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A study was carried out to determine the distribution and antimicrobial susceptibility profiles of Salmonella serovars from chickens from large-scale commercial (LSC), small-scale commercial (SSC), and rural free-range (RFR) farms of Zimbabwe. Pooled cloacal swabs were collected for culture and isolation of Salmonella spp. A chi-square test was used to assess distribution differences of salmonellas among the farming sectors. Approximately 10% (283/2833) of the swabs were positive for Salmonella enterica, with only subspecies enterica (98.6%) and arizonae (1.4%) being detected. The prevalence of S. enterica varied significantly (P<0.05) among areas, with Harare (27.8%) and Buhera (1.3%) recording the highest and the least prevalence, respectively. S. enterica was only isolated from LSC and SSC farms, with the former having a significantly (P<0.001; χ2=155.3) higher prevalence than the latter. S. arizonae was only isolated from the SSC farms while none were obtained from the RFR farms. The serovars isolated were Salmonella Enteritidis (72.8%), Group C (20.1%), Group B (4.2%), Salmonella Typhimurium (1.1%) and Salmonella Gallinarum (0.4%). S. Enteritidis predominated in the urban/periurban areas. Approximately 26% (53/206) of S. Enteritidis isolates were resistant to one or more antimicrobial agents. Resistance to tetracycline was the most common, while no resistance was detected for furazolidone, neomycin and trimethoprim-sulfamethoxazole. There were 12.1% multi-drug-resistant S. Enteritidis isolates, and the resistance to ampicillin/kanamycin was predominant. The identification of multi-drug-resistant S. Enteritidis is of public health concern. Thus, stringent control of S. Enteritidis will reduce the public health risk of human salmonellosis. [ABSTRACT FROM PUBLISHER]

A cross-sectional study was conducted to detect the presence of Salmonella spp., Candida albicans, Aspergillus spp., and antimicrobial residues in raw milk (n = 120) and processed cow milk (n = 20) from smallholder dairy farms from three sites in Zimbabwe. Culture and isolation of Salmonella spp., C. albicans, and Aspergillus spp. were performed using selective media, while antimicrobial residues were detected by a dye reduction test. No Salmonella, but C. albicans (17.5%; 21/120), Aspergillus spp. (0.8%; 1/120), and antimicrobial residues (2.5%; 3/120) were detected from raw milk. C. albicans was isolated from all three sites, while Aspergillus spp. and antimicrobial residues were detected from sites 1 and 3, respectively. From processed milk, only C. albicans (5%) was isolated while Aspergillus spp. and antimicrobial residues were not detected. These results suggested low prevalence of Salmonella spp. and Aspergillus spp. and a relatively high prevalence of C. albicans in raw milk from the smallholder farms. The potential public health risks of C. albicans and the detected antimicrobial residues need to be considered. Thus, educating farmers on improving milking hygiene and storage of milk and establishing programmes for monitoring antimicrobial residues may help to improve the safety of milk from smallholder farms. [ABSTRACT FROM AUTHOR]

Responses to tuberculin in Zebu cattle of the transhumant pastoral farming system in Karamoja region and Nakasongola district in the north-eastern and mid-central regions in Uganda, respectively, were investigated using a comparative intradermal tuberculin skin test. Of the 1864 cattle tested from 30 large units (superherds) in Karamoja and 7 herds in Nakasongola, a total of 28 animals from 19 herds (51.4%) tested positive. Inter-district tuberculin reactor prevalence variations seemed to be influenced by climate, with impact on both the management patterns and transmissibility of agent. High herd tuberculin reactor prevalence (51.4%) was attributed to widespread contacts and mixing of animals between herds. Low individual animal tuberculin test positivity (mean = 1.4%) was attributed to low transmissibility of the agent under the Karamoja climate, which is semi-arid, and to increased resistance due to non-specific response to environmental mycobacteria and natural selection, since there was no active control against bovine tuberculosis. Owing to similarities in management practices in Karamoja and widespread risk factors, it was difficult to identify which were more important, but variations in sources of drinking water pointed to provision of lake and borehole water during dry season as reducing the risk. Positive bovine tuberculin reactor prevalence and skin reactor status were related to age. [ABSTRACT FROM AUTHOR]

A cross-sectional study was performed in the livestock–wildlife interface areas of Lochinvar and Blue Lagoon National Parks and the non-interface area of Kazungula to determine the prevalence of antibodies to Brucella spp. in domestic ruminants and identify individual animal risk factors of infection. A total of 1245 cattle from 124 herds and 280 goats and sheep from 29 flocks were tested sequentially for Brucella antibodies using the Rose Bengal test (RBT) and competitive ELISA. In cattle, individual seroprevalence ranged from 14.1% to 28.1%, while herd sero–prevalence ranged from 46.2% to 74.0% in the three study areas. No goat or sheep tested positive for Brucella antibodies. Three types of cattle grazing strategies were encountered: locally grazed herds (LGH), transhumantly grazed herds (TGH) and river flood plain grazed herds (FGH). Brucella seroprevalence was seen to vary according to area and grazing strategy: Lochinvar and transhumant grazed herds recorded the highest figures, respectively. Age, sex and history of abortion were found to have independent effects on individual seroprevalence. This study establishes that brucellosis is endemic in domestic animals in the livestock–wildlife interface areas of Blue Lagoon and Lochinvar national parks and the disease is also present in Kazungula. We observed that type of grazing strategy had significant impact on cattle Brucella seroprevalence and that transhumant herds were at high risk of being infected. [ABSTRACT FROM AUTHOR]

United Nations (UN) sustainable development goal 6 focuses on clean water and sanitation for all. To date, one-third of the world’s population lack access to clean, safe water. The traditional water purification methods cannot monitor and control water quality parameters from the early stages. In this paper, Internet of Things (IoT) is used to develop a real-time water quality monitoring and control system for a water treatment plant. A microcontroller is developed and interfaced with sensors to detect any deviations in the water quality parameters from the World Health Organization (WHO) standards. Every stage of the water treatment process is tested for quality, and deviations from the WHO standard triggers an autonomous control measure. A simulation model and a working prototype are developed. The research produces the expected results: accurate monitoring and control of water parameters. [ABSTRACT FROM AUTHOR]

Selective laser melting (SLM) is one of the most well-known additive manufacturing methods available for the fabrication of functional parts from metal powders. Although SLM is now an established metal additive manufacturing technique, its widespread application in industry is still hindered by inherent phenomena, one of which is high residual stresses. Some of the effects of residual stresses–such as warping and thermal stress-related cracking–cannot be corrected by post processing. Therefore, establishing input process parameter combinations that result in the least residual stress magnitudes and related distortions and/or cracking is critical. This paper presents the influence of laser power, scanning speed, and layer thickness on residual stresses, distortions and achievable density for maraging steel 300 steel parts in order to establish the most optimum input parameter combinations. An analysis of the interdependence between process outcomes shows that high residual stress magnitudes lead to high dimensional distortions in the finished parts, whilst porous parts suffer relatively lower residual stresses and associated distortions. [ABSTRACT FROM AUTHOR]

Quality management has ceased to be an operational extra, and has become imperative to doing business in a saturated market environment with demanding clients. It is now a significant component of holistic operational management. The ultimate aim remains customer satisfaction, using the most effective techniques to ensure the most efficient and costeffective process. Statistical process control (SPC) and statistical quality control have been widely used in quality management to enhance process performance by reducing process variation. However, SPC can only be effective if implemented with a structured problem-solving tool. The diagnostic ability of statistics, coupled with a failure modes and effect analysis (FMEA) as the problem-solving methodology during an out-ofcontrol action plan, creates an environment conducive to improving processes and empowering employees. The failure modes and effect analysis was used to identify and eliminate sources of variation, which led to a reduction in process variation by 63 per cent and a decrease in defects by 160 961 per million opportunities. [ABSTRACT FROM AUTHOR]

Monitoring the presence of commensal and pathogenic respiratory microorganisms is of critical global importance. However, community-based surveillance is difficult because nasopharyngeal swabs are uncomfortable and painful for a wide age range of participants. We designed a methodology for minimally invasive self-sampling at home and assessed its use for longitudinal monitoring of the oral, nasal and hand microbiota of adults and children within families. Healthy families with two adults and up to three children, living in and near Liverpool, United Kingdom, self-collected saliva, nasal lining fluid using synthetic absorptive matrices and hand swabs at home every two weeks for six months. Questionnaires were used to collect demographic and epidemiological data and assess feasibility and acceptability. Participants were invited to take part in an exit interview. Thirty-three families completed the study. Sampling using our approach was acceptable to 25/33 (76%) families, as sampling was fast (76%), easy (76%) and painless (60%). Saliva and hand sampling was acceptable to all participants of any age, whereas nasal sampling was accepted mostly by adults and children older than 5 years. Multi-niche self-sampling at home can be used by adults and children for longitudinal surveillance of respiratory microorganisms, providing key data for design of future studies. [ABSTRACT FROM AUTHOR]

Longitudinal, community-based sampling is important for understanding prevalence and transmission of respiratory pathogens. Using a minimally invasive sampling method, the FAMILY Micro study monitored the oral, nasal and hand microbiota of families for 6 months. Here, we explore participant experiences and opinions. A mixed methods approach was utilised. A quantitative questionnaire was completed after every sampling timepoint to report levels of discomfort and pain, as well as time taken to collect samples. Participants were also invited to discuss their experiences in a qualitative structured exit interview. We received questionnaires from 36 families. Most adults and children >5y experienced no pain (94% and 70%) and little discomfort (73% and 47% no discomfort) regardless of sample type, whereas children ≤5y experienced variable levels of pain and discomfort (48% no pain but 14% hurts even more, whole lot or worst; 38% no discomfort but 33% moderate, severe, or extreme discomfort). The time taken for saliva and hand sampling decreased over the study. We conducted interviews with 24 families. Families found the sampling method straightforward, and adults and children >5y preferred nasal sampling using a synthetic absorptive matrix over nasopharyngeal swabs. It remained challenging for families to fit sampling into their busy schedules. Adequate fridge/freezer space and regular sample pick-ups were found to be important factors for feasibility. Messaging apps proved extremely effective for engaging with participants. Our findings provide key information to inform the design of future studies, specifically that self-sampling at home using minimally invasive procedures is feasible in a family context. [ABSTRACT FROM AUTHOR]

In this paper, a decision-making framework for implementing the concept of digitalisation within the South African tool, die, and mould-making (TDM) industry context is developed and employed. The purpose of this framework is to answer the following question: "Which digital technologies currently available on the market can be employed to improve the efficiency of shop-floor operations in the South African TDM industry?" An exhaustive literature study of existing digital technologies is conducted. Thereafter, TDM processes requiring digitalisation are identified through knowledge engineering. Based on the system requirements analysis, digital technologies relevant to the South African TDM context are then proposed. [ABSTRACT FROM AUTHOR]

Digitalisation has been advocated as a possible strategy to improve the competitiveness of tool, die, and mould-making (TDM) companies in the 21st century. The recent rise of digital technologies, such as Internet of Things devices, now makes digitalisation an achievable reality. This paper focuses on the digitalisation of shop-floor operations in the South African TDM industry through the development of a novel mobile data collection (MDC) tool known as a shop-floor management system (SMS). The developed SMS was deployed to, and validated in, a selected tooling company for various products. The developed system improved the shop-floor's real-time data collection. [ABSTRACT FROM AUTHOR]

Background: Studies have linked bats to outbreaks of viral diseases in human populations such as SARS-CoV-1 and MERS-CoV and the ongoing SARS-CoV-2 pandemic. Methods: We carried out a longitudinal survey from August 2020 to July 2021 at two sites in Zimbabwe with bat–human interactions: Magweto cave and Chirundu farm. A total of 1732 and 1866 individual bat fecal samples were collected, respectively. Coronaviruses and bat species were amplified using PCR systems. Results: Analysis of the coronavirus sequences revealed a high genetic diversity, and we identified different sub-viral groups in the Alphacoronavirus and Betacoronavirus genus. The established sub-viral groups fell within the described Alphacoronavirus sub-genera: Decacovirus, Duvinacovirus, Rhinacovirus, Setracovirus and Minunacovirus and for Betacoronavirus sub-genera: Sarbecoviruses, Merbecovirus and Hibecovirus. Our results showed an overall proportion for CoV positive PCR tests of 23.7% at Chirundu site and 16.5% and 38.9% at Magweto site for insectivorous bats and Macronycteris gigas, respectively. Conclusions: The higher risk of bat coronavirus exposure for humans was found in December to March in relation to higher viral shedding peaks of coronaviruses in the parturition, lactation and weaning months of the bat populations at both sites. We also highlight the need to further document viral infectious risk in human/domestic animal populations surrounding bat habitats in Zimbabwe. [ABSTRACT FROM AUTHOR]

Global interest in phytogenic feed additives as alternatives to antibiotics in feed has been spurred by the banning of antibiotic growth promoters by several countries. Suitable plant extracts for development of phytogenic feed additives should have therapeutic value and should also be safe. The aim of this study was to evaluate the antibacterial, antioxidant and anti-lipoxygenase activities as well as cytotoxicity of selected plant species used in poultry ethnomedicine in Zimbabwe. Antibacterial activity was determined against three ATCC strains (Staphylococcus aureus, Escherichia coli, Salmonella Enteritidis) and two clinical strains isolated from chickens (Escherichia coli and Salmonella Gallinarum) using a two-fold serial microdilution assay. Qualitative antibacterial bioautography was also carried out using the ATCC strains. Antioxidant activities of crude acetone and methanol extracts were determined using free radical scavenging assays whilst anti-lipoxygenase activity was evaluated using a ferrous oxidation-xylenol orange (FOX) assay. Cytotoxicity was evaluated using a tetrazolium-based colorimetric assay (MTT assay) on Vero monkey kidney cells. Erythrina abyssinica had the best antibacterial activity against both ATCC strains and clinical strains with minimum inhibitory concentration (MIC) values ranging from 0.02 to 0.156 mg/ml. Aloe greatheadii, Adenia gummifera (leaves), Senna singueana and Aloe chabaudii had moderate activity against the poultry pathogens. Bioautography showed that all ten plant species have antibacterial activity against the tested microorganisms with E. abyssinica and S. singueana having prominent bands of inhibition against both Gram-negative and Gram-positive bacteria. The acetone extract of S. singueana and the methanol extract of Euphorbia matabelensis had the most powerful antioxidant activities with mean IC 50 values of 1.43 μg/ml and 1.31 μg/ml respectively in the ABTS assay which were comparable with those of the positive controls (ascorbic acid and trolox). Bobgunnia madagascariensis, A. chabaudii , E. abyssinica and Tridactyle bicaudata extracts had reasonable antioxidant activity. The S. singueana extract had the most potent anti-lipoxygenase activity with a mean IC 50 value of 1.72 μg/ml. The cytotoxicity results showed that only the acetone extracts of A. greatheadii and S. singueana were relatively safe at concentrations that were active against the tested microorganisms (selective index >1). Regarding anti-lipoxygenase activity, extracts of B. madagascariensis , S. singueana, T. bicaudata and E. matabelensis were more active than toxic (selective index >5) indicating anti-inflammatory potential. This study showed that S. singueana had a cocktail of therapeutic activity and supports further investigation of this plant species for development of phytogenic poultry feed additives. Other plant species with noteworthy biological activities include B. madagascariensis, E. abyssinica, A. greatheadii, T. bicaudata and E. matabelensis. [Display omitted] • Several plant species are used in poultry ethnomedicine in Zimbabwe. • Extracts of selected plants had promising antibacterial activity against chicken pathogens. • Good selectivity index values and other useful bioactivities encourage further research. • Plants may be a useful source of antibiotic feed additive alternatives in poultry. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the sensitivity (Se) and specificity (Sp) of the Rose Bengal test (RBT), complement fixation test (CFT), the serum lateral flow assay (LFA serum) and the blood lateral flow assay (LFA blood) for the detection of antibodies to Brucella spp. using Bayesian latent class models (BLCMs). Sera and whole blood were collected from naturally infected cattle reared in smallholder, small-scale commercial and large-scale commercial farms in Zimbabwe (n = 1022) and Botswana (n = 770). The BLCMs were fitted under the assumption that conditional dependences existed between the tests. Based on the conditional dependence model, the RBT had the highest Se of 0.897 (95 % Probability Intervals: 0.854; 0.932) compared to 0.827 (0.773; 0.872), 0.812 (0.76; 0.858) and 0.809 (0.785; 0.832) for the LFA serum , LFA blood and CFT, respectively. The CFT recorded a higher Sp of 0.999 (0.995; 1.000) than the LFA serum 0.996 (0.99; 1.000), the LFA blood 0.984 (0.976; 0.991) and the RBT 0.969 (0.959; 0.978). The data indicated that both the Se and Sp of RBT and CFT and the Sp of LFA serum and LFA blood were conditionally independent, while the Se appeared to be conditionally dependent. These results indicated that none of the evaluated tests had perfect Se and Sp and consequently could not be used alone for the diagnosis of brucellosis in cattle from the studied farming sectors. Thus, based on high Se and Sp , respectively, a brucellosis testing regimen using the RBT (screening) and the LFA (confirmatory) may be considered. [ABSTRACT FROM AUTHOR]

Barrier bednets (BBnets), regular bednets with a vertical insecticidal panel to target mosquitoes above the bednet roof, where they are most active, have the potential to improve existing Insecticidal Treated Bednets (ITNs), by reducing the quantity of insecticide required per net, reducing the toxic risks to those using the net, potentially increasing insecticide choice. We evaluated the performance of PermaNet 3.0 (P3) and untreated (Ut) bed nets with and without pyrethroid and piperonyl butoxide roof barriers in killing pyrethroid-resistant and susceptible Anopheles gambiae, simultaneously video-recording mosquito flight tracks. Bioassay results showed that treated roof barriers, particularly the longitudinal P3 barrier (P3L) could be an effective addition to a bed net: P3 + P3L were consistently significantly more effective than the reference P3 bednet while performance of untreated nets could be raised to equal that of the reference P3 following the addition of a P3 barrier. The BBnet's potential to augment existing bednets and enhance their performance is considered., (© 2023. The Author(s).)

Background: Insecticide-treated nets (ITNs) using pyrethroids have been the main vector control tools deployed in malaria endemic countries and are responsible for the dramatic reduction in African malaria cases in the early 2000s. The World Health Organization (WHO) cone test was designed to assess the rapid toxicity effects of pyrethroid exposure on mosquito vectors but has yielded no insights beyond 60-min knockdown and 24-h mortality. As dual-active-ingredient (AI) ITNs become more widespread, bioassays that can provide realistic assessment of single- and dual-treated ITNs (i.e. nets with more than one active ingredient) are urgently needed., Methods: We present an augmentation of the cone test that enables accurate quantification of vector behavioural responses (specifically movement, spatial and temporal occupancy) to ITNs using video recording and bespoke software that uses background segmentation methods to detect spatial changes in the movement of mosquitoes within the cone. Four strains of Anopheles gambiae sensu lato (s.l.) were exposed to four ITNs (PermaNet 2.0, PermaNet 3.0, Olyset Net, Interceptor G2) and untreated nets in these modified cone tests. Life history data (post-exposure blood-feeding, blood meal weight, longevity) for individual mosquitoes were recorded., Results: All mosquitoes responded to the presence of ITNs, spending from 1.48 to 3.67 times more time in the upper region of the cone, depending on the ITN type. Of all ITNs, PermaNet 2.0 provoked the smallest change in behavioural response. Activity in the cone influenced observed post-exposure longevity, and in resistant strains exposed to Interceptor G2, the higher the activity, the greater the risk of dying, as long as the proportion of activity at the net surface was less than 50%. All ITNs inhibited blood-feeding, and smaller blood meals were taken when mosquitoes fed., Conclusions: The additional mosquito behaviour data obtained by using this modification to the WHO cone test provides unique insight into the innate responses of different mosquito strains on untreated nets and the entomological mode of action of ITNs, important evidence when evaluating ITN characteristics., (© 2023. The Author(s).)

Vector control interventions play a fundamental role in the control and elimination of vector-borne diseases. The evaluation of vector control products relies on bioassays, laboratory and semi-field tests using live insects to assess the product's effectiveness. Bioassay method development requires a rigorous validation process to ensure that relevant methods are used to capture appropriate entomological endpoints which accurately and precisely describe likely efficacy against disease vectors as well as product characteristics within the manufacturing tolerance ranges for insecticide content specified by the World Health Organization. Currently, there are no standardized guidelines for bioassay method validation in vector control. This report presents a framework for bioassay validation that draws on accepted validation processes from the chemical and healthcare fields and which can be applied for evaluating bioassays and semi-field tests in vector control. The validation process has been categorized into four stages: preliminary development; feasibility experiments; internal validation, and external validation. A properly validated method combined with an appropriate experimental design and data analyses that account for both the variability of the method and the product is needed to generate reliable estimates of product efficacy to ensure that at-risk communities have timely access to safe and reliable vector control products., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Many mosquito vectors rest inside human habitations, a behavioral trait that is exploited for vector control by indoor residual spraying (IRS) of interior walls with insecticide. Although IRS and its refined version targeted IRS are very effective against Aedes aegypti , they are expensive and logistically challenging to deliver in densely populated urban areas where outbreaks of dengue and other arboviruses are the greatest challenge. In experiments in Recife, Brazil, we set out to quantify the indoor resting behavior of Ae. aegypti at a level beyond that previously reported. We found that significantly more Ae. aegypti males, unfed and fed females visited the base of walls (height 0-20 cm, corresponding to 12.3% of the total wall surface) more frequently than upper wall areas, with the difference more pronounced at higher temperatures. When the lowest 20 cm of the walls was treated with an appropriate insecticide and colored black, we recorded up to 85% cumulative mortality after 24-h exposure in the experimental room. The findings are significant because feasibly, householders could treat this small and accessible target zone manually, without the need for visits by costly IRS teams or equipment, reducing insecticide use and enabling communities to actively protect their own indoor environment., (© The Author(s) 2023. Published by Oxford University Press on behalf of National Academy of Sciences.)

Rationale: serotype 3 (SPN3) is a cause of invasive pneumococcal disease and associated with low carriage rates. Following the introduction of pediatric 13-valent pneumococcal conjugate vaccine (PCV13) programs, SPN3 declines are less than other vaccine serotypes and incidence has increased in some populations coincident with a shift in predominant circulating SPN3 clade, from I to II. A human challenge model provides an effective means for assessing the impact of PCV13 on SPN3 in the upper airway. Streptococcus pneumoniae serotype 3 (SPN3) is a cause of invasive pneumococcal disease and associated with low carriage rates. Following the introduction of pediatric 13-valent pneumococcal conjugate vaccine (PCV13) programs, SPN3 declines are less than other vaccine serotypes and incidence has increased in some populations coincident with a shift in predominant circulating SPN3 clade, from I to II. A human challenge model provides an effective means for assessing the impact of PCV13 on SPN3 in the upper airway. Objectives: In a human challenge study involving three well-characterized and antibiotic-sensitive SPN3 isolates (PFESP306 [clade Ia], PFESP231 [no clade], and PFESP505 [clade II]), inoculum doses (10,000, 20,000, 80,000, and 160,000 cfu/100 μl) were escalated until maximal colonization rates were achieved, with concurrent acceptable safety. Methods: Presence and density of experimental SPN3 nasopharyngeal colonization in nasal wash samples, assessed using microbiological culture and molecular methods, on Days 2, 7, and 14 postinoculation. A total of 96 healthy participants (median age 21, interquartile range 19-25) were inoculated ( Measurement and Main Results:  = 6-10 per dose group, 10 groups). Colonization rates ranged from 30.0-70.0% varying with dose and isolate. 30.0% (29/96) reported mild symptoms (82.8% [24/29] developed a sore throat); one developed otitis media requiring antibiotics. No serious adverse events occurred. n  = 6-10 per dose group, 10 groups). Colonization rates ranged from 30.0-70.0% varying with dose and isolate. 30.0% (29/96) reported mild symptoms (82.8% [24/29] developed a sore throat); one developed otitis media requiring antibiotics. No serious adverse events occurred. Conclusions: An SPN3 human challenge model is feasible and safe with comparable carriage rates to an established Serotype 6B human challenge model. SPN3 carriage may cause mild upper respiratory symptoms.

Background: The WHO cone test is one of three tests currently used to evaluate the efficacy of insecticide-treated bed nets (ITNs). It generates two test outputs, knockdown and 24-h mortality, both indicative of immediate toxicity but that reveal little about the nature of mosquito and ITN interaction or how results translate to real-world settings., Methods: A human arm held 5 mm behind the net surface acted as a host attractant during cone tests and a smartphone was used to capture mosquito behaviour in the cone. Post-exposure blood feeding and survival for nine days were recorded; ingested blood meal size was determined by measuring excreted haematin. Four strains of Anopheles gambiae (insecticide susceptible: Kisumu and N'gousso; insecticide resistant: Banfora and VK7) were tested with and without the host attractant using untreated, Permanet 2.0 and Olyset nets. Video recordings were scan sampled every five seconds to record mosquito positions on either the net, in flight or in contact with the cone. Generalized estimating equations were used to analyse all data except survival within nine days which was analysed using Weighted Cox Regression., Results: Net contact was the most frequently recorded behaviour in all Anopheles spp. strains on all nets. Adding the human host as attractant triggered excitatory behaviours: in all strains, the magnitude of net contact was significantly decreased compared to tests without a host. ITN exposure altered the observed behaviour of the two susceptible strains, which exhibited a decreased response to the host during ITN tests. The resistant strains did not alter their behaviour during ITN tests. Significantly less net contact was observed during Olyset Net tests compared to Permanet 2.0. The host presence affected survival after exposure: Banfora and VK7 mosquitoes exposed to Permanet 2.0 with a host lived longer compared to tests performed without a host. However, mosquitoes that blood-fed and survived long enough to digest the blood meal did not exhibit significantly reduced longevity regardless of the presence of the host attractant., Conclusions: Simple modifications to the WHO cone test and extension of post-test monitoring beyond the current 24 h enable detailed behavioural characterizations of individual ITNs to be compiled. The effects observed from testing with a host and including blood feeding suggest that more representative estimates of true of ITN efficacy are gained with these modifications than when using the current testing protocol., (© 2022. The Author(s).)