Pallidal activity related to posture and movement during reaching in the cat.

We tested the hypothesis that the pallidum contributes to the control of both posture and movement. We recorded neuronal activity from the pallidum in a task in which male cats reached forward from a standing posture to depress a lever. In agreement with previous studies, we found that a majority of pallidal cells (91/116 78%), including neurons in both the entopeduncular nucleus and the globus pallidus, showed significant modulations of their activity during reaching with the contralateral limb. Mostly different populations of cells were active during the transport (flexion) and lever press (extension) phase of the task. Most cells showed dynamic patterns of activity related to the movement. However, a modest proportion of modulated cells (18/91, 20%) showed properties consistent with a contribution to the control of anticipatory postural responses while a further 10% showed activity consistent with a contribution to postural support during the movement. While some cells that showed modified activity only during reaches with the contralateral forelimb, many cells (65/91, 71%) were also activated during reaches with the ipsilateral forelimb. This was particularly true for cells related to the lever press, many of which discharged similarly during reaches of either limb. This suggests a context-dependent control of movement and posture in which the same muscles are used for different functions during contralateral and ipsilateral reach. Comparison with the results from recordings made previously from the motor cortex and the pontomedullary reticular formation in the same task show more similarities with the former than the latter. Significance statement: Pathological changes in basal ganglia function frequently lead to problems with postural stability and gait initiation. Here we show that some neurons in one of the output regions of the basal ganglia, the pallidum, show discharge activity compatible with a contribution to postural control. At the same time, we note that such cells are a minority in this region with most cells being related to movement, rather than posture. We also show that many neurons are active during movements of both the contralateral and ipsilateral limbs, sometimes with identical discharge patterns. We suggest that this indicates a context-dependent regulation of movement and posture in the pallidum.

A secondary motor area contributing to interlimb coordination during visually guided locomotion in the cat.

We investigated the contribution of cytoarchitectonic cortical area 4δc, in the caudal bank of the cruciate sulcus of the cat, to the control of visually guided locomotion. To do so, we recorded the activity of 114 neurons in 4δc while cats walked on a treadmill and stepped over an obstacle that advanced toward them. A total of 84/114 (74%) cells were task-related and 68/84 (81%) of these cells showed significant modulation of their discharge frequency when the contralateral limbs were the first to step over the obstacle. These latter cells included a substantial proportion (27/68 40%) that discharged between the passage of the contralateral forelimb and the contralateral hindlimb over the obstacle, suggesting a contribution of this area to interlimb coordination. We further compared the discharge in area 4δc with the activity patterns of cells in the rostral division of the same cytoarchitectonic area (4δr), which has been suggested to be a separate functional region. Despite some differences in the patterns of activity in the 2 subdivisions, we suggest that activity in each is compatible with a contribution to interlimb coordination and that they should be considered as a single functional area that contributes to both forelimb-forelimb and forelimb-hindlimb coordination.

Bartha-K61 vaccine protects nursery pigs against challenge with novel european and asian strains of suid herpesvirus 1.

The present study investigates the pathogenicity of two recently isolated strains of Suid herpesvirus 1 (SuHV1), the Greek strain Hercules and the Chinese strain HeN1, in unvaccinated pigs and in pigs vaccinated with a Bartha-K61 strain. In an experiment performed in negative pressure kiosks (isolators), 45-day old seronegative pigs previously oronasally /intramuscularly vaccinated with the Bartha-K61 vaccine strain, along with unvaccinated controls, were challenged either with the Hercules strain or the HeN1 strain of SuHV1. All animals were observed daily for clinical signs and body temperature and nasal swabs, faeces, blood and bodyweight were collected up to a maximum period of 20 days post-challenge (dpc). The results showed that, in the unvaccinated pigs, HeN1 strain was more virulent than the Hercules strain, with increased mortality, shorter time to death and higher group clinical score (p < 0.05). However, after vaccination with the Bartha-K61 vaccine, there was a drastic reduction in morbidity, mortality, bodyweight loss and virus excretion to almost a similar extent in both strains (p < 0.05). No significant differences were seen among the pigs of the two vaccinated groups compared to unvaccinated unchallenged controls, except a slight elevation in body temperature and in clinical score in the HeN1 vaccinees at 2 and 3 dpc, while bodyweight gain was similar to that of the negative control pigs. Our study showed that despite differences in virulence, the standard vaccination scheme with the Bartha-K61 strain could equally protect nursery pigs against both the European and Chinese strains.

Microstimulation of the Premotor Cortex of the Cat Produces Phase-Dependent Changes in Locomotor Activity.

To determine the functional organization of premotor areas in the cat pericruciate cortex we applied intracortical microstimulation (ICMS) within multiple cytoarchitectonically identified subregions of areas 4 and 6 in the awake cat, both at rest and during treadmill walking. ICMS in most premotor areas evoked clear twitch responses in the limbs and/or head at rest. During locomotion, these same areas produced phase-dependent modifications of muscle activity. ICMS in the primary motor cortex (area 4γ) produced large phase-dependent responses, mostly restricted to the contralateral forelimb or hindlimb. Stimulation in premotor areas also produced phase-dependent responses that, in some cases, were as large as those evoked from area 4γ. However, responses from premotor areas had more widespread effects on multiple limbs, including the ipsilateral limbs, than did stimulation in 4γ. During locomotion, responses in both forelimb and hindlimb muscles were evoked from cytoarchitectonic areas 4γ, 4δ, 6aα, and 6aγ. However, the prevalence of effects in a given limb varied from one area to another. The results suggest that premotor areas may contribute to the production, modification, and coordination of activity in the limbs during locomotion and may be particularly pertinent during modifications of gait.

Contribution of the Entopeduncular Nucleus and the Globus Pallidus to the Control of Locomotion and Visually Guided Gait Modifications in the Cat.

We tested the hypothesis that the entopeduncular (EP) nucleus (feline equivalent of the primate GPi) and the globus pallidus (GPe) contribute to both the planning and execution of locomotion and voluntary gait modifications in the cat. We recorded from 414 cells distributed throughout these two nuclei (referred to together as the pallidum) while cats walked on a treadmill and stepped over an obstacle that advanced towards them. Neuronal activity in many cells in both structures was modulated on a step-by-step basis during unobstructed locomotion and was modified in the step over the obstacle. On a population basis, the most frequently observed change, in both the EP and the GPe, was an increase in activity prior to and/or during the swing phase of the step over the obstacle by the contralateral forelimb, when it was the first limb to pass over the obstacle. Our results support a contribution of the pallidum, in concert with cortical structures, to the control of both the planning and the execution of the gait modifications. We discuss the results in the context of current models of pallidal action on thalamic activity, including the possibility that cells in the EP with increased activity may sculpt thalamo-cortical activity.

The effect of temperature on persistence of SARS-CoV-2 on common surfaces.

BACKGROUND: The rate at which COVID-19 has spread throughout the globe has been alarming. While the role of fomite transmission is not yet fully understood, precise data on the environmental stability of SARS-CoV-2 is required to determine the risks of fomite transmission from contaminated surfaces. METHODS: This study measured the survival rates of infectious SARS-CoV-2, suspended in a standard ASTM E2197 matrix, on several common surface types. All experiments were carried out in the dark, to negate any effects of UV light. Inoculated surfaces were incubated at 20 °C, 30 °C and 40 °C and sampled at various time points. RESULTS: Survival rates of SARS-CoV-2 were determined at different temperatures and D-values, Z-values and half-life were calculated. We obtained half lives of between 1.7 and 2.7 days at 20 °C, reducing to a few hours when temperature was elevated to 40 °C. With initial viral loads broadly equivalent to the highest titres excreted by infectious patients, viable virus was isolated for up to 28 days at 20 °C from common surfaces such as glass, stainless steel and both paper and polymer banknotes. Conversely, infectious virus survived less than 24 h at 40 °C on some surfaces. CONCLUSION: These findings demonstrate SARS-CoV-2 can remain infectious for significantly longer time periods than generally considered possible. These results could be used to inform improved risk mitigation procedures to prevent the fomite spread of COVID-19.

Premotor Cortex Provides a Substrate for the Temporal Transformation of Information During the Planning of Gait Modifications.

We tested the hypothesis that the premotor cortex (PMC) in the cat contributes to the planning and execution of visually guided gait modifications. We analyzed single unit activity from 136 cells localized within layer V of cytoarchitectonic areas 6iffu and that part of 4δ within the ventral bank of the cruciate sulcus while cats walked on a treadmill and stepped over an obstacle that advanced toward them. We found a rich variety of discharge patterns, ranging from limb-independent cells that discharged several steps in front of the obstacle to step-related cells that discharged either during steps over the obstacle or in the steps leading up to that step. We propose that this population of task-related cells within this region of the PMC contributes to the temporal evolution of a planning process that transforms global information of the presence of an obstacle into the precise spatio-temporal limb adjustment required to negotiate that obstacle.

Similar Motor Cortical Control Mechanisms for Precise Limb Control during Reaching and Locomotion.

Throughout the course of evolution there has been a parallel development of the complexity and flexibility of the nervous system and the skeletomuscular system that it controls. This development is particularly evident for the cerebral cortical areas and the transformation of the use of the upper limbs from a purely locomotor function to one including, or restricted to, reaching and grasping. This study addresses the issue of whether the control of reaching has involved the development of new cortical circuits or whether the same neurons are used to control both locomotion and reaching. We recorded the activity of pyramidal tract neurons in the motor cortex of the cat both during voluntary gait modifications and during reaching. All cells showed generally similar patterns of activity in both tasks. More specifically, we showed that, in many cases, cells maintained a constant temporal relationship to the activity of synergistic muscle groups in each task. In addition, in some cells the relationship between the intensity of the cell discharge activity and the magnitude of the EMG activity was equally constant during gait modifications and reaching. As such, the results are compatible with the hypothesis that the corticospinal circuits used to control reaching evolved from those used to precisely modify gait.

Comparative analysis of cytokine transcript profiles within mediastinal lymph node compartments of pigs after infection with porcine reproductive and respiratory syndrome genotype 1 strains differing in pathogenicity.

Porcine reproductive and respiratory syndrome virus (PRRSV) induces a weak immune response enabling it to persist in different organs of infected pigs. This has been attributed to the ability of PRRSV to influence the induction of cytokine responses. In this study, we investigated the cytokine transcriptional profiles in different compartments of the mediastinal lymph node of pigs infected with three genotype 1 PRRSV strains of differing pathogenicity: the low virulence prototype Lelystad virus (LV), and UK field strain 215-06 and the highly virulent subtype 3 SU1-Bel isolate from Belarus. We have used a combination of laser capture micro-dissection (LCM) followed by real time quantitative PCR (RT-qPCR) and immunohistochemical (IHC) detection of immune cell markers (CD3, CD79a and MAC387) and RT-qPCR quantification of PRRSV and cytokine transcripts. Compared to mock infected pigs, we found a significant downregulation of TNF-α and IFN-α in follicular and interfollicular areas of the mediastinal lymph node from 3 days post-infection (dpi) in animals infected with all three strains. This was accompanied by a transient B cell depletion and T cell and macrophage infiltration in the follicles together with T cell depletion in the interfollicular areas. A delayed upregulation of IFN-γ and IL-23p19 was observed mainly in the follicles. The PRRSV load was higher in all areas and time-points studied in the animals infected with the SU1-Bel strain. This paper describes the first application of LCM to study the cytokine transcript profiles and virus distribution in different compartments of the lymph node of pigs.

Differential modulation of descending signals from the reticulospinal system during reaching and locomotion.

We tested the hypothesis that the same spinal interneuronal pathways are activated by the reticulospinal system during locomotion and reaching. If such were the case, we expected that microstimulation within the pontomedullary reticular formation (PMRF) would evoke qualitatively similar responses in muscles active during both behaviors. To test this, we stimulated in 47 sites within the PMRF during both tasks. Stimulation during locomotion always produced a strongly phase-dependent, bilateral pattern of activity in which activity in muscles was generally facilitated or suppressed during one phase of activity (swing or stance) and was unaffected in the other. During reaching, stimulation generally activated the same muscles as during locomotion, although the modulation of the magnitude of the evoked responses was less limb dependent than during locomotion. An exception was found for some forelimb flexor muscles that were strongly facilitated by stimulation during the swing phase of locomotion but were not influenced by stimulation during the transport phase of the reach. We suggest that during locomotion the activity in interneuronal pathways mediating signals from the reticulospinal system is subject to strong modulation by the central pattern generator for locomotion. During reach, we suggest that, for most muscles, the same spinal interneuronal pathways are used to modify muscle activity but are not as strongly gated according to limb use as during locomotion. Finally, we propose that the command for movement during discrete voluntary movements suppresses the influence of the reticulospinal system on selected forelimb flexor muscles, possibly to enhance fractionated control of movement.

Cortical contribution to visuomotor coordination in locomotion and reaching.

One of the hallmarks of mammals is their ability to make precise visually guided limb movements to attain objects. This is best exemplified by the reach and grasp movements of primates, although it is not unique to this mammalian order. Precise, coordinated, visually guided movements are equally as important during locomotion in many mammalian species, especially in predators. In this context, vision is used to guide paw trajectory and placement. In this review we examine the contribution of the fronto-parietal network in the control of such movements. We suggest that this network is responsible for visuomotor coordination across behaviours and species. We further argue for analogies between cytoarchitectonically similar cortical areas in primates and cats.

At Least Three Doses of Leading Vaccines Essential for Neutralisation of SARS-CoV-2 Omicron Variant.

Plasma samples taken at different time points from donors who received either AstraZeneca (Vaxzevria) or Pfizer (Comirnaty) or Moderna (Spikevax) coronavirus disease-19 (COVID-19) vaccine were assessed in virus neutralization assays against Delta and Omicron variants of concern and a reference isolate (VIC31). With the Pfizer vaccine there was 6-8-fold reduction in 50% neutralizing antibody titres (NT50) against Delta and VIC31 at 6 months compared to 2 weeks after the second dose; followed by 25-fold increase at 2 weeks after the third dose. Neutralisation of Omicron was only consistently observed 2 weeks after the third dose, with most samples having titres below the limit of detection at earlier timepoints. Moderna results were similar to Pfizer at 2 weeks after the second dose, while the titres for AstraZeneca samples derived from older donors were 7-fold lower against VIC31 and below the limit of detection against Delta and Omicron. Age and gender were not found to significantly impact our results. These findings indicate that vaccine matching may be needed, and that at least a third dose of these vaccines is necessary to generate sufficient neutralising antibodies against emerging variants of concern, especially Omicron, amidst the challenges of ensuring vaccine equity worldwide.

Highly Thermotolerant SARS-CoV-2 Vaccine Elicits Neutralising Antibodies against Delta and Omicron in Mice.

As existing vaccines fail to completely prevent COVID-19 infections or community transmission, there is an unmet need for vaccines that can better combat SARS-CoV-2 variants of concern (VOC). We previously developed highly thermo-tolerant monomeric and trimeric receptor-binding domain derivatives that can withstand 100 °C for 90 min and 37 °C for four weeks and help eliminate cold-chain requirements. We show that mice immunised with these vaccine formulations elicit high titres of antibodies that neutralise SARS-CoV-2 variants VIC31 (with Spike: D614G mutation), Delta and Omicron (BA.1.1) VOC. Compared to VIC31, there was an average 14.4-fold reduction in neutralisation against BA.1.1 for the three monomeric antigen-adjuvant combinations and a 16.5-fold reduction for the three trimeric antigen-adjuvant combinations; the corresponding values against Delta were 2.5 and 3.0. Our findings suggest that monomeric formulations are suitable for upcoming Phase I human clinical trials and that there is potential for increasing the efficacy with vaccine matching to improve the responses against emerging variants. These findings are consistent with in silico modelling and AlphaFold predictions, which show that, while oligomeric presentation can be generally beneficial, it can make important epitopes inaccessible and also carries the risk of eliciting unwanted antibodies against the oligomerisation domain.

Characterisation and natural progression of SARS-CoV-2 infection in ferrets.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the infectious disease COVID-19, which has rapidly become an international pandemic with significant impact on healthcare systems and the global economy. To assist antiviral therapy and vaccine development efforts, we performed a natural history/time course study of SARS-CoV-2 infection in ferrets to characterise and assess the suitability of this animal model. Ten ferrets of each sex were challenged intranasally with 4.64 × 104 TCID50 of SARS-CoV-2 isolate Australia/VIC01/2020 and monitored for clinical disease signs, viral shedding, and tissues collected post-mortem for histopathological and virological assessment at set intervals. We found that SARS-CoV-2 replicated in the upper respiratory tract of ferrets with consistent viral shedding in nasal wash samples and oral swab samples up until day 9. Infectious SARS-CoV-2 was recovered from nasal washes, oral swabs, nasal turbinates, pharynx, and olfactory bulb samples within 3-7 days post-challenge; however, only viral RNA was detected by qRT-PCR in samples collected from the trachea, lung, and parts of the gastrointestinal tract. Viral antigen was seen exclusively in nasal epithelium and associated sloughed cells and draining lymph nodes upon immunohistochemical staining. Due to the absence of clinical signs after viral challenge, our ferret model is appropriate for studying asymptomatic SARS-CoV-2 infections and most suitable for use in vaccine efficacy studies.

In vitro characterisation of SARS-CoV-2 and susceptibility of domestic ferrets (Mustela putorius furo).

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is an emerging virus that has caused significant human morbidity and mortality since its detection in late 2019. With the rapid emergence has come an unprecedented programme of vaccine development with at least 300 candidates under development. Ferrets have proven to be an appropriate animal model for testing safety and efficacy of SARS-CoV-2 vaccines due to quantifiable virus shedding in nasal washes and oral swabs. Here, we outline our efforts early in the SARS-CoV-2 outbreak to propagate and characterize an Australian isolate of the virus in vitro and in an ex vivo model of human airway epithelium, as well as to demonstrate the susceptibility of domestic ferrets (Mustela putorius furo) to SARS-CoV-2 infection following intranasal challenge.

Contribution of cells in the posterior parietal cortex to the planning of visually guided locomotion in the cat: effects of temporary visual interruption.

In the present study, we determined whether cells in the posterior parietal cortex (PPC) may contribute to the planning of voluntary gait modifications in the absence of visual input. In two cats we recorded the responses of 41 neurons in layer V of the PPC that discharged in advance of the gait modification to a 900-ms interruption of visual information (visual occlusion). The cats continued to walk without interruption during the occlusion, which produced only minimal changes in step cycle duration and paw placement. Visual occlusion applied during the period of cell discharge was without significant effect on discharge frequency in 57% of cells. In the other cells, the visual occlusion produced either significant decreases (18%) or increases (21%) of discharge activity (in 1 cell there was both an increase and a decrease). The mean latency of the changes was 356 ms for decreases and 252 ms for increases. In most neurons, discharge frequency, when modified, returned to the same levels as during unoccluded locomotion when vision was restored. In some cells, there were significant changes in discharge activity after the restoration of vision; these were associated with corrections of gait. These results suggest that the PPC is more involved in the visuomotor transformations necessary to plan gait modifications than in continual sensory processing of visual information. We further propose that cells in the PPC contribute both to the planning of gait modifications on the basis of only intermittent visual sampling and to visually guided online corrections of gait.

Sequential activation of motor cortical neurons contributes to intralimb coordination during reaching in the cat by modulating muscle synergies.

We examined the contribution of the motor cortex to the control of intralimb coordination during reaching in the standing cat. We recorded the activity of 151 pyramidal tract neurons (PTNs) in the forelimb representation of three cats during a task in which the cat reached forward from a standing position to press a lever. We simultaneously recorded the activity of muscles in the contralateral forelimb acting around each of the major joints. Cell activity was recorded with and without the presence of an obstacle requiring a modification of limb trajectory. The majority of the PTNs (134/151, 89%) modulated their discharge activity at some period of the reach while 84/151 (56%) exhibited a significant peak or trough of activity as the limb was transported from its initial position to the lever. These phasic changes of activity were distributed sequentially throughout the transport phase. A cluster analysis of muscle activity in two of the cats showed the presence of five muscle synergies during this transport period. One of the synergies was related to the lift of the paw from the support surface, two to flexion of the limb and dorsiflexion of the paw, one to preparation for contact with the lever, and one to the transport of the entire limb forward; a sixth synergy was activated during the lever press. An analysis of the phase of cell activity with respect to the phase of activity of muscles selected to represent each of these synergies showed that different populations of PTNs were activated sequentially and coincidentally with each synergy. We suggest that this sequential activation of populations of PTNs is compatible with a contribution to the initiation and modulation of functionally distinct groups of synergistic muscles and ultimately serves to ensure the appropriate multiarticular, intralimb coordination of the limb during reaching.

Host inhibits replication of European porcine reproductive and respiratory syndrome virus in macrophages by altering differential regulation of type-I interferon transcriptional response.

Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease caused by a positive RNA strand arterivirus. PRRS virus (PRRSV) interacts primarily with lung macrophages. Little is known how the virus subverts the innate immune response to initiate its replication in alveolar macrophages. Large-scale transcriptional responses of macrophages with different levels of susceptibility to PRRSV infection were compared over 30 h of infection. This study demonstrates a rapid and intense host transcriptional remodelling during the early phase of the replication of the virus which correlates with transient repression of type-I interferon transcript as early as 8 h post-infection. These results support the suggestion from previous studies that host innate immune response inhibits replication of European porcine reproductive and respiratory syndrome virus in macrophages by altering differential regulation of type-I interferon transcriptional response.

ChAdOx1 nCoV-19 (AZD1222) vaccine candidate significantly reduces SARS-CoV-2 shedding in ferrets.

Vaccines against SARS-CoV-2 are likely to be critical in the management of the ongoing pandemic. A number of candidates are in Phase III human clinical trials, including ChAdOx1 nCoV-19 (AZD1222), a replication-deficient chimpanzee adenovirus-vectored vaccine candidate. In preclinical trials, the efficacy of ChAdOx1 nCoV-19 against SARS-CoV-2 challenge was evaluated in a ferret model of infection. Groups of ferrets received either prime-only or prime-boost administration of ChAdOx1 nCoV-19 via the intramuscular or intranasal route. All ChAdOx1 nCoV-19 administration combinations resulted in significant reductions in viral loads in nasal-wash and oral swab samples. No vaccine-associated adverse events were observed associated with the ChAdOx1 nCoV-19 candidate, with the data from this study suggesting it could be an effective and safe vaccine against COVID-19. Our study also indicates the potential for intranasal administration as a way to further improve the efficacy of this leading vaccine candidate.

A contribution of area 5 of the posterior parietal cortex to the planning of visually guided locomotion: limb-specific and limb-independent effects.

We tested the hypothesis that area 5 of the posterior parietal cortex (PPC) contributes to the planning of visually guided gait modifications. We recorded 121 neurons from the PPC of two cats during a task in which cats needed to process visual input to step over obstacles attached to a moving treadmill belt. During unobstructed locomotion, 64/121 (53%) of cells showed rhythmic activity. During steps over the obstacles, 102/121 (84%) of cells showed a significant change of their activity. Of these, 46/102 were unmodulated during the control task. We divided the 102 task-related cells into two groups on the basis of their discharge when the limb contralateral to the recording site was the first to pass over the obstacle. One group (41/102) was characterized by a brief, phasic discharge as the lead forelimb passed over the obstacle (Step-related cells). These cells were recorded primarily from area 5a. The other group (61/102) showed a progressive increase in activity prior to the onset of the swing phase in the modified limb and frequently diverged from control at least one step cycle before the gait modification (Step-advanced cells). Most of these cells were recorded in area 5b. In both groups, some cells maintained a fixed relationship to the activity of the contralateral forelimb regardless of which limb was the first to pass over the obstacle (limb-specific cells), whereas others changed their phase of activity so that they were always related to activity of the first limb to pass over the obstacle, either contralateral or ipsilateral (limb-independent cells). Limb-independent cells were more common among the Step-advanced cell population. We suggest that both populations of cells contribute to the gait modification and that the discharge characteristics of the Step-advanced cells are compatible with a contribution to the planning of the gait modification.