S oybean vein necrosis virus (Orthotospovirus glycininecrovenae, SVNV) is an ambisense ssRNA virus in the genus Orthotospovirus first identified in Tennessee in 2008 (1). SVNV consists of three segments: S, M, and L. These encode a nucleocapsid protein (N), nonstructural proteins (NSs and NSm), glycoproteins (GN and GC), and an RNA-dependent RNA polymerase (RdRp) (2). The complete sequence of the SVNV17_Auburn_AL isolate was obtained using RNA-Seq and RACE. In 2023, soybean samples exhibiting symptoms of SVNV were collected. Total RNA was extracted from symptomatic leaves using the previous methodology (3), followed by ribosomal RNA depletion using the Illumina Ribo-Zero Plus rRNA Depletion Kit (Illumina, Cat: 20037135). Libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina and sequenced on an Illumina NovaSeq 6000 (150 bp PE, ~47 million reads). Quality control was conducted using FastQC (4), and adapter sequences were removed using BBDuk (https://sourceforge.net/projects/bbmap/). Processed reads were mapped to the SVNV-TN genome (GCA_004789395.1) using Bowtie (5). Variants (depth >80, Phred > 100) were called using BCFtools (6) and FreeBayes tool (7), and the average depth was calculated using SAMtools (6). Identified variants were visualized using IGV v2.3.57 (8). The consensus assembly was generated with BCFtools (6). For all tools, the default parameters were used. Missing terminal nucleotides were filled using RACE: 5′ ends with the Invitrogen 5′ RACE System (ThermoFisher, Cat: 18374058) using segment-specific primers (Table 1), and 3′ ends using E. coli poly(A) polymerase (NEB) followed by SuperScript III (Invitrogen) synthesis. PCR amplification used Phusion (ThermoFisher, F530S), cloned using the CloneJET PCR Cloning Kit, and 12 colonies were Sanger sequenced using an Applied Biosystems 3730xl sequencer. The genome of the SVNV17_Auburn_AL comprises 16,563 bases (2,602 bp [S], 4,948 bp [M], and 9,013 bp [L]) with a GC content of 35% and an average depth of 1,669×. The leaders are 58, 57, and 185 bases, while the trailers are 70, 91, and 30 bases for the S, M, and L segments, respectively. The first six bases (AGAGCA) at the 5′ ends are identical across all three segments and are complementary to the 3′ ends, forming a panhandle similar to other orthotospoviruses (9). Genome comparison between the SVNV17_Auburn_AL isolate and the TN strain revealed 43, 97, and 138 SNPs/indels in the S, M, and L segments (Fig. 1). To determine the impact on the protein level, the ORFs’ sequence was translated with ExPASy (10) and aligned to the TN strain with Clustal

In 2020, an evaluation of the basal level of incidence of tomato spotted wilt (TSW) in peanut was initiated in Alabama. This was done to understand the viral sequence divergence of tomato spotted wilt virus (TSWV) in a heavily managed system. From 2021 to 2023, 172 leaf samples were collected from peanut plants exhibiting symptoms of TSW from Brewton, Fairhope, and Headland, AL, to investigate genetic changes. Additionally, four thrips populations were collected in 2022 and 2023 from Fairhope and Headland, AL. The nucleocapsid protein of TSWV was sequenced from both leaf samples and thrips. A total of 175 nucleocapsids were sequenced, and their amino acid sequences were aligned to identify three conserved mutations compared to TSWV-MT2 (X61799.1) and seven conserved mutations when aligned against TSWV-BR-01 (NC_002051.1). Interestingly, only one conserved mutation was found in the thrips sequences compared to MT2, and no mutations were detected when aligned with BR-01. To identify if mutations caused a phenotype that could be measured, eight nucleocapsids with unique mutations were selected for localization in Nicotiana benthamiana. The localization patterns of these proteins were grouped into three phenotypes based on the observed protein aggregation speed. Small-plot trials assessed TSWV mutations and incidence in relation to planting date, insecticide use, and cultivar. Based on these data, although management strategies are effective at keeping levels of TSWV manageable, the virus can diversify its sequence, which causes changes in the expected phenotype of the nucleocapsid.

Soybean vein necrosis virus (SVNV) is a persistent, propagative, ambisense single-stranded RNA virus in the genus Orthotospovirus, transmitted by Nehydatothrips variabilis. To understand SVNV in the field, 33 samples exhibiting symptoms of SVNV were collected. The N, NSs, and NSm open reading frames (ORFs) were sequenced, revealing amino acid mutations in each gene. The five open reading frames of the SVNV Tennessee strain (N, NSs, NSm, GN, and GC) were fused in frame to GFP for experimentation in both plant and insect cells. N and NSs localize in plants at the cell periphery and nucleus. NSm induces cell death in plant cells, but not in insect cells, where cytoplasmic localization is observed. GN and GC glycoproteins localize to the membranes and display increased cytoplasmic localization in insect cells. The findings of this study contribute to understanding the genes of SVNV and capture sequence changes that have occurred over the past fifteen years.

Tomato spotted wilt virus (TSWV) is a major yield-limiting pathogen of peanut in the southeastern USA. To assess viral variability, field isolates were collected from symptomatic peanut plants at three locations in Alabama. Sequence analysis identified mutations in the non-structural movement protein (NSm) and the nucleocapsid protein (N). Subcellular localization studies showed that NSm fused to GFP (NSm:GFP) localized to plasmodesmata and co-localized with callose deposits by 2days postinfiltration (dpi). By 4 dpi, NSm:GFP formed cytoplasmic aggregates, and callose deposition appeared more consistent with basal plasmodesmatal patterns. The N protein localized to the nuclear periphery and cell margins at 2 dpi and later aggregated in the cytoplasm by 4 dpi. The early callose accumulation associated with NSm expression suggests a potential plant defence response, warranting further investigation.

During the screening of peanuts (Arachis hypogaea L.) for tomato spotted wilt virus (TSWV) in 2024, 36 plant samples expressing symptoms of tomato spotted wilt were collected from three locations in Alabama: Brewton Agricultural Research Unit (BARU), Brewton, AL; Wiregrass Research and Extension Center (WGREC), Headland, AL; and Gulf Coast Research and Extension Center (GCREC), Fairhope, AL. Foliar necrosis symptoms, not typically associated with TSWV, were also observed in all samples. Symptoms varied from moderate light brown to severe dark necrosis along the midveins and lateral veins. Samples were tested for four orthotospoviruses: TSWV, groundnut ringspot virus (GRSV), tomato chlorotic spot virus (TCSV), and soybean vein necrosis virus (SVNV) using ELISA (Agdia Inc., Elkhart, IN). All samples were positive for TSWV and negative for both GRSV and TCSV; however, one sample tested positive for SVNV (BARU 4). SVNV was previously reported not to infect peanuts (Zhou and Tzanetakis 2013). The 36 samples were subjected to RT-PCR using specific primers for the nucleocapsid (N) protein of SVNV (Shehata et al. 2024). Twenty-four of 36 samples were positive for SVNV (12 from BARU, two from WGREC, and 10 from GCREC). The amplified bands showed lower intensity than the positive control, possibly indicating a low titer of SVNV, which may explain the negative ELISAs. Following the manufacturer’sinstructions, these bands were cloned using the CloneJET PCR Cloning Kit (Thermo Fisher Scientific) and sent for Sanger sequencing. The resulting SVNV-N sequences were submitted to GenBank under accessions PQ821900 to PQ821905. The sequences demonstrated 98.19% nucleotide and 96.75% amino acid identities with SVNV from Tennessee (GCF_004789395.1). Nine conserved amino acid mutations were identified compared with the Tennessee strain, resembling those found in soybeans (Shehata et al. 2024). Additionally, RT-PCR was also used for TSWV-N detection (Martin et al. 2025), and 26 samples tested positive (12 from BARU, three from WGREC, and 11 from GCREC), which confirmed the presence of both SVNV and TSWV in positive samples. Further research is necessary to investigate co-infection between SVNV and TSWV, potential genome reassortment, and its mechanisms to understand the interactions between these viruses. These interactions could adversely impact legume production in Alabama, valued at $315 million in 2023 (soybean and peanut, USDA 2023). Since both TSWV and SVNV are transmitted by tobacco thrips (Frankliniella fusca) (Hameed et al. 2022; Keough et al. 2016), this is likely how SVNV was introduced to peanuts. This constitutes the first report on the detection of SVNV in peanuts in the United States, suggesting that SVNV has been adapting to new hosts since its discovery (Tzanetakis et al. 2009; Zhou et al. 2018).

Grapevine leafroll-associated virus-3 (GLRaV-3) is considered the most predominant cause of grapevine leafroll disease (GLD), one of the most destructive viral diseases affecting grapevines and wine production worldwide (Maree et al. 2013). GLRaV-3 is a positive-sense, single-stranded RNA (+ssRNA) virus in the family Closteroviridae, genus Ampelovirus, species tritis (Martelli et al., 2012; Maree et al., 2013). Grape plants (cultivar ‘Errante Noir’) were planted at the Chilton Research and Extension Center (CREC), Chilton County, Alabama, in the spring of 2024. In November 2024, a plant exhibited symptoms of dark-purplish red veins with cupping of the leaf margins, and in July 2025, symptoms of reddish-brown blotches randomly distributed on the foliage appeared (Supplemental Figure 1: A: I-II). To investigate if a pathogen caused this, a sample was sent to Agdia, Inc. (Elkhart, IN, USA), where tissues were subjected to a pathogen screen, including alfalfa mosaic virus (AMV), arabis mosaic virus (ArMV), grapevine fanleaf virus (GFLV), Phytophthora (Phyt), peach rosette mosaic virus (PRMV), strawberry latent ringspot virus (SLRSV), tomato ringspot virus (ToRSV), tobacco ringspot virus (TRSV), GLRaV-3, grapevine pinot gris virus (GPGV), grapevine red blotch-associated virus (GRBaV), and Xylella fastidiosa (Xf). The sample tested negative for all except for GLRaV-3. To confirm GLRaV-3, RT-PCR was used. Total RNA was extracted from 0.1 g petiole tissue from three subsamples using the RNeasy Plant Mini Kit (Qiagen), producing three separate RNA samples. The cDNA was synthesized using SuperScript IV Reverse Transcriptase (Invitrogen), and Platinum Taq DNA Polymerase (Invitrogen) was used to amplify the heat shock protein (HSP70) and the coat protein (CP) using specific primers (Thompson et al. 2019). PCR products were checked on the TapeStation using the D1000 ScreenTape kit (Agilent), and amplicons corresponding to 600 bp (HSP70) and 280 bp (CP) were detected (Supplemental Figure 1: B). Sanger sequencing was conducted at Azenta Life Sciences in both directions. The manufacturer’s instructions were followed in all used kits. Sequences were analyzed using BLASTn (Altschul et al. 1990) to confirm that all three sequences from each gene match GLRaV-3. Sequences were submitted to GenBank (accessions: PX123059-64), and an isolate name was given (GLRaV-3_Chilton-AL.1.1-3). The closest BLAST hit is GLRav3-8415B (KY073324.1) with 100% (HSP70), 98.24% (CP) nucleotide similarities (Supplemental Table 1). The CP sequences of GLRaV-3_Chilton-AL.1.1-3 were compared to 139 isolates of GLRaV-3 from GenBank (Supplemental Table 1) to construct a maximum likelihood tree using methods described by Shehata et al. (2025), with a modification where IQ-TREE was used to construct the tree with 1000 replicates (Nguyen et al. 2015). This tree indicated that GLRaV-3_Chilton-AL.1.1-3 is within group XII, with two isolates from Canada(Supplemental Figure 1: C). Other grape plants planted along with the infected plant at the Chilton Co. site (n=15) were also tested for GLRaV-3 as described above, and all tested negative. This constitutes the first report of GLRaV-3 in Alabama, highlighting the importance of purchasing clean material to prevent introduction of the disease in vineyards, a crop with a total impact of $1.5 billion in Alabama (Good, T. 2023).

This study investigated the effects of coriander seed powder and Lactobacillus acidophilus solution (LAS)
on the growth, nutrient digestibility, blood parameters, and intestinal health of growing rabbits under subtropical
conditions. Forty Californian rabbits, aged 35 days with an average body weight of 588 ± 34 g, were randomly
assigned to four groups: a control group fed a standard diet, a group (T1) receiving the basal diet supplemented with
1.5% coriander seed powder, a group (T2) receiving the basal diet with oral LAS at 1 × 109 CFU/kg, and a group
(T3) receiving both coriander seed powder and LAS. The study measured body weight, daily weight gain, feed
conversion ratio, mortality rate, blood metabolites, nutrient digestibility, and intestinal histomorphology. Results
showed that rabbits fed with 1.5% coriander seed powder had significant improvements in body weight gain, feed
conversion, and a reduction in mortality compared to the control. Both coriander seed powder and LAS improved
blood metabolites, nutrient digestibility, and intestinal health. However, the combination of both additives did not
provide additional benefits over the individual treatments. The findings suggest that either 1.5% coriander seed
powder or LAS can enhance growth performance and health in rabbits under subtropical conditions.

This study examines the prevalence of helminthic infestations, risk factors, and odds ratios in 1 300 cattle,
revealing significant patterns in parasite distribution and influencing factors. Overall, 60.3% of cattle were infested
with one or more types of parasites, with Moniezia spp. being the most prevalent (46.9%), followed by Fasciola
spp. (36.9%), Paramphistomum spp. (26.8%), and Avitellina spp. (10.8%). Among the seasons, winter exhibited the
highest infestation rate (66.1%), and calves under one year of age were more commonly infested (64.2%) than older
cattle (over three years: 51.1%, OR = 0.584 1). Distinct seasonal and age-related patterns were observed for specific
parasites. Fasciola spp. was most prevalent in winter (45.09%) and among the youngest cattle (47.87%), while Paramphistomum
spp. and Moniezia spp. showed less variation across seasons. Avitellina spp. had the lowest infestation
rates in the spring, and cattle older than three years were affected. Co-infections were common, notably between
Fasciola and other parasites, with the highest co-infestation rate observed between Avitellina spp. and Moniezia
spp. Analysis of deworming efficacy indicated higher treatment success for Fasciola spp. and Paramphistomum spp.,
lower odds of response for Avitellina spp., and no significant treatment effect for Moniezia spp. Microscopic and
pathological examinations were also conducted. These results demonstrate the necessity of thorough deworming
procedures along with targeted parasite control to reduce significant health hazards in cattle populations.

This study aimed to evaluate the effect of humic acid (HA) on growth performance, nutrient digestibility, ruminal fermentation, ruminal enzyme activity, and certain blood biochemical parameters. A total of 45 calves (11 ± 0.25 months of age; 280 ± 5.55 kg BW) were divided equally into three groups. The control group received a basal diet composed of a concentrate feed mixture (CFM) with roughage. The treatment groups received the same basal diet supplemented with 1 and 2% humic acid in the CFM, respectively. The study revealed that the treatment groups had significantly lower total dry matter (DM) intake (p = 0.029). The treatment groups showed higher (p = 0.013) ether extract (EE) digestibility and digestible crude protein (DCP) (p = 0.001) than the CFM group. However, ammonia nitrogen (NH3-N, mg/100 mL) was significantly lower (p = 0.012) in the 1% HA and 2% HA groups compared to the CFM group. The 2% HA group had significantly higher (p = 0.001) proportions of acetate and propionate compared to the 1% HA and CFM groups. The treatment groups had significantly higher (p = 0.001) α-amylase, lipase, urease, and protease activities compared to the CFM group. In addition, the concentration of total soluble protein in the rumen liquid was higher (p = 0.006) in the 1% HA group compared to the CFM group. The 2% HA group had higher (p = 0.049) glucose concentrations than the CFM group. In conclusion, humic acid supplementation at 1% or 2% may improve nutrient digestibility and ruminal fermentation efficiency, enhance enzyme activity, and support normal blood profiles in calves.