Nguyen-Huan Pham-Khanh¹, Nhat-Quynh Huynh¹, Hong-Ngoc-Bao Le¹, Thi-Kim-Quy Ha^1*

¹College of Natural Sciences, Can Tho University, Campus II, 3/2 street, Ninh Kieu district, Can Tho City 94000, Vietnam

Abstract

The hazardous heavy metal ion Cr(VI) is harmful and easily mobile in the environment. Cr(VI) poisoning can cause delayed seed germination and damaged plant growth. This study suggested a green and simple synthesis method of ZnO microparticles (ZnO MPs) from Zn(CH₃COO)₂ solution and aqueous leaf extract of Dolichandrone spathacea (L. f.) K. Schum for protecting the legume plant (Vigna radiata) against the Cr(VI) stress. The optimized conditions for the synthesis of these MPs were determined using computational and experimental approaches. The characterization of ZnO MPs was analyzed by surface morphology, particle sizes, and elemental components using modern methods. The Zn MPs successfully exhibited the potential protective effects on the seed germination and seedling vigor of V. radiata under Cr(VI) stress. The results of PPR and ABTS assays also indicated that the antioxidant capacity of non-enzymatic antioxidants from leaves under Cr(VI) stress significantly reduced (46.83 ± 1.938% and 69.60 ± 2.17%, respectively) as compared to supplement of ZnO-MPs increased (55.44 ± 2.624% and 78.07 ± 0.820%, respectively). This study is an essential report for the agricultural field, which can apply further the new and green zinc-micronutrient fertilizer to mitigate the adverse effects of heavy metal contamination on crop cultivation.

Keywords: Green synthesis, Zinc oxide microparticles, Dolichandrone spathacea, Cr(VI) stress, Vigna radiata.

Wang Dongjing^1*, Su Zhonghua², Yuan Zhenjie¹, Mikhlid H. Almutairi³

¹Institute of Animal Husbandry and Veterinary, Tibet Autonomous Regional Academy of Agricultural Sciences, Lhasa 850009, China

²Tibet Autonomous Region Animal Disease prevention and Control Center, Lhasa 850009, China

³Zoology Department, College of Science, King Saud University, P.O. Box: 2455, 11451, Riyadh, Saudi Arabia

Abstract

This study was conducted to identify the biological characteristics of pathogen responsible for the death of Tibetan sheep in Nagarze County, Tibet, China. For this purpose, samples were collected from diseased animals and for bacterial culture and isolation. The isolated strains were subjected to several tests which included gram staining, biochemical identification, PCR amplification, genetic evolution analysis, in-vitro drug sensitivity and in-vivo pathogenicity tests. The results revealed that 7 strains of gram-positive cocci were isolated from Tibetan sheep, named TS-1, TS-2, TS-3, TS-4, TS-5, TS-6, and TS-7. These strains exhibited specific biochemical characteristics consistent with Enterococcus faecalis. Whereas, PCR amplification results were consistent with the expected outcomes on target band of approximately 1500 bp. Genetic evolutionary analysis revealed a significant homology (96.0%-99.9%) between the isolates and Enterococcus faecium. In-vitro drug sensitivity tests demonstrated that all the isolates exhibiting multiple drug resistance. Furthermore, the isolated strains displayed varying degrees of pathogenicity in mice. This study confirms that Enterococcus faecium is the causative agent for the deaths of Tibetan sheep. These findings enhance our understanding of the disease and suggest valuable insights for its prevention, control, and future research.

Keywords: Tibetan sheep, Enterococcus faecium, Isolation, Identification, Drug resistance, Pathogenicity

Muhammad Bilal¹, Shafia Tehseen Gul^1*, Muhammad Tariq Javed¹, Muhammad Saqib²

¹Department of Pathology, Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan

²Department of Clinical Medicine and Surgery, Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan

Abstract

Brucellosis is considered as a highly contagious and zoonotic disease globally and various diagnostic tests are available for its diagnosis. Keeping in view, the limitations of currently used serological techniques, a more precise, sensitive, and reliable loop mediated isothermal amplification (LAMP) assay was evaluated as an emerging diagnostic tool. In the current study, serum samples from cows (n=1989) and buffaloes (n=1467) were collected from the study area i.e., District Faisalabad and Toba Tek Singh in Punjab, Pakistan. As these two districts are present around the river Ravi in Punjab, Pakistan and known as the house of Nili Ravi breed. A number of dairy farms of local and imported cows are also present in this area. Initially the samples were screened by the RBPT and then subjected to c-ELISA for confirmation. Overall, 12.16 and 9.3% cows and buffaloes were seropositive through RBPT while 11.21 and 7.70% cows and buffaloes were seropositive via c-ELISA. The positive samples from c-ELISA were further subjected to molecular amplification at 1.5% agarose gel through LAMP assay. The current study concludes that LAMP assay is more sensitive as compared to other conventional PCR techniques while detecting true positives for brucellosis, so it can be used for confirmation of Brucella abortus as compared to PCR. In addition to the sensitivity and specificity and qualitative results can be observed through naked eye in LAMP, which is not possible in PCR.

Keywords: Brucellosis, LAMP, Quick diagnosis, Serological techniques, Rapid point of care

Hina Ali Ahmed¹, Nazeer Ahmed², Shahjahan Shabbir Ahmed², Shagufta Saddozai¹, Afroz Rais³,

Imran Ali Sani², Dawood Shahid², Shahbaz Khan^4*

¹Department of Zoology, Sardar Bahadur Khan Women’s University, Quetta, Pakistan

²Department of Biotechnology, Faculty of Life Sciences, BUITEMS, Quetta, Pakistan

³Department of Botany, Sardar Bahadur Khan Women’s University, Quetta, Pakistan

⁴Colorado Water Center, Colorado State University, Fort Collins, Colorado 80523, USA

Abstract

Deoxyribonucleic acid (DNA) sequencing is an emerging approach for revealing species diversity that has made species identification possible. This technique is an amazing and useful tool for studies in taxonomy, evolutionary biology, and biodiversity. Sequences of DNA can be used as genetic “barcodes” which could be used to  identify all animals, including insects. True flies, belonging to the order Diptera, are widely distributed and crucial components of ecosystems worldwide. Despite the rich biodiversity of Pakistan, our knowledge of various insect groups particularly flies remains limited. The current study was conducted in Quetta, Pakistan, from June 2017 to May 2018, to use DNA barcoding technique for the determination of the diversity of flies (658 bp sequence from the 5′-end of cytochrome oxidase I). Our analysis focused on a specific region of the cytochrome c oxidase 1 (COI) gene, known as the barcode region which provides valuable information for inferring evolutionary relationships and identifying species. The obtained sequences of 2,195 fly specimens were then compared and matched against the Barcode of Life Data System (BOLD), which assigned the specimens to 309 Barcode Index Numbers (BINs), which operates as a counterpart in the BOLD database. Among the families identified, Muscidae was the most dominant, with 283 specimens, followed by Cecidomyiidae with 184 specimens, and Ceratopogonidae with 164 specimens. A total 82 number of species were identified with Tricimba humeralis with the maximum catch.

Keywords: Arthropod, BOLD, Barcode index number, Biodiversity, Cytochrome c oxidase I, DNA barcoding

Mahmoud Abbas Ali¹, Mervat A. B. Mahmoud², Muhammad Shoaib³, Zeeshan Ahmad Bhutta^4*, Nada M. Ali⁵, Nadeem Ali⁶, Hani Z. Asfour⁷, Nisreen Rajeh⁸, Mohamed R. Eletmany^9,10

¹Department of Plant Protection, Faculty of Agriculture, South Valley University, Qena, Egypt

²Department of Zoology, Faculty of Science, South Valley University, Qena, Egypt

³Key Laboratory of New Animal Drug Project, Gansu Province/Key Laboratory of Veterinary Pharmaceutical Development, Ministry of Agriculture and Rural Affairs/Lanzhou Institute of Husbandry and Pharmaceutical Sciences of the Chinese Academy of Agricultural Sciences, Lanzhou 730050, China

⁴Laboratory of Veterinary Immunology and Biochemistry, College of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Republic of Korea

⁵Department of Chemistry, Faculty of Science, Al-Baha University, Al-Baha 65799,  Saudi Arabia

⁶Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah, 21589, Saudi Arabia

⁷Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah 21589, Saudi Arabia

⁸Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah, 21589, Saudi Arabia

⁹Department of Chemistry, Faculty of Science, South Valley University, Qena, Egypt

¹⁰TECS Department, Wilson College of Textiles, NC State University, Raleigh 27606, USA

Abstract

The red palm weevil (RPW), Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) is a major date palm pest. In this study, we aimed to isolate and identify the Serratia nematodiphila from RPW as potential biocontrol agents. We isolated the bacteria from infected RPW larvae and adults and identified using colony morphology characteristics, biochemical tests, and PCR followed by 16S rRNA sequencing. This is the first study reporting the Serratia nematodiphila as an extracellular symbiont of RPW from Egypt. The potential of this bacteria to be used as biocontrol agent was conducted by a screening bioassay through its effect on RPW eggs. The study noted that treated eggs were unable to hatch and not turned red in color, indicating the potential of this bacteria to be used as bio-pesticide. These results presented novel insights into the microbiome of RPW and suggest the potential of Serratia nematodiphila as a biocontrol agent for RPW management. Moreover, further studies are required to explore the mechanism and potential of these bacteria in field applications. Nevertheless, this study provides a promising direction for the development of sustainable and environmentally friendly RPW management strategies.

Keywords: Red Palm Weevils (RPW), Rhynchophorus ferrugineus, Coleoptera, Curculionidae, Serratia nematodiphila, Biocontrol agent

Yasir Bilal¹, Sabir Hussain^1*, Muhammad Shahid², Tanvir Shahzad¹, Faisal Mahmood¹

¹Department of Environmental Sciences, Government College University Faisalabad, Pakistan.

²Department of Bioinformatics & Biotechnology, Government College University Faisalabad, Pakistan

Abstract

Continuous discharge of textile wastewater consisting of variety of pollutants is a serious threat to ecosystems. Microbial bioremediation might serve as an effective approach for treating these unwanted contaminants. In this study, several bacteria isolated from textile wastewater were studied for decolorization of Congo red (CR) dye. The strain Alcaligenes sp. YB4 showed the most efficient potential to decolorize CR dye. Moreover, this strain efficiently decolorized CR while concurrently removing hexavalent chromium [Cr(VI)] in the same medium with maximum removal (> 90 %) of both pollutants at pH 7 and pH 8. The potential of YB4 for concurrent removal of both pollutants was observed to decrease with increasing concentration of NaCl. Similarly, Alcaligenes sp. YB4 efficiently removed the 91.6 % of CR and 95.7 % of Cr(VI) simultaneously, under static condition as compared to the shaking condition. While MS media amended with yeast extract showed about 92.2 % and 90.1 % removal of CR and Cr (VI) within 48 hours of incubation, respectively. Moreover, it was also noticed that presence of heavy metals effected the concurrent removal of both pollutants. The in-silico analysis of the azoreductase amplified from the strain YB4 identified the binding of CR with azoreductase and proposed the hypothesis that their association may be the primary cause of CR degradation. This study indicated that Alcaligenes sp. YB4, having azoreductase gene, is a potential resource to treat textile wastewater.

 Keywords: Congo red, Hexavalent chromium, Azoreductase, Molecular docking, Azo dyes