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Zika virus (ZIKV) epidemics with infections in pregnant women are associated with severe neurological disease in newborns. Although an arbovirus, ZIKV is also blood transfusion-transmitted (TT). Greater knowledge of the efficiency of ZIKV TT would aid decisions on testing and pathogen reduction technologies (PRT).Plasma units from ZIKV RNA-reactive blood donors were used to study infectivity in vitro, in mice, and in macaques. Furthermore, plasma units were subjected to PRT using amotosalen/ultraviolet light A (A/UVA) before transfusion.In vitro infectivity of ZIKV RNA-reactive plasma varied between 100 and 1000 international units (IU) of ZIKV RNA. Immunodeficient mice were more sensitive with as low as 32 IU sufficient to infect 50% of mice. 50-5500 IU of RNA led to TT in macaques using dose escalation of three different RNA-positive, seronegative plasma units. In contrast, RNA-reactive units collected postseroconversion were not infectious in macaques, even at a dose of 9 million IU RNA. After A/UVA PRT, transfusion of plasma containing up to 18 million IU was no longer infectious in vitro and did not result in ZIKV TT in macaques.Significant risks of ZIKV TT are likely confined to a relatively short viremic window before seroconversion, and that sensitive nucleic acid amplification testing likely identifies the majority of infectious plasma. PRT was demonstrated to be effective at preventing ZIKV TT. Considering that there is no approved ZIKV vaccine, these data are relevant to mitigate the risk of TT during the future ZIKV outbreaks. more...

An advance directive (AD) is a legal statement indicating the form of healthcare the patient would like to receive at a future time when he/she is no longer mentally competent to make decisions. As there are increased concerns to respect the preferences of medical treatment of patients suffering from terminal illnesses, AD receives more attention and recognition in healthcare settings. However, Hong Kong has neither statue nor direst case law on the legal status of AD. A public consultation was therefore launched by the government in 2019. The present study aimed to examine the views towards ADs in response to the proposed legislation.Fifty-nine responses were received from questionnaires and the findings formed a protocol and guideline for the focus groups. Twenty-three participants participated in two focus group discussions. A thematic analysis revealed three themes of views towards ADs legislations: public awareness, completion of ADs, and dying in place.Older and young participants held slightly different views on the acceptance of verbal and written format, and the presence of witnesses of ADs. Overall, participants considered the general public did not have adequate knowledge of AD.Findings suggested that public education should be enhanced, and guidance and protocols for healthcare professionals were needed after legislation of ADs. In addition, dying in place should also be facilitated while legislative barriers on Coroners Ordinance should be amended. more...

Estimation of fetal age is an important component of prenatal evaluation. The measure of the Crown-Rump Length (CRL) by ultrasound is one of the most frequently used methods to determine it. However, in certain pathologies, this measure could lose accuracy, and other measures should be taken to evaluate fetal age and to determine the normal growth of the conceptus. The main objective of this research was to compare different morphometric parameters with CRL in normal human fetuses to determine which of them has a stronger correlation with gestational age to be a useful measure when CRL could not be appropriately evaluated. We measured 10 different morphometric parameters and the weight in 120 human fetuses product of abortion that had externally normal morphology, and a Pearson´s Correlation test was made with each of the parameters with the gestational age determined using the CRL. Each of the 10 parameters had a significantly strong correlation with CRL. However, some of them have a stronger correlation and should be preferred when CRL is not available. If available, fetal age should be estimated using an ultrasound technique and measuring the CRL. Nevertheless, if an alteration in one of the structures affects its measure, a different parameter should be used. The limitations of each parameter should be noticed before using them. more...

The apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), was introduced from eastern North America into western North America via infested apples (Malus domestica Borkhausen) about 44 yr ago, where it subsequently adapted to 2 hawthorn species, Crataegus douglasii Lindley and C. monogyna Jacquin. Here, we test whether R. pomonella has also adapted to large-thorn hawthorn, Crataegus macracantha Loddiges ex Loudon, in Okanogan County, Washington State, USA. In 2020, 2021, and 2022, fruit of C. macracantha were shown to ripen in late September and were infested at rates from 0.7% to 3.0%. In laboratory rearing studies, large-thorn hawthorn flies from C. macracantha eclosed on average 9–19 days later than apple flies from earlier ripening apple (August–early September), consistent with large-thorn hawthorn flies having adapted to the later fruiting phenology of its host. In a laboratory no-choice test, significantly fewer (64.8%) large-thorn hawthorn than apple flies visited apples. In choice tests, greater percentages of large-thorn hawthorn than apple flies resided on and oviposited into C. macracantha versus apple fruit. Large-thorn hawthorn flies were also smaller in size than apple flies. Our results provide further support for the recursive adaptation hypothesis that R. pomonella has rapidly and independently specialized phenologically and behaviorally to different novel hawthorn hosts since its introduction into the Pacific Northwest of the USA, potentially leading to host race formation. more...

Seasonal temperatures select for eclosion timing of temperate insects and their parasitoids. In western North America, the fruit fly Rhagoletis zephyria Snow (Diptera: Tephritidae) is parasitized by the hymenopterous wasps Utetes lectoides (Gahan), an egg parasite, and Opius downesi Gahan, a larval parasite (both Braconidae). Eclosion of wasps should be timed with the presence of susceptible fly stages, but reports indicate U. lectoides ecloses in the absence of flies under no-chill conditions. Based on this, we tested the hypotheses that chill durations and no-chill temperatures both differentially regulate eclosion times of R. zephyria and its parasitic wasps. When fly puparia were chilled at ~3°C for 130–180 d, U. lectoides and O. downesi always eclosed on average later than flies. However, after 180-d chill, flies eclosed on average earlier than after 130- and 150-d chill, whereas eclosion times of U. lectoides and O. downesi were less or not affected by chill duration. When fly puparia were exposed to 20–22°C (no chill), U. lectoides eclosed before flies, with 88.9% of U. lectoides versus only 0.61% of flies eclosing. Taken together, findings show that eclosion times of flies are more sensitive to changes in chill duration than those of wasps. Flies are less sensitive than wasps to no-chill in that most flies do not respond by eclosing after no-chill while most wasps do. Our results suggest that shorter winters and longer summers due to climate change could cause mismatches in eclosion times of flies and wasps, with potentially significant evolutionary consequences. more...

Organic yard waste from western Washington, U.S. that may contain puparia of apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), had been moved to central Washington for composting, threatening the $3 billion apple industry concentrated in that region. Heating waste to kill fly puparia before it is transported could be a solution to this problem. Here, we report results of studies in 2016–2021 that sought to identify a minimum heat treatment simulating that obtained using a low-pressure steam generator for maximizing kill of R. pomonella puparia. In two experiments, puparia were exposed to temperatures ramped linearly over 6 h from 21°C to 47.8, 51.1, 55.0, or 60.0°C in an oven. The 47.8, 51.1, and 55°C treatments did not achieve 100% mortality, although only one adult fly from 4,000 puparia was found in the 55°C treatment, while no puparia survived the 60°C treatment. In a third, similar experiment, no puparia out of 2,400 exposed to 55°C survived. In a fourth and final experiment conducted over 3 years, no puparia out of 61,223 exposed to a 6-h ramp from 21°C to 55°C followed by a 1-h hold time at 55°C produced flies. In addition, all puparia in this treatment died. Based on 42.3 to 69.8% control survival, 31,217 puparia were killed by this treatment with no survivors, for a probit 8.7190 level of security. Results suggest that the 55°C and 1-h hold time treatment here is close to the minimum heat regime needed for disinfesting organic waste of R. pomonella puparia. more...

The U.S. Food and Drug Administration only gave emergency use authorization of the BNT162b2 and mRNA-1273 SARS-CoV-2 vaccines for infants 6 months and older in June 2022. Yet questions regarding the durability of vaccine efficacy, especially against emerging variants, in this age group remain. We demonstrated previously that a two-dose regimen of stabilized prefusion Washington SARS-CoV-2 S-2P spike (S) protein encoded by mRNA encapsulated in lipid nanoparticles (mRNA-LNP) or purified S-2P mixed with 3M-052, a synthetic Toll-like receptor (TLR) 7/8 agonist, in a squalene emulsion (Protein+3M-052-SE) was safe and immunogenic in infant rhesus macaques. Here, we demonstrate that broadly neutralizing and spike-binding antibodies against variants of concern (VOCs), as well as T cell responses, persisted for 12 months. At 1 year, corresponding to human toddler age, we challenged vaccinated rhesus macaques and age-matched nonvaccinated controls intranasally and intratracheally with a high dose of heterologous SARS-CoV-2 B.1.617.2 (Delta). Seven of eight control rhesus macaques exhibited severe interstitial pneumonia and high virus replication in the upper and lower respiratory tract. In contrast, vaccinated rhesus macaques had faster viral clearance with mild to no pneumonia. Neutralizing and binding antibody responses to the B.1.617.2 variant at the day of challenge correlated with lung pathology and reduced virus replication. Overall, the Protein+3M-052-SE vaccine provided superior protection to the mRNA-LNP vaccine, emphasizing opportunities for optimization of current vaccine platforms. The observed efficacy of both vaccines 1 year after vaccination supports the implementation of an early-life SARS-CoV-2 vaccine. more...

Tolerance of terrestrial insects in temperate regions to water immersion and hypoxia has rarely been studied but can be an important adaptation to moist environments, with implications for insect dispersal through waterways. In the Pacific Northwest of the United States, apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), can be found in riparian habitats subject to flooding. Here, survival of R. pomonella larvae and different age puparia after flotation or immersion in 13.3°C or 21.1°C water for 1–12 d was determined. Larvae sank in water and when submerged for 1 or 2 d suffered greater mortality than control larvae. Fewer young (1–2 d old) than older puparia (13–15 d old) floated in water. When immersed in water for 1–12 d, young puparia suffered greater mortality than older puparia, which were not affected by water immersion. Consequently, fewer adult flies eclosed from puparia that had been water treated when young than older. Adult flies from pre-chill and post-chill puparia that had been water treated eclosed later than control flies, but treatment flies survived about 60 d and reproduced. Although newly-formed puparia are susceptible to hypoxic water conditions, increased buoyancy and water tolerance occur rapidly after formation, perhaps making survival possible and allowing water-borne dispersal of older puparia. more...

Chimeric antigen receptor (CAR) T cells are ineffective against solid tumors with immunosuppressive microenvironments. To overcome suppression, we engineered circuits in which tumor-specific synNotch receptors locally induce production of the cytokine IL-2. These circuits potently enhance CAR T cell infiltration and clearance of immune-excluded tumors, without systemic toxicity. The most effective IL-2 induction circuit acts in an autocrine and T cell receptor (TCR)- or CAR-independent manner, bypassing suppression mechanisms including consumption of IL-2 or inhibition of TCR signaling. These engineered cells establish a foothold in the target tumors, with synthetic Notch–induced IL-2 production enabling initiation of CAR-mediated T cell expansion and cell killing. Thus, it is possible to reconstitute synthetic T cell circuits that activate the outputs ultimately required for an antitumor response, but in a manner that evades key points of tumor suppression. more...

Endosymbiont-induced cytoplasmic incompatibility (CI) may play an important role in arthropod speciation. However, whether CI consistently becomes associated or coupled with other host-related forms of reproductive isolation (RI) to impede the transfer of endosymbionts between hybridizing populations and further the divergence process remains an open question. Here, we show that varying degrees of pre- and postmating RI exist among allopatric populations of two interbreeding cherry-infesting tephritid fruit flies (Rhagoletis cingulata and R. indifferens) across North America. These flies display allochronic and sexual isolation among populations, as well as unidirectional reductions in egg hatch in hybrid crosses involving southwestern USA males. All populations are infected by a Wolbachia strain, wCin2, whereas a second strain, wCin3, only co-infects flies from the southwest USA and Mexico. Strain wCin3 is associated with a unique mitochondrial DNA haplotype and unidirectional postmating RI, implicating the strain as the cause of CI. When coupled with nonendosymbiont RI barriers, we estimate the strength of CI associated with wCin3 would not prevent the strain from introgressing from infected southwestern to uninfected populations elsewhere in the USA if populations were to come into secondary contact and hybridize. In contrast, cytoplasmic-nuclear coupling may impede the transfer of wCin3 if Mexican and USA populations were to come into contact. We discuss our results in the context of the general paucity of examples demonstrating stable Wolbachia hybrid zones and whether the spread of Wolbachia among taxa can be constrained in natural hybrid zones long enough for the endosymbiont to participate in speciation. more...

Triclosan (TCS), a synthetic antimicrobial, has been incorporated into a variety of personal care products worldwide. Since 2016, upon the recommendation of World Health Organization (WHO), the use of TCS-containing sutures (TCSCS) to prevent surgical site infections (SSI) has become a common practice. However, TCS is a subject of environmental and public health concern and has been ranked as a chemical of serious concern to be avoided based on GreenScreen® hazard assessment. Recent reports on the efficacy of TCSCS to prevent SSI have revealed conflicting results. Here, we aim to highlight and discuss various safety and antimicrobial resistance concerns associated with the generalized use of TCSCS. Further, we emphasize that the long-term health benefits must be evaluated before the incorporation of TCS into surgical sutures. more...

Sensing physical forces is a critical first step in mechano-transduction of cells. Zyxin, a LIM domain-containing protein, is recruited to force-bearing actin filaments and is thought to repair and strengthen them. Yet, the precise force-induced protein interactions surrounding zyxin remain unclear. Using BioID analysis, we identified proximal proteins surrounding zyxin under normal and force-bearing conditions by label-free mass spectrometry analysis. Under force-bearing conditions, increased biotinylation of α-actinin 1, α-actinin 4, and AFAP1 were detected, and these proteins accumulated along force-bearing actin fibers independently from zyxin, albeit at a lower intensity than zyxin. VASP also accumulated along force-bearing actin fibers in a zyxin-dependent manner, but the biotinylation of VASP remained constant regardless of force, supporting the model of a free zyxin-VASP complex in the cytoplasm being corecruited to tensed actin fibers. In addition, ARHGAP42, a RhoA GAP, was also identified as a proximal protein of zyxin and colocalized with zyxin along contractile actin bundles. The overexpression of ARHGAP42 reduced the rate of small wound closure, a zyxin-dependent process. These results demonstrate that the application of proximal biotinylation can resolve the proximity and composition of protein complexes as a function of force, which had not been possible with traditional biochemical analysis. more...

Even in patients whose tumours exhibit increased T cell infiltration, resistance to immune checkpoint inhibitor (ICI) therapy is common. We investigated mechanisms of ICI resistance using engineered mouse models with increased neoantigen burden and T cell infiltration. We found that in ICI-resistant tumours, T cells upregulate an NF-κB-driven inflammatory circuit in the tumour microenvironment (TME), culminating in PMN-MDSC recruitment and tumour escape. This resistance circuit was molecularly driven by IL-1, TNFα, and G-CSF, and their genetic or pharmacologic inhibition increased ICI efficacy. Furthermore, we identified a subset of human TNF-expressing T cells that expand following ICI therapy and are associated with increased tumour NF-κB signaling. These data reveal a surprising mechanism of ICI resistance whereby T cells instruct NF-κB-mediated inflammation to paradoxically drive a resistance circuit in participating tumours, refining our understanding of ICI response and resistance with important therapeutic implications. more...

How different macronutrients are utilized at various stages of pupal diapause and the effects of winter length on nutrient reserves remain poorly studied for most insects. Western cherry fruit fly, Rhagoletis indifferens (Diptera: Tephritidae), is a specialist on cherries in higher latitudes or elevations in western North America that exhibits a obligate pupal diapause requiring chilling before adult development can occur. We determined the relationship between metabolic reserves and diapause status in R. indifferens pupae, testing the hypotheses that lipids are the primary reserves utilized during diapause and that long periods of warmth deplete these reserves more than periods of cold. Effects of 0- to 20-week durations at 3°C and subsequent exposure to 23°C and 16:8 L:D (warm rearing conditions) for 0 to 7 weeks on lipid, protein, soluble carbohydrates, and glycogen reserves of R. indifferens pupae were determined. During diapause, lipid reserves were the primary source of energy utilized by R. indifferens, while protein and soluble carbohydrates levels were stable throughout diapause and thus less utilized. At post-diapause, glycogen levels fluctuated the most, indicating that lipid reserves were utilized to produce glycogen to support metabolism for adult fly development. Unchilled pupae did not deplete lipid reserves, unlike chilled pupae, likely because unchilled pupae remained in diapause. Rhagoletis indifferens may have evolved a nutrient utilization strategy typical of rigid diapausing insects in higher latitude environments. more...

Rhesus macaques are natural hosts to multiple viruses including rhesus cytomegalovirus (RhCMV), rhesus rhadinovirus (RRV), and Simian Foamy Virus (SFV). While viral infections are ubiquitous, viral transmissions to uninfected animals are incompletely defined. Management procedures of macaque colonies include cohorts that are Specific Pathogen Free (SPF). Greater understanding of viral transmission would augment SPF protocols. Moreover, vaccine/challenge studies of human viruses would be enhanced by leveraging transmission of macaque viruses to recapitulate expected challenges of human vaccine trials.This study characterizes viral transmissions to uninfected animals following inadvertent introduction of RhCMV/RRV/SFV-infected adults to a cohort of uninfected juveniles. Following co-housing with virus-positive adults, juveniles were serially evaluated for viral infection.Horizontal viral transmission was rapid and absolute, reaching 100% penetrance between 19 and 78 weeks.This study provides insights into viral natural histories with implications for colony management and modeling vaccine-mediated immune protection studies. more...

Breeding success should increase with prior knowledge of the surrounding environment, which is dependent upon an animal's ability to evaluate habitat. Prospecting for nesting locations and migratory stopover sites are well-established behaviours among bird species. We assessed whether three species of California dabbling ducks – mallards, Anas platyrhynchos, gadwall, Mareca strepera, and cinnamon teal, Spatula cyanoptera – in Suisun Marsh, California, U.S.A., a brackish marsh, prospect for suitable wetlands in the week prior to brooding. K-means cluster analyses grouped 29 mallard and gadwall hens into three groups. One group (N = 13) demonstrated evidence of brood site prospecting, with the fewest and latest prebrooding wetland visits. Of these hens, seven visited their future brood pond an average of 1.14 times and only shortly before brooding (1.29 days), obtaining current information on habitat suitability. For the remaining six hens, we did not detect a brooding wetland visit, possibly due to data limitations or because these hens acquired sufficient familiarity with the wetland habitat during nest breaks in adjacent wetlands, obviating the need to prospect the specific brood pond. The second identified group of hens (N = 11) visited the brooding wetland most frequently (on 4.55 days), further in advance (5.27 days), with the fewest unique wetland visits and the earliest brooding date (26 May). The final group of hens (N = 5) were the latest to brood (21 June) and visited the most wetlands, possibly due to less water or more broods present across the landscape. Brood ponds were always farther from the nest than the nearest ponds, indicating that habitat suitability or presence of conspecifics is more important to brood site selection. Prospecting provides hens with knowledge about current habitat conditions and allows them to ‘crowdsource’ public information regarding use of that habitat by other brooding hens. Prospecting may, therefore, benefit ducks inhabiting ephemeral habitats like those within Suisun Marsh, where brood habitat is limited and water cover changes rapidly during the breeding season. more...

Medical incidents in public hospitals in Hong Kong have persisted for years. The operating theatres are one of the places where medical accidents occur, especially affecting service quality. The surgical team is a substantial cause of medical incidents, possibly because of human mistakes, environmental, equipment, and system failures. Not all surgery departments will implement uniform working styles. The Hospital Authority may set a management plan to unify practice and to identity the problems faced. Therefore, the government and Hospital Authority should focus on human resources, especially in terms of professional training and retention of staff. After all, surgery is a task of teamwork. more...

BackgroundThe safety profile of antimicrobials used during pregnancy is one important consideration in the decision on how to treat and provide postexposure prophylaxis (PEP) for plague during pregnancy.MethodsWe searched 5 scientific literature databases for primary sources on the safety of 9 antimicrobials considered for plague during pregnancy (amikacin, gentamicin, plazomicin, streptomycin, tobramycin, chloramphenicol, doxycycline, sulfadiazine, and trimethoprim-sulfamethoxazole [TMP-SMX]) and abstracted data on maternal, pregnancy, and fetal/neonatal outcomes.ResultsOf 13 052 articles identified, 66 studies (case-control, case series, cohort, and randomized studies) and 96 case reports were included, totaling 27 751 prenatal exposures to amikacin (n = 9), gentamicin (n = 345), plazomicin (n = 0), streptomycin (n = 285), tobramycin (n = 43), chloramphenicol (n = 246), doxycycline (n = 2351), sulfadiazine (n = 870), and TMP-SMX (n = 23 602). Hearing or vestibular deficits were reported in 18/121 (15%) children and 17/109 (16%) pregnant women following prenatal streptomycin exposure. First trimester chloramphenicol exposure was associated with an elevated risk of an undescended testis (odds ratio [OR] 5.9, 95% confidence interval [CI] 1.2–28.7). Doxycycline was associated with cardiovascular malformations (OR 2.4, 95% CI 1.2–4.7) in 1 study and spontaneous abortion (OR 2.8, 95% CI 1.9–4.1) in a separate study. First trimester exposure to TMP-SMX was associated with increased risk of neural tube defects (pooled OR 2.5, 95% CI 1.4–4.3), spontaneous abortion (OR 3.5, 95% CI 2.3–5.6), preterm birth (OR 1.5, 95% CI 1.1–2.1), and small for gestational age (OR 1.6, 95% CI 1.2–2.2). No other statistically significant associations were reported.ConclusionsFor most antimicrobials reviewed, adverse maternal/fetal/neonatal outcomes were not observed consistently. Prenatal exposure to streptomycin and TMP-SMX was associated with select birth defects in some studies. Based on limited data, chloramphenicol and doxycycline may be associated with adverse pregnancy or neonatal outcomes; however, more data are needed to confirm these associations. Antimicrobials should be used for treatment and PEP of plague during pregnancy; the choice of antimicrobials may be influenced by these data as well as information about the risks of plague during pregnancy. more...

Western cherry fruit fly, Rhagoletis indifferens Curran, is a quarantine pest of sweet cherries in the Pacific Northwest of the United States that overwinters as diapausing pupae. Eclosion responses of R. indifferens puparia to different low temperature durations and postdiapause conditions affect the pest status of the fly. Here, we determined the effects of holding R. indifferens puparia at 3°C for 0, 1, 2, 5, 10, 15, and 20 wk on adult eclosion times and rates at two simulated temperate and two simulated tropical climate treatments over 40 wk. When puparia were chilled 0, 1, or 2 wk, adult eclosion across the four climate treatments displayed a bimodal distribution with low eclosion at 3 wk and high eclosion at 23–35 wk. When puparia were chilled ≤ 10 wk, there was a weaker bimodal distribution. However, when puparia were chilled 15–30 wk, eclosion was more synchronous and occurred at 5–7 wk across the four postchill climate treatments. Eclosion was greater at a postdiapause temperature of 26°C than 23°C. Timing to 50% eclosion was faster at longer photoperiod (16:8 L:D) than shorter (12:12 L:D). The bimodality of eclosion in respect to the duration of low temperature exposure may be indicative of univoltine insect species with obligate diapause that may span over two seasons. more...

Insecticides mixed in sugar-protein baits or insecticides alone have been used to control tephritid fruit flies for many years. Here, effects of cyantraniliprole, spinetoram, and the biopesticide Chromobacterium subtsugae extract in sucrose-yeast extract bait or alone on kill and oviposition of western cherry fruit fly, Rhagoletis indifferens Curran (Diptera: Tephritidae), were evaluated in the laboratory. Flies were exposed to dry insecticide bait or insecticide alone in the presence or absence of a nontoxic food strip. Spinetoram alone caused greater mortality than cyantraniliprole alone, while cyantraniliprole in bait caused mortality as high as spinetoram bait and greater mortality than cyantraniliprole alone. Chromobacterium subtsugae extract alone but not in bait caused significant mortality compared with controls, but was much less effective than cyantraniliprole and spinetoram. Spinetoram alone reduced oviposition more than cyantraniliprole alone. However, cyantraniliprole bait reduced oviposition as much or more than spinetoram alone or spinetoram bait. Cyantraniliprole and spinetoram baits caused greater mortality when a nontoxic food strip was absent than present, but there was no corresponding reduction in oviposition. Chromobacterium subtsugae extract did not significantly reduce oviposition compared with controls. Potential benefits of using cyantraniliprole baits as an additional or alternative method to using more toxic spinosyn insecticides for controlling R. indifferens warrant study. more...

In efforts to increase rigor and reproducibility, the USA National Primate Research Centers (NPRCs) have focused on qualification of reagents, cross-laboratory validations, and proficiency testing for methods to detect infectious agents and accompanying immune responses in nonhuman primates. The pathogen detection working group, comprised of laboratory scientists, colony managers, and leaders from the NPRCs, has championed the effort to produce testing that is reliable and consistent across laboratories. Through multi-year efforts with shared proficiency samples, testing percent agreement has increased from as low as 67.1% for SRV testing in 2010 to 92.1% in 2019. The 2019 average agreement for the four basic SPF agents improved to96% (86.5% BV, 98.9 SIV, 92.1 SRV, and 97.0 STLV). As new pathogens such as SARS coronavirus type 2 emerge, these steps can now be quickly replicated to develop and implement new assays that ensure rigor, reproducibly, and quality for NHP pathogen detection. more...