Fayyaz Ahmad¹, Muhammad Shahid Mahmood^1*, Riaz Hussain^2*, Sajjad Ur Rahman¹, Rao Zahid Abbas³

¹Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan

²Department of Pathology, Faculty of Veterinary and Animal Sciences, Islamia University of Bahawalpur, Pakistan

³Department of Parasitology, Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan

Abstract

Hemorrhagic septicemia (HS) caused by Pasteurella multocida (P. multocida) is one of the leading causes of bovine respiratory problems in dairy animals. Therefore, identification of reliable and useful biomarkers of naturally occurring HS disease is of vital important for early diagnosis and monitoring of efficacy of treatment. Therefore, the current study was conducted to estimate the status of hematology, serum biochemistry, oxidative stress and antioxidant enzymes in dairy cattle (52) in and around district Bahawalpur. Deep nasal swabs and jugular blood was collected from normal and morbid animals showing signs of respiratory distress for molecular characterization of P. multocida. Different visceral tissues were obtained from animals died of respiratory signs for histopathological investigations. Results revealed significantly (P < 0.05) increased quantity of oxidative stress biomarkers while different antioxidant enzymes decreased significantly in erythrocyte of infected animals. Results on hematology revealed significantly increased total leukocyte counts and neutrophil counts while significantly decreased hemoglobin, hematocrit, monocyte, lymphocyte and erythrocyte counts. Results on serum biomarkers showed significantly (P < 0.05) increased quantity of different serum profile in P. multocida positive cattle. Grossly, lungs were congested, hyperemic and consolidated. Frothy exudate was observed in trachea of P. multocida infected cattle. Results on microscopic observation showed different pathological lesions in lungs, liver, heart and kidneys of P. multocida confirmed cases. Based on the results of our study, it can be suggested that continuous monitoring of disease is necessary to lower the prevalence of infectious agent. It is also suggested that blood biochemistry, oxidative stress and antioxidant oxidant enzymes are useful and reliable tools to clarify the pathogenesis of disease for proper therapeutics.

Keywords: Hemorrhagic septicemia (HS), Pasteurella multocida, Hemato-biochemistry, Oxidative stress, Antioxidant enzyme, Histopathology

Wenli Sun^1,2, Yu Wang^1,2, Jian Zhang^1,2, Yikai Yin^1,2, Mengqi Duan^1,2, Chamba Yangzom^1,2, Peng Shang^1,2*

¹College of Animal Science, Tibet Agriculture and Animal Husbandry University, Linzhi, Tibet, China

²The Provincial and Ministerial Co-founded Collaborative Innovation Center for R & D in Tibet Characteristic Agricultural and Animal Husbandry Resources, Tibet, China

Abstract

Tibetan pigs exhibit remarkable characteristics, particularly their heightened tendency for fat deposition and increased resistance to diseases in comparison to Yorkshire pigs. Surprisingly, there has been a noticeable scarcity of research delving into the underlying mechanisms responsible for these advantageous traits, especially from the perspective of intestinal microorganisms, particularly those inhabiting the duodenum. To bridge this research gap, the study harnessed 16S rRNA sequencing to conduct a comprehensive examination of the duodenal microbiota in Tibetan and Yorkshire pigs. The results stemming from amplicon sequencing of duodenal contents unveiled that at the phylum level, Firmicutes dominated the duodenal microbiota in both Tibetan and Yorkshire pigs. Intriguingly, Tibetan pigs showcased a significantly reduced proportion of Bacteroidetes in comparison to Yorkshire pigs (P < 0.05), leading to a substantially higher Firmicutes to Bacteroidetes ratio in Tibetan pigs (55.95) as opposed to Yorkshire pigs (3.86) (P < 0.05). Of remarkable note, at the genus level, Tibetan pigs displayed a significantly elevated relative abundance of Lactobacillus spp. when compared to Yorkshire pigs (P < 0.01). Functional predictions pertaining to the duodenal microbiota in both pig breeds primarily revolved around amino acid metabolism, cofactor and vitamin metabolism, terpene and polyketide metabolism, amino acid derivative metabolism, and lipid metabolism. This study underscores the intricate and interdependent relationship between the composition and abundance of the duodenal microbiota and the unique characteristics of fat deposition and disease resistance in Tibetan pigs. It suggests Lactobacillus spp. as significant contributors to fat deposition, the development of the duodenal intestinal barrier, and immune function. Additionally, the Firmicutes-to-Bacteroidetes ratio appears to be associated with fat deposition. These findings provide valuable insights that can serve as a valuable reference for future endeavors related to the development and utilization of Tibetan pigs.

Keywords: Tibetan pigs, Duodenum, Intestinal microorganisms, Fat deposition, Disease resistance

Ning Wang^1,2,3, Yuanlin Xiang¹, Yi Ma^1,3, Puyu Zhang¹, Xiaoli Zhou¹, Haoran Zhu¹, Zhi Zhang¹, Zaixin Li¹, Xiongjun Xiao¹, Manli He⁴, Muhammad Aamer Mehmood^1,5*, Hui Zhu^1,3*

 ¹ College of Bioengineering, Sichuan University of Science and Engineering, Zigong, China

² Luzhou Laojiao Co. Ltd, Luzhou, China

³ Engineering Technology Research Center of Special Grain for Wine Making, Yibin 644000, China

⁴ Laboratory Animal Center, Southwest Medical University, Luzhou, China

⁵ Department of Bioinformatics and Biotechnology, Government College University Faisalabad, Faisalabad 38000, Pakistan

Abstract

Spirulina platensis (S. platensis) produces a variety of biologically active compounds that exhibit antioxidative, anti-inflammatory, antibacterial, and immunoregulatory properties. Here, dextran sulfate sodium (DSS) was used to develop an animal model of ulcerative colitis (UC) to evaluate the potential protective benefits of fermented S. platensis against DSS-induced colitis in mice. Gut microbiota alterations were investigated using 16S ribosomal RNA (rRNA) gene sequencing. Real-Time Quantitative PCR (RT-qPCR) was used to detect the expression of mRNA of inflammatory factors and pathway-related molecules in the inflammatory process. The results showed that fermented S. platensis could reverse the DSS-induced weight loss and colon length shortening in mice. The study of the 16S rRNA sequence showed that treatment with fermented S. platensis changed the gut microbiota of mice, with an increase in the relative abundance of beneficial bacteria such as Lachnospira. According to RT-qPCR and histopathological analyses, fermented S. platensis also improved the loss of goblet cells and neutrophil infiltration induced by DSS, while improving anti-inflammatory capacity. In addition, compared with the model group, the fermentation group significantly downregulated the relative expression of MyD88/TLR4 signaling pathway genes compared with the non-fermentation group. Overall, this investigation demonstrated that fermentative S. platensis can reduce DSS-induced UC by regulating gut microbiota composition, and the MyD88/TLR4 signaling pathway.

Keywords: Spirulina platensis, Rhizopus oligosporus, Ulcerative colitis, Gut microbiota

Sobia Khaliq¹, Muhammad Asif Aziz^1*, Ata-ul-Mohsin¹, Ishfaq Ahmed Hafiz²

¹Department of Entomology, PMAS-Arid Agriculture University, Rawalpindi, Pakistan

²Department of Horticulture, PMAS-Arid Agriculture University, Rawalpindi, Pakistan

Abstract

The performance, life history, and parasitism rate of Diadegma insulare (Cresson) (Hymenoptera: Ichneumonidae) on Plutella xylostella (Linnaeus, 1857) (Lepidoptera: Plutellidae) reared on six different cruciferous cultivars (Broccoli, Cabbage, Canola, Cauliflower, Chinese cabbage and PakChoi) was evaluated under controlled conditions (25±2 °C, 65±5% RH, and 16 L: 8 D photoperiod). The life history parameters were compared using the age-stage and two-sex life table theory. Significant variations were observed in different life stages of D. insulare on its host, P. xylostella, reared on different cruciferous genotypes. The adult duration of D. insulare ranges from 7.70 day on Pak Choi to 6.65 day on Cabbage. The maximum (19.83 offspring) and minimum (9.23 offspring) net reproductive rates of D. insulare were observed on cultivars Pak Choi and Cabbage, respectively. The increase in intrinsic rate ranged from the highest (0.1970 day-¹) on Broccoli to lowest (0.1471 day-¹) on Cabbage. The maximum (0.2814 host) finite predation rate was recorded on Broccoli, and was the minimum (0.2112 hosts) on Cabbage. The values of stable predation rate increased from 0.1824 (host/ parasitoid) to 0.2308 (host/ parasitoid) with changes in host diet. All tested brassica cultivars affected the life performance of D. insulare. The results confirmed that D. insulare showed better performance on Broccoli genotype when compared to other tested genotypes. Hence, Broccoli can be used for the rearing of D. insulare on a large scale for mass releases of this parasitoid in the brassica fields to get effective suppression in the population of P. xylostella.

Keywords: Biological control, D. insulare, Demography, Intrinsic rate of increase, Cruciferous plants.

Dunia A. Al-Farraj¹, Muhammad Kashif^2*, Fateh Ullah², Hafiz Muhammad Ali³, Abdul Qayyum³, Asma Yamin⁴, Jawaria Aslam⁵, Shabana Bibi⁶, Sayed M. Eldin^7*, 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 12^th day while those treated with P. juliflora or Betaderm-N cream healed on 15^th day. The wounds treated with combination gel showed a significantly (p<0.05) higher healing rate and completely healed the wound by 9^th 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 7^th (p<0.05), 14^th (p<0.05) and 21^st (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

Muhammad Khalid¹, Najam us Sahar Sadaf Zaidi¹, Naeem Rashid², Muhammad Tahir^1*

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

Thitiwan Jumpa¹, Jutarop Phetcharaburanin², Manida Suksawat², Kunlaya Pattanagul³, Wattana Pattanagul^1*

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

Muftia Chairin Nissa¹, Dewi Qurrota A’yuni¹, Setia Budi Sasongko¹, Aji Prasetyaningrum¹, Mohamad Djaeni^1*, 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

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