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Abstract Lead (Pb) is toxic to the development and growth of rice plants. Nanoparticles (NPs) have been considered one of the efficient remediation techniques to mitigate Pb stress in plants. Therefore, a study was carried out to examine the underlying mechanism of iron (Fe) and silicon (Si) nanoparticle-induced Pb toxicity alleviation in rice seedlings. Si–NPs (2.5 mM) and Fe-NPs (25 mg L−1) were applied alone and in combination to rice plants grown without (control; no Pb stress) and with (100 µM) Pb concentration. Our results revealed that Pb toxicity severely affected all rice growth-related traits, such as inhibited root fresh weight (42%), shoot length (24%), and chlorophyll b contents (26%). Moreover, a substantial amount of Pb was translocated to the above-ground parts of plants, which caused a disturbance in the antioxidative enzyme activities. However, the synergetic use of Fe- and Si–NPs reduced the Pb contents in the upper part of plants by 27%. It reduced the lethal impact of Pb on roots and shoots growth parameters by increasing shoot length (40%), shoot fresh weight (48%), and roots fresh weight (31%). Both Si and Fe–NPs synergistic application significantly elevated superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and glutathione (GSH) concentrations by 114%, 186%, 135%, and 151%, respectively, compared to plants subjected to Pb stress alone. The toxicity of Pb resulted in several cellular abnormalities and altered the expression levels of metal transporters and antioxidant genes. We conclude that the synergistic application of Si and Fe-NPs can be deemed favorable, environmentally promising, and cost-effective for reducing Pb deadliness in rice crops and reclaiming Pb-polluted soils.

Experimental research with animals can help the prevention, cure, and alleviation of human ailments. Animal research facilities are critical for scientific advancement, but they can also pose a higher risk than other biomedical laboratories. Zoonosis, allergic reactions, bites, cuts, and scratches by animals are all substantial concerns that can occur in animal facilities. Furthermore, human error and unexpected animal behavior pose a risk not just to humans, but also to the environment and the animals themselves. The majority of biosafety and biosecurity training programs focus on clinical and biomedical laboratories dealing with human safety factors, with little emphasis on animal biosafety. The current virtual training was designed to improve biosafety and biosecurity capabilities of animal laboratory personnel, researchers, and veterinarians from different regions of Pakistan. The results revealed that understanding was improved regarding triggers for risk assessment in addition to annual and regular reviews (56% to 69%), biosecurity (21% to 50%), decontamination (17% to 35%), safe handling of sharps (21% to 35%), Dual Use Research of Concern (DURC) (17% to 40%), Personal Protective Equipment (PPE) usage by waste handlers (60.9% to 75%), waste management (56% to 85%), animal biosafety levels (40.57% to 45%), and good microbiological practices and procedures (17% to 35%). To bring human and animal laboratories up to the same level in terms of biosafety and biosecurity, it is critical to focus on areas that have been overlooked in the past. Training programs focusing on animal biosafety should be conducted more frequently to strengthen bio risk management systems in animal research facilities.

Laboratory diagnostic capacity is crucial for an optimal national response to a public health emergency such as the COVID-19 pandemic. Preventing laboratory-acquired infections and the loss of critical human resources, especially during a public health emergency, requires laboratories to have a good biorisk management system in place. In this study, we aimed to evaluate laboratory biosafety and biosecurity in Pakistan during the COVID-19 pandemic. In this cross-sectional study, a self-rated anonymous questionnaire was distributed to laboratory professionals (LPs) working in clinical diagnostic laboratories, including laboratories performing polymerase chain reaction (PCR)-based COVID-19 diagnostic testing in Punjab, Sindh, Khyber Pakhtunkhwa, and Gilgit-Baltistan provinces as well as Islamabad during March 2020 to April 2020. The questionnaire assessed knowledge and perceptions of LPs, resource availability, and commitment by top management in these laboratories. In total, 58.6% of LPs performing COVID-19 testing reported that their laboratory did not conduct a biorisk assessment before starting COVID-19 testing in their facility. Only 31% of LPs were aware that COVID-19 testing could be performed at a biosafety level 2 laboratory, as per the World Health Organization interim biosafety guidelines. A sufficiently high percentage of LPs did not feel confident in their ability to handle COVID-19 samples (32.8%), spills (43.1%), or other accidents (32.8%). These findings demonstrate the need for effective biosafety program implementation, proper training, and establishing competency assessment methods. These findings also suggested that identifying and addressing gaps in existing biorisk management systems through sustainable interventions and preparing LPs for surge capacity is crucial to better address public health emergencies.

Introduction: The cyclical process of hazard identification, risk assessment, risk mitigation, and review is a key step in developing a biorisk management (BRM) system. This paper describes how this process was initiated in two laboratories in Pakistan using a unique model of blended learning. Methods: A training needs analysis showed that the staff had very little knowledge of BRM systems. A workshop using a unique blended model was conducted in which virtual and in-presence learning occurred simultaneously. This workshop aimed to train the participants by applying two key concepts from the World Health Organization Laboratory Biosafety Manual 4th edition: 1) the cyclical process of risk assessment and 2) mapping the core biorisk and establishing heightened control measures in the laboratories of the participants based on the risk assessment. All scenarios and examples used in the training were from the participants’ laboratory work processes. Results: Prior to this project, no risk assessment was conducted in these laboratories. After the workshop, a risk assessment was performed for six work processes. In addition, seven core requirements and three heightened control measures were mapped, a biorisk officer was appointed, and a biosafety committee was convened. Furthermore, a biorisk manual, a biological waste management plan, an occupational health center, and a system for audits and inspections are being developed. Discussion and conclusion: BRM training is not a one-time effort; it has to be strengthened to ensure the development and implementation of a comprehensive and sustainable BRM system. Training must be applicable to local settings and incremental, in a way that participants are not overloaded with information.

The increasing threat posed by state and nonstate actors seeking to misuse scientific and technological developments means it is more important than ever to ensure the responsible conduct of life sciences research, biosecurity research, and dual-use research of concern (DURC). Pakistan has a rapidly growing research environment and, being a signatory to the Biological Toxins and Weapons Convention, is committed to raising DURC awareness among its researchers. To this end, a series of five workshops were conducted in Pakistan between July 2018 and January 2020 with the support of the United States organization Health Security Partners. The workshops targeted universities offering postgraduate programs in life and medical sciences. The purpose of the workshops was to enable participants to understand the concept of DURC, review their own research, determine if any of their experiments may be DURC, identify potential risks while conducting the research, and develop protocols to mitigate DURC threats. A pre-and post-workshop questionnaire evaluated the knowledge of the participants. Evaluations of both the course and facilitators were obtained at each workshop.

IntroductionThe COVID-19 pandemic has resulted in enormous increases in laboratory activities to keep pace with diagnostic testing and research efforts. However, traditional training, technical assistance, and capacity-building approaches were disrupted by the travel and movement restrictions put in place to control the spread of the disease. To address the needs of laboratorians and managers to conduct laboratory activities safely and securely during the pandemic, a highly interactive virtual training (IVT) workshop on biorisk management during COVID-19 was conducted through active learning strategies that connected speakers with participants. The objective of the training was to increase the basic knowledge and standards of biosafety and biosecurity practices, risk assessment, and control measures with reference specifically to the context of the COVID-19 pandemic and apply a rigorous evaluation methodology to assess the effectiveness of the IVT. The training covered a broad range of topics and encompassed national to international guidelines.MethodsParticipants were selected through official channels at the national level, focusing on institutions within Pakistan. The sessions included lectures from international experts in biorisk management concepts, and incorporated poll questions as well as pre- and post-tests and feedback on the speakers' knowledge and presentation skills, to increase interactivity. The pre- and post-test comprised similar multiple-choice questions and provided to every participant to ascertain the impact of the training on awareness and knowledge of biorisk management topics and concepts, and results were compared using paired t-tests. For feedback on the speakers, participants were asked to submit their ratings measured on a five-point Likert scale. The reliability of the Likert scale was estimated using Cronbach's alpha. Analyses were performed using Microsoft Excel and SPSS version 23.ResultsIn total, 52 individuals from different laboratories across Pakistan and Pakistani students from abroad (China) as well participated in at least one session of the IVT. The participants' pre- and post-test scores showed a significant increase in knowledge and awareness (p < 0.001). The obtained Cronbach's alpha score was >0.8, indicating high reliability of the generated feedback on the IVT approach and speakers.ConclusionThe IVT on biosafety and biosecurity in the context of the COVID-19 pandemic proved beneficial for laboratory professionals and could be a useful model to continue in the future for raising awareness and knowledge.

The Good Microbiological Practices & Procedures (GMPP) is the most significant risk control measure as per the fourth edition of the WHO laboratory biosafety manual. Among GMPP, one of the best practices is hand washing. WHO and other public health agencies have published several guidance documents on hand washing, that describe closing the tap using a disposable paper towel/tissue paper at the end of hand washing as one of the critical steps. In resource-limited settings, where disposable paper towels cannot be provided at all times, the staff is left with ambiguous instructions on how to close the tap. In this paper, a modified hand washing method is documented that doesn't necessitate the use of disposable paper towels. In this method, both hands and faucets remain in contact with soap for at least 40–60 s. The method was validated by the use of Glo Germ. A survey questionnaire was also designed and conducted for the lab staff (n = 12) of the two laboratories, where this method was implemented, to assess whether this hand washing method brought any improvement in their hand washing practices and implementation. All (100%) of the survey respondents reported that this method of hand washing is more applicable and implementable than the WHO-recommended hand washing technique. Eighty three percentage reported that this modified method of hand washing raised their hand washing compliance. The authors suggest that this hand washing method can be used in resource-limited laboratory settings as an effective GMPP to ensure infection control.

Okra (Abelmoschus esculentus L. (Moench) plays a significant role in humans nutrition because its fresh leaves, stems, flowers, pods and seeds, are used for multiple purposes. The present study attempted to determine the spatial variations in biochemical attributes of osmoprotectants and the oxidative defense system of okra plants. Samples of soil and okra plants (leaves and fruits) were collected from three different locations: Faisalabad region-1 (7 JB-I), Faisalabad region-2 (7 JB-II) and Pindi Bhattian. Chlorophyll contents, glycine betaine (GB), ascorbic acid (AsA), total phenolics, hydrogen peroxide (H2O2), proline, and malondialdehyde (MDA) contents were analyzed in the leaves and fruits of okra plants. Soil analyses showed that pH, electrical conductivity (EC), phosphorus (P), potassium (K), iron (Fe), and saturation of soil were higher in Faisalabad region 2, while organic matter, sand, Zn, and Cu were higher in the Pindi Bhattian region. The results from okra leaves showed that Pindi Bhattian had higher chlorophyll a, GB and H2O2 contents, while Faisalabad region 1 had a higher ratio of chlorophyll a/b compared to the other regions. However, Faisalabad regions 2 and 1 had higher leaf phenolic contents, Faisalabad regions 1 and 2 showed higher leaf proline contents, and Faisalabad region 2 possessed higher AsA and MDA contents. Analyses of okra fruits showed that Faisalabad region 2 had higher chlorophyll a and total chlorophyll contents, while Faisalabad region 1 had higher chlorophyll b contents. Faisalabad region 2 and Pindi Bhattian had higher ratios of chlorophyll a/b, and Faisalabad region 1 showed higher phenolic, AsA, H2O2, and MDA contents of okra fruit, whereas the Faisalabad regions exhibited higher proline and GB contents than the Pindi Bhattian region. Overall, okra leaves and fruits showed better responses in the Faisalabad regions, and these results may be used to screen for okra cultivars with better tolerance under different environmental conditions.

Financial, cultural, and managerial hurdles have made biosafety and biosecurity measures difficult in resource-constrained countries like Pakistan. Because of increasing awareness of biorisk management, diagnostic and research laboratories have made major advances in biosafety and biosecurity in the recent decade. As a result, identifying and addressing gaps in biorisk management has never been more critical. The purpose of this study was to assess the current situation of personal protective equipment (PPE), biosafety behavior, waste management, biosafety and biosecurity measures, training and safety, and health services in diagnostic and research laboratories across Pakistan's Khyber Pakhtunkhwa (KP) province. We adapted the WHO Laboratory Assessment tool (2012) and CWA 15793 (Biorisk management guidelines) for conducting a cross-sectional survey, which was distributed among various laboratories in KP. The survey included 30 laboratories, including 11 diagnostic and 19 research laboratories. In comparison to diagnostic laboratories, biorisk management practices in research laboratories were better in terms of PPE, biosafety behavior, waste management, biosafety measures, biosecurity measures, trainings, and safety and health services. KP laboratories' biorisk management practices have improved over time, according to our findings. However, we were able to identify inadequacies that would require considerable improvements to the current setups based on the WHO and CWA 15793 recommendations. Organizations can tailor their biosafety measures and training to address identified gaps using the presented KP snapshot.

Dual-use research poses a significant challenge for scientists in the biomedical field and for global health security in general. As the scientific knowledge and materials required for the development of biological agents become progressively more accessible and inexpensive, there is an increased need to understand and improve the governance of scientific research. Prevention of the misuse of facilities, equipment, agents, and scientific knowledge requires high levels of awareness of the concept of dual-use research, starting with early-career scientists and graduate students. In this study, the attitudes and level of awareness of postgraduate students in Pakistan toward the issues surrounding dual-use research were assessed through a survey containing both quantitative and qualitative questions in 32 universities in 4 provinces, federal area, and Azad Jammu and Kashmir regions of Pakistan; 933 students responded. Most (58.2%) had never heard of dual-use research of concern (DURC), while 18.5% had heard the term but were unsure of its meaning. Irrespective of prior knowledge, a higher percentage of students (68.6%) felt an obligation to report research misuse. Considering the need for DURC training, 94.1% of the respondents agreed that the principal investigator should take the responsibility to train students on DURC at the start of a research project. When experimental results having dual-use potential, 69.1% indicated they would publish with limited protocol, with 43.5% indicating they would publish the limited protocol only if there was a way for scientists to access their data. The survey results revealed limited DURC awareness among researchers across Pakistan. However, the respondents, although not formally educated about DURC, were quite aware of its impact. The information gained in this survey will be valuable in addressing country-specific awareness and training needs.