory indices including nuclear factor kappa-B (NF-κB), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) interleukin-1β (IL-1β) and cyclooxygenase-2 (COX-2) activity were increased in PQ administrated rats. Besides, it escalated the Bax and Caspase-3 expression. Contrarily, a substantial decrease was observed in anti-apoptotic marker, Bcl-2 expression. The exposure of PQ also induced significant histopathological damages in renal tissues. Nevertheless, SPS supplementation recovered all these damages due to its anti-apoptotic, anti-oxidant and anti-inflammatory nature.

Keywords: Paraquat, Sciadopitysin, Renal damage, Oxidative stress, Inflammation

The effects of Cryptosporidium infection on gut fungi and enzyme abundance in Sus domesticus

Wang Lei1,3, Lu Sijia1, Zou Wen1, Sun Na2, Muhammad Usman Saleem5, Mudassar Nazar6, Mohammad Farooque Hassan7, Farid Shokry Ataya8, Kong Qinghui4, Li Kun1*

1Institute of Traditional Chinese Veterinary Medicine & MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China

2Shanxi key lab. for modernization of TCVM, College of Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, Shanxi, China

3College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China

4Key Laboratory of Clinical Veterinary Medicine in Tibet, Tibet Agriculture and Animal Husbandry College, Linzhi, Tibet, China

5Department of Biosciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan

6University of Agriculture Faisalabad, Sub-Campus Burewala 61010, Pakistan

7Shaheed Benazir Bhutto University of Veterinary & Animal Sciences, Sakrand, Sindh, Pakistan

8Department of Biochemistry, College of Science, King Saud University, Riyadh 11451, Saudi Arabia

Abstract

Pigs are known to be natural hosts of Cryptosporidiosis, which poses a serious threat to public health. According to the World Health Organization (WHO), Cryptosporidiosis is the major cause of severe diarrhea which causing death in infants. Many researchers suggest that gut fungi have an indispensable function in host metabolism and immunity. However, few studies have been performed to understand how Cryptosporidium infection induces alterations in the intestinal fungal communities of Tibetan pigs. Therefore, fecal samples from Cryptosporidium infected and Healthy Tibetan pigs were examined by internal transcribed spacing (ITS) gene amplification sequencing. Results showed that a total of 2 phyla, 5 classes, 9 orders, 10 family, 13 genera of fungi were detected from both the H (Healthy Tibetan pigs) and INF (Cryptosporidium infection pigs) groups. The results manifested that proportion and profile of the total fungal population obviously changed under the Cryptosporidium infection, marked by a reduction in the abundance of beneficial fungi, i.e., Leotiomycetes (p < 0.05), Aspergillaceae (p < 0.05), Penicillium (p < 0.05), Xenoacremonium (p < 0.05) and Eurotiales (p < 0.05) and an increase in the abundance of disease-causing fungi that threaten health, i.e. Colletotrichum (p < 0.05) and Clarireedia (p < 0.05). In addition to these changes, some enzymes including Arabinose-5-phosphate isomerase, Quercetin 3-0-methyltransferase and Amylosucrase were found to be significantly altered (p < 0.01) after Cryptosporidium infection using fungal function prediction analysis. This study also focused on the Cryptosporidium infection in Tibetan pigs in terms of gut fungal diversity, composition, and the abundance of enzyme. It is providing a better understanding of Cryptosporidium infection in Tibetan pigs and insights for further development of therapeutics against Cryptosporidium from the gut fungal perspective.

Keywords: Sus domesticus, Tibetan pig, Cryptosporidium, Gut fungi, Enzyme, Microbiota

Biological activity of leaves of three Morinda species detected by stimulation and suppression of gene expression of collagen, elastin, melanin and other related genes

Runglawan Sudmoon1, Sanit Kaewdaungdee2, Tawatchai Tanee3, Pornnarong Siripiyasing4, Arnold Ang5, LiTing Lee5, Xue Jing Wong5, Shiou Yih Lee5, Arunrat Chaveerach2*

1Faculty of Law, Khon Kaen University, Thailand

2Department of Biology, Faculty of Science, Khon Kaen University, Thailand

3Faculty of Environment and Resource Studies, Mahasarakham University, Thailand

4Faculty of Science and Technology, Rajabhat Mahasarakham University, Thailand

5Faculty of Health and Life Sciences, INTI International University, Malaysia

Abstract

Morinda coreia, M. elliptica and M. tomentosa leaves were investigated. α-EG and pinoresinol were found by GC-MS and GC-FID. MTT and comet assays did not reveal toxicity in normal PBMCs and DHFa cells, but the ethanol and hexane M. coreia and M. tomentosa extracts were toxic to CHL-1 cancer cells. Biological activities were detected by gene expression through qRT-PCR. Ethanol M. tomentosa extract can stimulate collagen type I, II, III and elastin creation through expression of COL1A, COL1A2, COL2A1, COL-II, COLL2, COL3A and ELN genes; inhibit enzyme synthesis for collagen and elastin degradation through ELANE, MMP1, MMP13, TIMP1 genes; does not affect DHFa cells, but does affect CHL-1 cells through the NRF2 gene; can stimulate DHFa cell proliferation through the FGF1 and FGF7 genes; has anti-inflammatory effects on DHFa cells, and has no effects on CHL-1 cells through the TGF-β gene. Ethanol M. tomentosa extract can suppress melanin production through the α-MSH and TYR genes. In summary, M. tomentosa can be used in human health and skin care products, the research was accomplished with the aim of deducing how more plant species with high bioactivities can be utilized without toxicity, the leaves of M. tomentosa can be easier to use correctly than fruits depending on the season and smell.

Keywords: Collagen, Elastin, α-EG, Morinda species, Pinoresinol, qRT-PCR