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The aims of this study were to determine the prevalence and associated risk factors of bovine herpes virus type-4 (BoHV-4) infection and describe the genetic characteristics and predominant genotypes of the virus in Yolo and Tulare counties, California. A cross-sectional study involving multi-stage sampling technique was used. One hundred and forty-eight post-partum cows were enrolled from 11 dairy farms. Uterine/vaginal samples were collected and tested for BoHV-4 and other co-infecting viruses using real-time PCR. Data were analysed using multilevel logistic mixed-effect model. Phylogenetic analysis of 10 BoHV-4 isolates was conducted by targeting three open reading frames (ORF3, ORF8 and ORF 22) of the viral genome. The prevalence of BoHV-4 infection was 22.3% (33/148), while post-partum metritis was 33.8% (48/142). Strong association was found between BoHV-4 infection and lactation number, lactation stage and post-partum metritis. The odds of being positive for BoHV-4 infection were 6.47 times (95% CI 1.17–35.92; P < 0.05) and 6.79 times (95% CI 1.19–38.55; P < 0.05) higher for cows in the fourth and fifth lactation, respectively, compared with cows in the first lactation. BoHV-4 was 8.27 times more likely (95% CI 1.43–47.94; P < 0.05) among cows in the early stage of lactation (0–120 days) compared with those in late lactation (>240 days). Cows with post-partum metritis were 4.51 times (95% CI 1.27–16.02; P < 0.05) more likely to test positive for BoHV-4 infection compared with those without post-partum metritis. Phylogenetic analysis of BoHV-4 based on sequencing of glycoprotein and thymidine kinase (TK) genes revealed genetic variability of the virus with glycoprotein B genotype 1 and TK genotype 2 as being dominant genotypes. The reported high genetic variability of BoHV-4 indicates the possibility of co-infection with multiple genotypes.

The emergence of avian reovirus variant strains has caused negative effects in the poultry industry worldwide. Regardless of the efforts in molecular characterization and classification of these variants, information about the pathogenicity, transmissibility, and immunosuppression in chickens is limited. The genomes of two variant strains (A and B) and a classic S1133 strain (C) belonging to the same sigma C genotype 1 were compared. Additionally, these strains were used in a challenge experiment to evaluate inoculated and indirectly exposed specific-pathogen-free chickens. The whole-genome sequence analysis of the three strains revealed nucleotide identity differences in the L3, M2, and S1 genes. Strains A and B also showed homology differences in the S4 gene, despite having high homologies in all other genes. The in vivo challenge experiments showed that, whereas variant A induced high viral loads in tendons, hearts, and duodena of inoculated chickens, variant B induced high viral loads in indirectly exposed chickens. Likewise, histopathology reflected differences in the pathologic effects induced by these strains. For instance, the B and C strains induced more severe microscopic lesions compared with the A strain. Lymphoid depletion was more severe in bursas than in thymi, and inoculated birds were more affected than exposed birds. In conclusion, different pathologic outcomes in chickens were observed depending on the strain and transmission route. This study provides insights onto the relationship between pathogenicity and genomic composition of avian reoviruses.

A questionnaire was designed in order to gather information about bedding material and footbath preparation and maintenance in different productive units across the state of California.This information was used to plan two experiments. In the first experiment, we tested the effectiveness of footbaths in inactivating highly pathogenic (HP) and low pathogenic (LP) avian influenza viruses (AIVs) on rubber boots. Surprisingly, quaternary ammonia– and quaternary ammonia + glutaraldehyde–based footbaths were not able to eliminate live HPAIV (H5N8) and LPAIV (H6N2) particles on boots, while a chlorine-based granulated disinfectant was able to destroy the virus at contact. These results demonstrated the potential of AIV, particularly the HPAIV isolate, to persist even if exposed to disinfecting footbaths, and suggest that footbaths, as a single tool, are not capable of preventing pathogen introduction into commercial flocks. In the second experiment, we investigated the persistence of HPAIV (H5N8) and LPAIV (H6N2) in bedding material and feces obtained from turkey, broiler, and egg-layer commercial productive units. Samples were collected at different times after spiking the bedding materials and feces. Results showed that HPAIV (H5N8) was more persistent than LPAIV (H6N2) in layer feces and bedding material obtained from commercial broilers and turkeys. Live HPAIV particles persisted 96 hr, the last time point measured, in layer feces and less than 60 hr in broiler and turkey bedding. In contrast, LPAIV persisted less than 24 hr after being spiked in all the different substrates. Further research in biosecurity practices such as footbath preparation and maintenance and better understanding of the mechanism of the increased persistence of AIV is warranted in order to identify effective litter treatments that destroy live virus in bedding material.

Real-time brain-computer interfaces (BCIs) that decode electroencephalograms (EEG) during motor imagery (MI) are powerful adjuncts to rehabilitation after neurotrauma. Further, immersive virtual reality (VR) could complement BCIs by delivering visual and auditory sensory feedback (VR biofeedback) congruent to user’s MI, enabling task-oriented therapies. Yet, therapeutic outcomes rely on user’s proficiency in evoking MI to attain volitional BCI-commanded VR interaction. While previous studies have explored multi-session BCIs, we investigated the impact of longitudinal training on sensorimotor neuromodulation using BCI combined with VR-mediated externally-cued and self-paced lower-limb MI tasks. The EEG-based BCI was coupled with real-time VR biofeedback congruent with the MI task. Over multiple training sessions in laboratory conditions, five unimpaired individuals progressively learnt to improve control over their EEG during MI virtual walking, corresponding with increased BCI classification accuracy. Further, similar improvements were found with four individuals with chronic complete spinal cord injury (SCI) using the system in real-world neurorehabilitation settings. These findings demonstrate that unimpaired and SCI impaired individuals learnt to control their sensorimotor EEG associated with MI tasks through VR-mediated BCI training, which was associated with improved BCI classification accuracy. Our findings highlight the potential of VR-mediated BCIs in enhancing neuromodulation, providing a foundation for future rehabilitation therapies.

This study focuses on virus isolation of avian reoviruses from a tenosynovitis outbreak between September 2015 and June 2018, the molecular characterization of selected isolates based on partial S1 gene sequences, and the full genome characterization of seven isolates. A total of 265 reoviruses were detected and isolated, 83.3% from tendons and joints, 12.3% from the heart and 3.7% from intestines. Eighty five out of the 150 (56.6%) selected viruses for sequencing and characterization were successfully detected, amplified and sequenced. The characterized reoviruses grouped in six distinct genotypic clusters (GC1 to GC6). The most represented clusters were GC1 (51.8%) and GC6 (24.7%), followed by GC2 (12.9%) and GC4 (7.2%), and less frequent GC5 (2.4%) and GC3 (1.2%). A shift on cluster representation throughout time occurred. A reduction of GC1 and an increase of GC6 classified strains was noticed. The highest homologies to S1133 reovirus strain were detected in GC1 (~77%) while GC2 to GC6 homologies ranged between 58.5 and 54.1%. Over time these homologies have been maintained. Seven selected isolates were full genome sequenced. Results indicated that the L3, S1 and M2 genes, coding for proteins located in the virus capsid accounted for most of the variability of these viruses. The information generated in the present study helps the understanding of the epidemiology of reoviruses in California. In addition, provides insights on how other genes that are not commonly studied add variability to the reovirus genome.

Abstract Background Four sampling techniques commonly are used for antemortem identification of pathogens from cattle with bovine respiratory disease (BRD): the nasal swab (NS), guarded nasopharyngeal swab (NPS), bronchoalveolar lavage (BAL), and transtracheal wash (TTW). Agreement among these methods has not been well characterized. Objective To evaluate agreement among TTW and NS, NPS, or BAL for identification of viral and bacterial pathogens in dairy calves with BRD. Animals One hundred dairy calves with naturally acquired BRD. Methods Calves were sampled by all 4 methods. Viral agents were identified by real-time RT-PCR, bacteria were identified by aerobic culture, and Mycoplasma bovis (M. bovis) isolates were speciated by PCR. Agreement among TTW and NS, NPS, or BAL was evaluated by calculating the kappa statistic and percent positive agreement. McNemar's exact test was used to compare the proportions of positive results. Results Agreement among TTW and NS, TTW and NPS, and TTW and BAL, was very good for identification of P. multocida, M. haemolytica, and M. bovis. For bovine respiratory syncytial virus (BRSV), agreement with TTW was moderate for NS, good for NPS, and very good for BAL. For bovine coronavirus (BCV), agreement with TTW was moderate for NS and NPS, and good for BAL. McNemar's test was significant only for BCV, indicating that for this pathogen the proportion of positive results from NS and NPS could not be considered comparable to TTW. Conclusions and Clinical Importance This study provides guidance for veterinarians selecting diagnostic tests for antemortem identification of pathogens associated with BRD.

This study describes lower limb musculotendon unit (MTU) forces and powers during sprint cycling. Seven participants (2 female) performed maximal effort cycling at optimal cadence. Three-dimensional motion capture, pedal forces, and surface electromyography (EMG) data were input into participant-calibrated EMG-informed neuromusculoskeletal models to evaluate forces and powers produced by 18 lower-limb MTUs. In the pushing phase (10° − 170°, relative to the right crank at top dead centre), the hip adductor group generated the highest average force (1623.78 ± 980.62 N) and the vastus lateralis generated the highest average positive power (193.11 ± 153.07 W). In the pulling phase (170°-350°), the hip adductor group generated the highest average force (1140.48 ± 482.57 N) and average power (60.47 ± 68.87 W). In the transition phase (350°-10°), the vastus lateralis generated the highest average force (1432.26 ± 1169.40 N) and average power (76.98 ± 71.84 W). MTU power crank cycle-profiles were generated for each participant, highlighting differences in patterns of MTU power generation. Future work is needed to determine if targeted training based on cyclist-specific MTU force and power model outputs would lead to improved sprint cycling performance.

Semi-recumbent cycling performed from a wheelchair is a popular rehabilitation exercise following spinal cord injury (SCI) and is often paired with functional electrical stimulation. However, biomechanical assessment of this cycling modality is lacking, even in unimpaired populations, hindering the development of personalised and safe rehabilitation programs for those with SCI. This study developed a computational pipeline to determine lower limb kinematics, kinetics, and joint contact forces (JCF) in 11 unimpaired participants during voluntary semi-recumbent cycling using a rehabilitation ergometer. Two cadences (40 and 60 revolutions per minute) and three crank powers (15 W, 30 W, and 45 W) were assessed. A rigid body model of a rehabilitation ergometer was combined with a calibrated electromyogram-informed neuromusculoskeletal model to determine JCF at the hip, knee, and ankle. Joint excursions remained consistent across all cadence and powers, but joint moments and JCF differed between 40 and 60 revolutions per minute, with peak JCF force significantly greater at 40 compared to 60 revolutions per minute for all crank powers. Poor correlations were found between mean crank power and peak JCF across all joints. This study provides foundation data and computational methods to enable further evaluation and optimisation of semi-recumbent cycling for application in rehabilitation after SCI and other neurological disorders.

Infectious bronchitis virus (IBV) causes severe economic losses among chicken flocks worldwide. Although IBV molecular surveillance has been conducted in California broilers, seasonal and spatial-temporal trends in IBV prevalence are poorly defined. The goals of this study were to evaluate seasonal and spatial-temporal trends in IBV prevalence and to determine the predominant IBV genotypes obtained over the last 8 yr from a broiler company located in the California Central Valley. In total, 3439 broilers with a suspicion of IBV infection were submitted to the California Animal Health and Food Safety laboratories between January 2012 and February 2020. Swabs from tracheas, kidneys, and cecal tonsils from each submission were independently pooled and screened for IBV using reverse transcriptase quantitative PCR (RT-qPCR). Positive samples were submitted for virus isolation. Viral isolates were subject to a conventional RT-PCR targeting the S1 gene hypervariable region. Positive samples from this RT-PCR were sequenced, and phylogenetic analyses were performed. In total, 1243 pooled swab samples were positive for IBV. Positive results were more frequently detected in fall and winter months compared to spring. Spatial analyses revealed an IBV hot spot in the vicinity of Livingston, and two areas with a low prevalence (i.e., cold spots) around Riverdale. The IBV spatial-temporal distribution identified three significant clusters: one hot spot around Turlock from 2015 to 2016, a second hot spot around Merced from 2012 to 2016, and a cold spot around Fresno from 2017 to 2020. Predominant genotypes changed over time from IBV Cal 99, which was predominant between 2012 and 2014, to IBV 3099 in 2019. Vaccination efforts were initiated in 2018, and as a result, we detected an emerging variant with 92% similarity to CA 3099 in 2020. This work highlights the importance of ongoing surveillance in IBV prevention programs. Surveillance strategies are necessary to monitor trends in diseases such as infectious bronchitis, and the tools used for surveillance need to be sensitive enough to detect new variants and identify spatial-temporal trends.