Days Postinoculation Research Articles

Studies on intra-ocular vaccination of adult cattle with reduced dose Brucella abortus strain-19 vaccine

Brucella abortus vaccines play a central role in bovine brucellosis control with tremendous success worldwide for decades. The study was aimed to evaluate the efficacy of reduced dose (5.0 × 10 9 cfu) of S19 vaccine in adult cattle and its shedding in the milk of vaccinated cattle using molecular techniques. The OIE recommended tests (RBPT, SAT, and iELISA) for brucellosis screening in cattle were used. Seronegative cattle (n = 90) of different age groups (young, old heifers & milking cows, n = 30 each) were selected for the vaccine trials. Antibody titers were recorded at 7th, 21st, 30th, 60th, 90th and 120th days post-vaccination (DPV) to monitor the immune responses following vaccination and at 150th, 180th, 210th and 240th DPB following booster-dose to an intraocular group. The humoral immune responses observed by RBPT and ELISA, proved that antibody titers persisted in s/c group compared to the i/o group in all categories. The IFN-γ stimulation (CMI) due to reduced dose vaccination was noticed early as 30th in all groups and declined after 90th DPV, with higher IFN-γ stimulation among the s/c group. The Bcsp31 and IS711 targeted PCR detected the presence of Brucella DNA in milk samples (n = 120) from the vaccinated cows (n = 30) and confirmed by qPCR (TaqMan assay) at 30th, 60th, 90th and 120th DPV. A Significant number, 70% (7/10) was detected in s/c by qPCR. BCSP31 sequence was deposited at NCBI GenBank (accession no. MK881173-6). PCR and qPCR techniques could provide a reliable diagnosis of brucellosis from milk. The intraocular route remains the safer route for vaccinating adult cattle than subcutaneous.

Genetic and pathogenic characteristics of a novel infectious bronchitis virus strain in genogroup VI (CK/CH/FJ/202005)

Infectious bronchitis virus (IBV) has a significant impact on the poultry industry, and genogroup VI (GVI) IBVs, such as the TC07−2 strain, were reported in China since 2007. We isolated a novel strain, CK/CH/FJ/202005 (henceforth 202005), from broilers that were vaccinated with different attenuated IBV strains (H120, 4/91, or QX) in Fujian, China during 2020. Based on comparison with the SNU8065 strain (GI-22), the 1ab genes were the locations of recombination in this new isolate. Pathogenicity testing of 1-day-old and 15-day-old specific pathogen-free (SPF) chickens indicated varying severities of air sacculitis beginning 5 days-post-inoculation (DPI). Histopathological analysis indicated that tracheal lesions started at 5 DPI and persisted throughout the 30-day experiments in 1-day-old infected chickens. Virus re-isolation and viral load tests indicated this strain was mainly in the trachea, and not the kidneys. Our findings showed that the 202005 strain was pathogenic in 1-day-old and 15-day-old chickens. These results should be considered when developing strategies to control IBV infections.

First Report of Sclerotinia Fruit and Crown Rot Caused by Sclerotinia sclerotiorum on Short-Day Strawberry in Maryland.

With over 1,000 producers in the Mid-Atlantic region, short-day strawberry (Fragaria x Ananassa Duch.) represents an economically important industry. In spring of 2016, a white rot was observed on strawberry stem, crown, and fruit tissue at two Maryland farms. Light to dark brown necrotic lesions, white mycelium and medium to large black sclerotia were present on petioles and fruit (Fig. 1). These symptoms were most consistent with Sclerotinia fruit rot and crown rot caused by Sclerotinia sclerotiorum (Lib.) de Bary (Maas 1998), not previously described in the region (Farr and Rossman 2019). Lesion margins were excised, surface disinfested (70% EtOH for 30 s and 0.1% NaClO for 1 min) and placed onto potato dextrose agar (PDA) amended with 0.03% tetracycline. Single hyphal tip cultures were obtained which formed fluffy white to tan mycelium, no asexual conidia, and numerous pigmented sclerotia resembling S. sclerotiorum (Bolton 2006). DNA was extracted from five isolates and the internal transcribed spacer (ITS) region was amplified by PCR using primers ITS1/ITS4; all five ITS sequences (533 bp) had 99-100% similarity with S. sclerotiorum (GQ375746) in Genbank. Sequences of SL1380 and SL1382 were submitted to NCBI under the accession numbers MK429976 and MK429977, respectively. These two isolates were evaluated for abilities to cause fruit and stem petiole rot. Pathogenicity on fruit was examined by placing a 5 mm plug from five-day-old cultures on surface disinfested fruits, with mock-inoculated (PDA plug only) and non-inoculated controls. Fruit were placed in incubators (~98% RH, 22°C, 10:14 hrs L:D) and arranged in a complete block design with three blocks and five fruit/treatment/block; the study was conducted twice. White hyphae and sclerotia formed on 41 to 53% of fruit inoculated with both isolates, necrotizing fruit within 6 days post inoculation (DPI); Sclerotinia fruit rot did not develop in controls. Pathogenicity on petioles was examined in a replicated study on three-month-old cv. Flavorfest using a petiole inoculation method described in (Peltier and Grau 2008); mock-inoculated stems were placed in sterile PDA. Plants were arranged in the greenhouse (~20-28°C; 12:12 hrs L:D) in a randomized complete block design with three blocks and three plants/treatment. Dark brown necrosis averaging 5.8 ± 0.2 mm in length developed on 100% of plants in both isolate treatments within 3 DPI and by 6 DPI had expanded to 42.1 ± 4.8 mm on average, spreading into the crown on some plants. Necrosis did not extend beyond 2 mm from the cut tip in controls. The pathogen was recovered from surface disinfested tissue for both isolates. In a subsequent study, stem rot susceptibility was compared among four short-day strawberry cultivars common to the region, cvs. Flavorfest, Chandler, Allstar and Honeoye (same method as above). Stem lesions were 2 to 3 times longer on cv. Flavorfest (41.78(ave) ± 6.73(se) mm) compared to all other cultivars (13.68(ave) ± 1.56(se) mm) (P < 0.0001, Tukey's test). To our knowledge, this is the first report of both Sclerotinia fruit and crown rot on strawberries anywhere in the mid-Atlantic region. Our results indicate that cv. Flavorfest, a recently developed cultivar with expanding acreage, may be more susceptible to Sclerotinia. These are both unusual diseases circumglobally, especially Sclerotinia fruit rot (Farr and Rosman 2019, Maas 1998); the new occurrence in this region may be associated with increased use of this cultivar.

Histopathology of the Plasmodiophora brassicae-Chinese Cabbage Interaction in Hosts Carrying Different Sources of Resistance.

Clubroot is a serious soil-borne disease of crucifers caused by the obligate parasite Plasmodiophora brassicae. The genetic basis and histopathology of clubroot resistance in two Chinese cabbage (Brassica rapa ssp. pekinensis) inbred lines Bap055 and Bap246, challenged with pathotype 4 of P. brassicae, was evaluated. The Chinese cabbage cultivar “Juxin” served as a susceptible check. The resistance in Bap055 was found to be controlled by the CRa gene, while resistance in Bap246 fit a model of control by unknown recessive gene. Infection of the roots by P. brassicae was examined by inverted microscopy. Despite their resistance, primary and secondary infection were observed to occur in Bap055 and Bap246. Primary infection was detected at 2 days post-inoculation (DPI) in “Juxin,” at 4 DPI in Bap055, and at 6 DPI in Bap246. Infection occurred most quickly on “Juxin,” with 60% of the root hairs infected at 10 DPI, followed by Bap055 (31% of the root hairs infected at 12 DPI) and Bap246 (20% of the root hairs infected at 14 DPI). Secondary infection of “Juxin” was first observed at 8 DPI, while in Bap055 and Bap246, secondary infection was first observed at 10 DPI. At 14 DPI, the percentage of cortical infection in “Juxin,” Bap055 and Bap246 was 93.3, 20.0, and 11.1%, respectively. Although cortical infection was more widespread in Bap055 than in Bap246, secondary infection in both of these hosts was restricted relative to the susceptible check, and the vascular system remained intact. A large number of binucleate secondary plasmodia were observed in “Juxin” and the vascular system was disrupted at 16 DPI; in Bap055 and Bap246, only a few secondary plasmodia were visible, with no binucleate secondary plasmodia. The defense mechanisms and expression of resistance appears to differ between Chinese cabbage cultivars carrying different sources of resistance.

Two bicistronic DNA vaccines against Vibrio anguillarum and the immune effects on flounder Paralichthys olivaceus.

Chemokines are cytokines that can promote the activation and migration of immune cells, and increase the recognition of antigen by antigen-presenting cells (APC). Previous studies showed that a DNA vaccine can induce humoral and cellular immune responses of flounder after immunization. To explore the improvement of chemokines on the efficiency of OmpK vaccine, two bicistronic DNA candidate vaccines were constructed and the immune responses they induced in the flounder were investigated by reverse transcription polymerase chain reaction (RT-PCR), indirect immunofluorescent assay (IFA), H&E staining, flow cytometry (FCM), and quantificational real-time polymerase chain reaction (qRT-PCR). pBudCE4.1 plasmid as an expression vector, bicistronic DNA vaccines encoding OmpK gene and CC-motif ligand 4 gene (p-OmpK-CCL4), or Ompk gene and CC-motif ligand 19 gene (p-OmpK-CCL19) were successfully constructed. The results showed that two bicistronic DNA vaccines expressed Ompk protein of Vibrio anguillarum and CCL4/CCL19 proteins of flounder both in vitro and in vivo. After immunization, a large number of leucocytes in muscle were recruited at the injection site in treatment groups. The constructed vaccines induced significant increases in CD4-1+ and CD4-2+ T lymphocytes, and sIgM+ B lymphocytes in peripheral blood, spleen, and head kidney. The percentage of T lymphocytes peaked on the 14th post-vaccination day whereas that of B lymphocytes peaked in the 6th post-vaccination week. Moreover, the expression profiles of 10 immune-related genes increased in muscles around the injection site, spleen, and head kidney. After the challenge, p-OmpK-CCL4 and p-OmpK-CCL19 conferred a relative percentage survival (RPS) of 74.1% and 63.3%, respectively, higher than p-OmpK alone (40.8%). In conclusion, both CCL4 and CCL19 can improve the protection of p-OmpK via evoking local immune response and then humoral and cellular immunity. CCL4 and CCL19 will be potential molecular adjuvants for use in DNA vaccines.Electronic supplementary materialSupplementary material (Supplementary Material) is available in the online version of this article at 10.1007/s00343-021-1092-z.

Purification of Tomato Bushy Stunt Virus Particles by One-Step Hydroxyapatite Column Chromatography

The main aim of this work was to develop a time-saving and cost-effective purification method of infectious plant viral nanoparticles. Virions of Tomato bushy stunt virus (TBSV), which is a member of Tombusvirus genus, were purified by one-step Bio-gel HT Hydroxyapatite (HA) column chromatography. Extracts from Nicotiana benthamiana plants infected with TBSV were directly loaded onto the HA column and eluted by 10 mM sodium phosphate buffer (pH 6.8). A specificity of virions has been confirmed by immunoblotting and electron microscopy. Homogeneity of virions was tested by SDS-PAGE, where only 41 kDa polypeptide bands referring to the capsid protein of TBSV were detected by Coomassie staining. The biological infectious activity of a purified material was demonstrated by observing TBSV-specific symptoms observed in N. benthamiana plants at 7‒10 days of post-inoculation (dpi). Moreover, purified virions were used for immunization of the BALb/c mouse to raise primary antibodies against the TBSV virus. Our results show that in low concentrations of sodium phosphate buffer total proteins extracted from infected plants adsorb to HA sorbent, while viral particles do not adsorb to the HA matrix and flow throw column due to Ca2+ ions implicated in TBSV virions’ structure. This highly effective and simple virus purification protocol can also be used for the isolation of other plant virions.

The Fungus Metarhizium sp. BCC 4849 Is an Effective and Safe Mycoinsecticide for the Management of Spider Mites and Other Insect Pests.

Simple SummaryThe spider mite is a destructive pest of various crops during warm and dry conditions in tropical countries, including Thailand. The pest is difficult to control despite synthetic acaricide use. In the field, insect populations gradually develop resistance to synthetic pesticides over long-term use. The use of entomopathogenic fungi is more human- and environmentally friendly. We searched and identified a potential fungal isolate from a culture collection, focusing on the genus Metarhizium. Metarhizium sp. BCC 4849 not only controls the spider mite but also plays a significant role as a natural regulator of other insect pests. Here, we investigated its infection process on the mite, optimized a conidial formulation for extended shelf life, and conducted toxicological assays in animals to assess its biosafety in humans. The fungal genome has been sequenced. The genomic data indicated that oxidoreduction proteins; zinc-, heme-, and iron-binding proteins; and transmembrane transporters are abundant in the genome.Five isolates of Metarhizium sp. were evaluated for their pathogenicity against the spider mite (Tetranychus truncatus Ehara) (Acari: Tetranychidae) and Metarhizium sp. BCC 4849 resulted in the highest mortality (82%) on the 5th day post-inoculation (DPI). Subsequent insect bioassay data indicated similar high virulence against five other insects: African red mites (Eutetranychus africanus Tucker) (Acari: Tetranychidae), bean aphid (Aphis craccivora Koch) (Hemiptera: Aphididae), cassava mealybug (Phenacoccus manihoti Matile-Ferrero) (Hemiptera: Pseudococcidae), sweet potato weevil (Cylas formicarius Fabricius) (Coleoptera: Brentidae), and oriental fruit fly (Bactrocera dorsalis Hendel) (Diptera: Tephritidae), at mortalities of 92–99%, on 3rd–6th DPI, and in laboratory conditions. The pathogenicity assay against E. africanus in hemp plants under greenhouse conditions indicated 85–100% insect mortality on 10th DPI using the fungus alone or in combination with synthetic acaricide. Genome sequencing of Metarhizium sp. BCC 4849 revealed the high abundance of proteins associated with zinc-, heme-, and iron-binding; oxidation-reduction; and transmembrane transport, implicating its versatile mode of interaction with the environment and adaptation to various ion homeostasis. The light and scanning electron microscopy indicated that at 24 h post inoculation (PI), adhesion and appressorial formation occurred, notably near the setae. Most infected mites had stopped moving and started dying by 48–72 h PI. Elongated hyphal bodies and oval blastospores were detected in the legs. At 96–120 h PI or longer, dense mycelia and conidial mass had colonized the interior and exterior of dead mites, primarily at the bottom than the upper part. The shelf-life study also indicated that conidial formulation combined with an oxygen-moisture absorber markedly enhanced the viability and germination after storage at 35 °C for four months. The fungus was tested as safe for humans and animals, according to our toxicological assays.

Pulmonary lesions induced by SARS-CoV-2 infection in domestic cats.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of coronavirus disease 2019, which ranges from fatal disease in some to mild or subclinical in most affected individuals. Many recovered human patients report persistent respiratory signs; however, lung disease in post-acute infection is poorly understood. Our objective was to describe histologic lung lesions and viral loads following experimental SARS-CoV-2 infection in 11 cats. Microscopic evaluation at 3, 6, 10, or 28 days postinoculation (DPI) identified mild to moderate patchy interstitial pneumonia, bronchiolar epithelial damage, and occlusive histiocytic bronchiolitis. Based on immunohistochemistry, alveolar septal thickening was due to CD204-positive macrophages, fewer B and T lymphocytes, type II pneumocytes, and capillary proliferation with a relative dearth of fibrosis. In blood vessel endothelium, there was reactive hypertrophy or vacuolar degeneration and increased MHC II expression at all time points. Unexpectedly, one cat from the 28 DPI group had severe subacute regionally extensive lymphohistiocytic pneumonia with multifocal consolidation, vasculitis, and alveolar fibrin. Reverse transcriptase-quantitative polymerase chain reaction identified SARS-CoV-2 RNA within the lung at 3 and 6 DPI, and viral RNA was below the limit of detection at 10 and 28 DPI, suggesting that pulmonary lesions persist beyond detection of viral RNA. These findings clarify our comparative understanding of disease induced by SARS-CoV-2 and suggest that cats can serve as an informative model to study post-acute pulmonary sequelae.

Pathogenicity, infective dose and altered gut microbiota in piglets infected with porcine deltacoronavirus

Porcine deltacoronavirus (PDCoV) is an emerging swine enteropathogenic coronavirus that cause severe diarrhea, resulting in high mortality in neonatal piglets. Little is known regarding the pathogenicity of PDCoV in different infective dose and the dynamic changes in the composition of the gut microbiota in PDCoV-induced diarrhea piglets. In this study, 5-day-old piglets were experimentally infected with different dose of PDCoV. The challenged piglets developed typical symptoms, characterized by acute and severe watery diarrhea from 1 to 8 days post-inoculation (DPI), and viral shedding was detected in rectal swab until 11 DPI. Tissues of small intestines displayed significant macroscopic and microscopic lesions with clear viral antigen expression. However, no significant differences among groups were found in challenged piglets. Then alteration in gut microbiota in the jejunum and colon of PDCoV infected-piglets were analyzed using 16S rRNA sequencing. PDCoV infection reduced bacterial diversity and richness, and significantly altered the structure and abundance of the microbiota from the phylum to genus. Fusobacterium, and Proteobacteria was significantly increased (P < 0.05), while the abundance of Bacteroidota was markedly decreased in the infected-piglets. Furthermore, microbial function prediction indicated that the changes in intestinal bacterial also affected the immune system, excretory system, circulatory system, neurodegenerative disease, cardiovascular disease, xenobiotics biodegradation and metabolism, etc. These findings suggest that regulating gut microbiota community may be an effective approach for preventing PDCoV infection.

Cucumber powdery mildew detection using hyperspectral data

This study aimed to understand the spectral changes induced by Podosphaera xanthii, the causal agent of powdery mildew, in cucumber leaves from the moment of inoculation until visible symptoms are apparent. A principal component analysis (PCA) was applied to the spectra to assess the spectral separability between healthy and infected leaves. A spectral ratio between infected and healthy leaf spectra was used to determine the best wavelengths for detecting the disease. Additionally, the spectra were used to compute two spectral variables [i.e., the red-well point (RWP) and the red-edge inflexion point (REP)]. A linear support vector machine (SVM) classifier was applied to certain spectral features to assess how well these features can separate the infected leaves from the healthy ones. The PCA showed that a good separability could be achieved from 4 days post-inoculation (DPI). The best model to fit the RWP and REP wavelengths corresponded to a linear model. The linear model had a higher adjusted R2 for the infected leaves than for the healthy leaves. The SVM trained with five first principal components scores achieved an overall accuracy of 95% at 4 DPI (i.e., two days before the visible symptoms). With the RWP and REP parameters, the SVM accuracy increased as a function of the day of inoculation, reaching 89% and 86%, respectively, when symptoms were visible at 6 DPI. Further research must consider a higher number of samples and more temporal repetitions of the experiment.

Comparative susceptibility of eastern cottontails and New Zealand white rabbits to classical rabbit haemorrhagic disease virus (RHDV) and RHDV2.

Rabbit haemorrhagic disease virus (RHDV) is associated with high morbidity and mortality in the European rabbit (Oryctolagus cuniculus). In 2010, a genetically distinct RHDV named RHDV2 emerged in Europe and spread to many other regions, including North America in 2016. Prior to this study it was unknown if eastern cottontails (ECT(s); Sylvilagus floridanus), one of the most common wild lagomorphs in the United States, were susceptible to RHDV2. In this study, 10 wild-caught ECTs and 10 New Zealand white rabbits (NZWR(s); O. cuniculus) were each inoculated orally with either RHDV (RHDVa/GI.1a; n=5 per species) or RHDV2 (a recombinant GI.1bP-GI.2; n=5 per species) and monitored for the development of disease. Three of the five ECTs that were infected with RHDV2 developed disease consistent with RHD and died at 4 and 6 days post-inoculation (DPI). The RHDV major capsid protein/antigen (VP60) was detected in the livers of three ECTs infected with RHDV2, but none was detected in the ECTs infected with RHDV. Additionally, RHD viral RNA was detected in the liver, spleen, intestine and blood of ECTs infected with RHDV2, but not in the ECTs infected with RHDV. RHD viral RNA was detected in urine, oral swabs and rectal swabs in at least two of five ECTs infected with RHDV2. One ECT inoculated with RHDV2 seroconverted and developed a high antibody titre by the end of the experimental period (21 DPI). ECTs inoculated with the classic RHDV did not seroconvert. In comparison, NZWRs inoculated with RHDV2 exhibited high mortality (five of five) at 2 DPI and four of five NZWRs inoculated with RHDV either died or were euthanized at 2 DPI indicating both of these viruses were highly pathogenic to this species. This experiment indicates that ECTs are susceptible to RHDV2 and can shed viral RNA, thereby suggesting this species could be involved in the epidemiology of this virus.

Thyroid hormone suppression in feeder pigs following polymicrobial or porcine reproductive and respiratory syndrome virus-2 challenge.

Thyroid hormones are powerful regulators of growth, development, and basal metabolic rate and can be dysregulated under conditions of severe stress or illness. To understand the role of these hormones in porcine disease response, serum samples were obtained from three batches of nursery-aged pigs (n = 208) exposed to a natural polymicrobial disease challenge with an array of bacterial and viral pathogens. Levels of total thyroxin (T4) and triiodothyronine (T3) assessed in sera by radioimmunoassay, decreased significantly by 14 days post-exposure (DPE). Levels of T3 partially rebounded by 48 DPE, while T4 levels remain depressed. Post-exposure T3 and T4 levels were positively correlated with acute and long-term average daily gain (ADG). Cross-sectional sampling of animals maintained at the high health source farms, showed no equivalent change in either hormone when managed under standard industrial conditions. To further elucidate the effect of porcine reproductive and respiratory syndrome virus (PRRSV)-infection on thyroid hormone levels, archived sera over 42 days post inoculation (DPI) from nursery pigs (N = 190) challenged with one of two PRRSV2 strains by the PRRS Host Genetics Consortium were similarly assessed, with animals selected in a two-by-two design, to investigate biological extremes in ADG and viral load (VL). All animals showed a similar decrease in both thyroid hormones reaching a minimum at 7 DPI and returning to near pre-challenge levels by 42 DPI. Post-challenge T3 and T4 levels were significantly greater in high ADG groups, with no significant association with VL or strain. The results of this study demonstrate porcine susceptibility to thyroid disruption in response to disease challenge and demonstrate a relationship between this response and growth performance.

COVID-19 vaccination and a severe pain in the neck

A 40-year-old woman developed malaise and severe anterior neck pain 12 hours after the second dose of COVID-19 mRNA vaccine (Pfizer/BioNTech). The pain peaked and plateaued at the fourth post-vaccination day. Her GP prescribed ibuprofen and azithromycin for presumed bacterial pharyngitis, with no avail. She visited our endocrine clinic 26 days post-vaccination with ongoing symptoms including bony aches, exhaustion, emotional lability, palpitations and hyperhidrosis. Upon general examination she was haemodynamically stable but with significant neck tenderness (visual acuity score = 8/10).

Experimental porcine astrovirus type 3-associated polioencephalomyelitis in swine.

Porcine astrovirus type 3 (PoAstV3) is an emerging virus in the family Astroviridae that has been recently associated with polioencephalomyelitis/encephalitis. Herein, we describe the experimental oral and intravenous inoculation of an infectious central nervous system (CNS) tissue homogenate containing PoAstV3 to cesarean-derived, colostrum-deprived pigs, and the subsequent development of clinical signs, histologic lesions, specific humoral immune response, and detection of viral particles by electron microscopy (EM) and viral RNA by RT-qPCR (reverse transcriptase quantitative polymerase chain reaction) and in situ hybridization (ISH). IgG against a portion of the PoAstV3 ORF2 capsid was first detected at 7 days post-inoculation (DPI) in 2 of 4 inoculated animals and in all inoculated animals by 14 DPI. At 21 and 28 DPI, 2 of 4 inoculated animals developed ataxia, tetraparesis, and/or lateral recumbency. All inoculated animals had histologic lesions in the CNS including perivascular lymphoplasmacytic cuffs, multifocal areas of gliosis with neuronal necrosis, satellitosis, and radiculoneuritis, and PoAstV3 RNA as detected by RT-qPCR within multiple anatomic regions of the CNS. Consistent viral structures were within the soma of a spinal cord neuron in the single pig examined by EM. Of note, PoAstV3 was not only detected by ISH in neurons of the cerebrum and spinal cord but also neurons of the dorsal root ganglion and nerve roots consistent with viral dissemination via axonal transport. This is the first study reproducing CNS disease with a porcine astrovirus strain consistent with natural infection, suggesting that pigs may serve as an animal model to study the pathogenesis of neurotropic astroviruses.

Efficacy of a Synergistic Blend of Organic Acids and ß-1,4 Mannobiose on Cecal Salmonella Counts and Growth Performance in Salmonella Challenged Broiler Chickens: A Meta-Analysis.

Simple SummarySalmonella may cause serious diarrhea in consumers of poultry products. Control of Salmonella in poultry with antibiotics has caused antimicrobial resistance issues. Salmonella can colonize the gut of chickens, meaning that an in-feed solution may prevent infection. A synergistic blend containing organic acids and ß-1,4 mannobiose as main ingredients were developed to reach different parts of the gut. It was hypothesized that this synergistic blend decreases Salmonella contamination of chickens. Several, non-published, studies have been performed to test the effect of this synergistic blend in chickens infected with Salmonella. The results of these studies were combined into a large meta-analysis to draw conclusions regardless of study design and geographical location. This state-of-the-art statistical method did show that feeding the synergistic blend to chickens could decrease Salmonella in comparison to a control diet. This decrease was most clear during the first 14 days after initiation of the Salmonella infection. In addition, the birds did grow more efficiently when the synergistic blend was fed.This study aimed at investigating the effect of a functional synergistic feed additive blend, containing organic acids and ß-1,4 mannobiose, on cecal Salmonella counts and growth performance of broiler chickens. A meta-analysis combining 13 individual studies, executed in Salmonella-challenged broilers comparing a control diet with and without the synergistic blend, was performed. Cecal Salmonella colonies and overall growth performance were measured. Raw data from all studies were combined and analyzed using PROC MIXED in SAS, taking the within and between-study variation into account. In the first 14 days post-inoculation (DPI), cecal Salmonella was reduced by 0.429 log CFU/g (p = 0.011, n = 10 comparisons from five studies) when feeding the synergistic blend compared to the control group. During 15–34 DPI, the overall means were not different between treatments (0.069 log CFU/g; p = 0.519, n = 12 comparisons from eight studies). The feed conversion ratio was improved when feeding the synergistic blend compared to the control diet (1.474 vs. 1.482, respectively; p = 0.002). In conclusion, feeding a synergistic blend, containing organic acids and ß-1,4 mannobiose amongst other ingredients, reduced cecal Salmonella counts during the first 14 DPI and improved growth performance of Salmonella challenged broilers compared to a control diet.

Association analysis in a germplasm panel of watermelon (Citrullus lanatus (Thunb.) Matsum and Nakai) predominantly from India confirms a QTL conferring resistance to Fusarium oxysporum f. sp. niveum race 1

ABSTRACT An experiment was conducted to screen a germplasm panel comprising 331 Citrullus sp. accessions against Fon race 1 for identification of resistance and to validate a candidate QTL (Fon-1) region conferring resistance. The accessions were screened through artificial inoculation of spore suspension during the rainy season of 2017. A genotype with ≥67% survival at 28 days post-inoculation (DPI) was categorised as resistant. A total of 41 accessions were identified to possess resistance to Fon race 1. Of these, nine accessions were highly resistant (mean survival ≥90% at 28 DPI). The reactions of these accessions were also confirmed in another bioassay. These accessions need deployment in future watermelon breeding programmes. Six markers, located within the previously reported Fon-1 locus, were tested for their association with resistance and among them, InDel-4, located in the 5ʹ promoter region of an ABC Transporter protein, was found to be most significantly associated and explained maximum phenotypic variation, proving its relevance for marker-assisted introgression of resistance to Fon race 1 into elite commercial backgrounds of watermelon.

A Methodology to Quantify Resilience in Growing Pigs.

Simple SummaryThe concept of resilience can be defined as the ability of animals to maintain their productivity despite the stressors that might occur during the rearing period. Despite the growing interest in the genetic determinism of resilience and its possible inclusion in selection objectives, there is no straightforward method to measure this trait. Several quantifying methods estimate resilience based on productivity such as body weight or milk production, or non-productivity related traits such as immunity or stress responses. The objective of this study was to elaborate novel resilience indicators in growing pigs based on both productivity (body weight) and non-productivity (acute-phase proteins) related traits. We propose the deviation from the expected growth curve and the increment of the acute-phase protein haptoglobin, after applying a common vaccine, as resilience indicators in growing pigs under standard rearing conditions. We showed that the suggested resilience indicators are under a genetic control, show a substantial variability in the population, and may be improved through selection.There is a growing concern about the genetic determinism of resilience and its possible implementation in breeding programs. The objective of our study was to elaborate novel resilience indicators in growing pigs based on the deviation from the expected growth curve and the increment of the acute-phase protein haptoglobin (HP) after applying a common vaccine. A total of 445 pigs were vaccinated with an attenuated Aujeszky vaccine at 12 weeks of age. Deviation from the expected body weight (ΔBW) given the growth curve of unvaccinated pigs at 28 days post-vaccination (DPV) and the increment of HP at 4 DPV (ΔHP) were suggested as resilience indicators. Challenged pigs that maintained their productivity and had a minor activation of HP were deemed resilient, whereas pigs that had low ∆BW values and a high activation of HP were deemed susceptible. Pigs were also classified based on ∆BW and ∆HP relative to the expected BW at 28 DPV and to the basal level of HP, respectively. The concordance was high between both methods, indicating that ΔBW and ΔHP are not sensitive to the animal’s expected BW nor the basal level of HP. The heritability estimates were moderate for ∆BW (0.33) and low-to-moderate for ∆HP (0.16). Our study suggests ΔBW and ΔHP as novel resilience indicators in pigs. The suggested indicators capture different aspects of resilience, are easy to measure, and are genetically controlled. Thus, they may be improved through selective breeding. Further analyses are needed to validate our findings.

Experimental West Nile Virus Infection in Northern Bobwhite Quail (Colinus virginianus).

West Nile virus (WNV) has been implicated in regional declines of numerous North American bird species, although its potential impact upon many species, including some game birds, remains unknown. Specifically, information about susceptibility to infection and infection outcome are crucial to assessing health risks. Northern bobwhite quail (Colinus virginianus) are a popular and common game bird across much of the United States, as well as in captive breeding programs and as backyard birds. Two age groups of bobwhites were subcutaneously inoculated with WNV and euthanatized on 15 days postinoculation (DPI). Three of 10 inoculated 5-wk-old and 4/10 inoculated 15-wk-old birds developed detectable viremia titers during 1-5 DPI, with low peak titers (101.7-103.0 plaque-forming units [PFU]/ml). Three of 10 inoculated 5-wk-old and 1/10 inoculated 15-wk-old birds shed low viral titers (peak 100.7-101.8 PFU/swab) either orally or cloacally or both for limited periods from 2 to 6 DPI. All inoculated birds (n = 20) remained apparently healthy and seroconverted by 15 DPI. No infectious virus was detected in select tissues: heart, kidney, brain, skeletal muscle, spleen (15-wk-old group only), and feathers from any of the bobwhites. No sham-inoculated, contact control birds (n = 8) became viremic or had virus isolated from tissues or swabs. The most consistent microscopic lesion was minimal to mild, lymphoplasmacytic myocarditis (6/10 in 5-wk-olds; 5/10 in 15-wk-olds). Immunohistochemical labeling was most often in macrophages in spleen and bone marrow, likely reflective of clearance of infection. There were no statistically significant differences in the peak viremia and shedding titers between age groups and no differences in the development of WNV-associated lesions between the two age groups. These results suggest that WNV is unlikely to pose a health risk to bobwhites and that bobwhites likely are an incompetent reservoir host species in WNV transmission.

Preliminary Findings of New Citrus Rootstocks Potentially Tolerant to Foot Rot Caused by Phytophthora

Phytophthora spp. are one the most common soil-borne pathogens in citrus crops, in which Phytophthoranicotianae and P. citrophthora are the most relevant species, causing disease problems worldwide, such as foot rot and gummosis of the trunk, branch canker, brown rot of fruit, feeder root rot in orchards, and seedling damping-off in nurseries. Phytophthora-tolerant citrus rootstocks are essential for its control and for the success of the citrus industry. The aim of this study was to determine the susceptibility of new citrus rootstocks with low HLB incidence to Phytophthora diseases. Thus, plants of several commercial and new citrus rootstocks originating in different breeding programs were inoculated with an isolate of P. nicotianae. Thirty days post inoculation (DPI), the damage of lesion length in stem was measured for each plant. These results displayed a different susceptibility response to the damage caused by P. nicotianae among the citrus rootstocks tested. Thus, eleven new citrus rootstocks (B11R3T25, B11R5T25, B11R5T49, B11R5T60, B11R5T64, N40R1T18, N40R1T19, N40R3T25, WGFT + 50-7, UFR-6, and CL-5146), which have not been previously studied against Phytophthora diseases, improved the tolerance effect of Carrizo citrange. Our findings provide useful information for citrus growers on rootstock selection to address incidence problems caused by Phytophthora spp.

PSIV-7 Effect of maternal infection and dietary supplementation of isoflavones on offspring postnatal immune function

Abstract The objective was to determine the effect of maternal infection and dietary supplementation of isoflavones during gestation on offspring immune function. First parity gilts (n = 24) at gestational day (GD) 65 were allotted to one of three treatments: uninfected and fed a diet devoid of isoflavones (CON), infected with porcine reproductive and respiratory syndrome virus (PRRSV) and fed the control diet (POS), or infected with PRRSV and fed a diet supplemented with 1,500 ppm soy isoflavones (ISF). Gilts were inoculated intranasally with saline or 2.5×104 TCID50/mL of suspended live PRRSV (NADC20 strain) on GD 70. At farrowing, 4 offspring pigs weighing closest to the litter average were selected for postnatal immune challenges. Innate immunity was determined by serum TNF-α concentrations after lipopolysaccharide (LPS) injection (5 µg/kg BW) on postnatal d (PND) 52. Adaptive immunity was determined in the same pigs by peripheral T cell population shifts following vaccination against porcine circovirus-2 on PND 59. Whole blood was collected on days post-vaccination (DPV) 0, 7, and 14 for T cell phenotyping. In the innate immune challenge, all groups experienced increased TNF-α after LPS injection with peak response at 4 h, with ISF eliciting higher (P = 0.04) TNF-α concentrations than CON, and POS being intermediate. Also, ISF had higher (P &amp;lt; 0.01) TNF-α concentrations at both 0- and 8-h post-challenge compared with CON and POS. Prior to vaccination, ISF had elevated memory T cells and reduced total and naïve T cells (P ≤ 0.02). At DPV 14, the reduced total and naïve T cells persisted in ISF pigs. Overall, these data indicate that maternal PRRSV infection alone does not predispose pigs to an exaggerated immune response. However, maternal isoflavone supplementation during PRRSV infection altered offspring innate and adaptive immune responses.