The present study was conducted to characterize the infectious bursal disease virus (IBDV) circulating in clinically diseased broiler chicken flocks with previous vaccination history during 2015-2016 in Egypt. IBDVs were isolated from 48 out of 63 of the investigated bursae from 10 flocks onto embryonated chicken eggs (ECEs) and verified by reverse transcriptase-poly- merase chain reaction (RT-PCR). Histopathologically, bursae lesions revealed some lymphocytes depletion as well as the presence of vesicles in the lining epithelium. The hyper variable region (HVR) of VP2 and VP1 genes of the 10 isolates (1 isolate/flock) were partially sequenced and subjected to comparative alignment and phyologenetic analysis. Phylogenetically, IBDV isolates were clustered into two distinct genetic lineages: variants of classical virulent (cv) and very viru- lent (vv) IBDV strains based on VP1 and VP2 amino acid (aa) sequences. Alignment analysis of HVR-VP2 aa sequences has demonstrated that the vvIBDV isolates have the conserved residues of the vvIBDV pathotype (A222, I242, and I256), while, the cvIBDV isolates have the same aa sequences of the classical attenuated vaccine strain (D78). Expected single point mutation occurred at position 253 (H253N). All previously characterized isolates were re-subjected to molecular analysis with VP1 protein due to its correlation with virulence and pathogenicity of IBDVs. vvIBDV isolates have the conserved tripeptide (TDN), while, the cvIBDV isolates have aa substitutions at conserved tripeptide including NEG at 145-147 amino acid. The present study has demonstrated that variants of classical virulent and very virulent IBDV circulated among vaccinated flocks in Egypt during 2015-2016.

The full-length cDNA of LeTIR1 gene was isolated from tomato with EST-based in silico cloning followed by RACE amplification. LeTIR1 contained an open reading frame (ORF) 1872 bp long, encoding 624 amino acid residues. The predicted protein LeTIR1 had one F-box motif and eleven leucine-rich repeats (LRRs), all of which are highly conserved in TIR1 proteins of other plant species. Phylogenetic analysis showed that the LeTIR1 protein shared high similarity with other known TIR1 proteins. Both sequence and phylogenetic analysis suggested that LeTIR1 is a TIR1 homologue and encodes an F-box protein in tomato. Semi-quantitative RT-PCR indicated that LeTIR1 was expressed constitutively in all organs tested, with higher expression in stem than root, leaf, flower and fruit. Its expression level was positively correlated with the auxin distribution in stem or axillary shoot, and was induced by spraying exogenous IAA.

The aim of this work was to evaluate the relative gene expression levels of the cytokines IL- 1B, IL-8, IL-12, IFN-γ, IL-4, IL-10 and TGF-β in somatic milk cells of French Alpine breed, anestrous goats that were experimentally infected in the left mammary gland with Staphylococcus chromogenes during the lactation peak. Milk samples were obtained from both glands for 21 consecutive days post infection. Total RNA was extracted, and real-time PCR was conducted using primers specific to each cytokine. The relative RNA expression of the evaluated cytokines was determined by the comparative method 2-ΔΔCT, using milk from the right gland of the goats as a reference (control) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous control. According to the Wilcoxon test results, IL-1B and IL-12 expression levels showed significant differences compared to those in the control group (p<0.05) from 24 hours post infection until the end of lactation; on day three, IL1β, IL8, IL12 and TGF-β had a statistically significant change in expression with respect to those in the control group (p<0.05); closer to the end of the lactation period, there is no overexpression of the anti-inflammatory interleukins (IL-4 and TGF-β) which may reflect the effort of the host immune system to eradicate the microorganism from the mammary gland.

The use of lactoferrin (LF) and/or lactobacillus sp. (LB) to improve animal health and production has increased recently. However, information regarding the immune-modulatory role of LB supplementations either alone or in combination with LF in sheep remains unclear. Therefore, the present study was designed to evaluate the immune modulating properties and the antioxidant activity of supplementing commercially available LF and/or LB in healthy lambs. For this reason, twenty-four apparently healthy Ossimi lambs were used. After three weeks of acclimatization, the lambs were randomly allocated to four equal-sized groups and assigned to receive one of the following supplements: LB at a dose of ~ 1 g active ingredient/head (group 1), LF at a dose rate of 0.5 gm /head (group 2), a combination of both treatments using the same dosing regimens (group 3), and (group 4) received only 10 mL of isotonic saline and was considered as a control group. All supplements were given orally twice daily for 30 consecutive days. Blood samples were collected from each lamb before starting the experiment (T0) and two weeks (T15), and four weeks (T30) after giving supplements for hematological examinations, serum biochemical analyses, and RT-PCR assays. Our findings demonstrated that lambs receiving LB showed statistically significant (P<0.05) higher values of total leucocytes, lymphocytes and lysozyme activity than those receiving LF. In contrast, lambs that received LF had significantly (P< 0.05) higher values of serum catalase, nitric oxide and GSH with a significantly lower MDA level compared with those supplemented with LB. A combination of LF and LB supplementation elicited maximal up-regulation of Tollip, TLR4, IL-5, and IL-6 gene expression compared with other groups. The results suggest that bovine LF and or LB could be used as useful nutritional supplements to support the immune system in healthy lambs.

Onion yellow dwarf virus (OYDV), an aphid-borne potyvirus is one of the major viral pathogens of garlic causing significant yield losses worldwide. It is found almost everywhere in the world where Allium species is grown. The aim of this study was to test the presence of OYDV infection in garlic from Ethiopia. The presence of the virus was tested by Reverse transcription polymerase chain reaction (RT-PCR). The direct sequencing of the PCR product produced a sequence of 296 bp. Sequence analysis showed 89.27% sequence homology with an isolate from Australia (HQ258894) and 89.29% with an isolate from Spain (JX429964). A phylogenetic tree constructed with MEGA 7.0 revealed high levels of homology with various isolates of OYDV from all over the world and thus further confirmed the identity of the virus.

Bovine parvovirus (BPV), bovine coronavirus (BCoV) and bovine parainfluenza virus (BPIV) are common etiologies causing gastrointestinal and respiratory diseases in dairy herds. However, there are few reports on the synchronous detection of BPV, BCoV and BPIV. The present article aimed to develop a quick and accurate RT-PCR assay to synchronously detect BPV, BCoV and BPIV based on their specific probes. One pair universal primers, one pair specific primers and one specific probe was designed and synthesized. After the concentrations of primer and probe and annealing temperature were strictly optimized, the specificity, sensitivity and repeatability of the established triplex probe qRT-PCR were evaluated, respectively. The results showed the recombinant plasmids of pMD18-T-BPV, pMD18-T-BCoV and pMD18-T-BPIV were 554bp, 699bp and 704bp, respectively. The optimal annealing temperature was set at 45.0°C for triplex qRT-PCR. The triplex probe qRT-PCR can only synchronously detect BPV, BCoV and BPIV. Detection sensitivities were 2.0×102, 2.0×102 and 2.0×101 copies/μL for BPV, BCoV and BPIV, being 1000-fold greater than that in the conventional PCR. Detection of clinical samples demon- strated that triplex probe qRT-PCR had a higher sensitivity and specificity. The intra-assay and inter-assay coefficient of variation were lower than 2.0%. Clinical specimens verified that the triplex qRT-PCR had a higher sensitivity and specificity than universal PCR. In conclusion, this triplex probe qRT-PCR could detect only BPV, BCoV and BPIV. Minimum detection limits were 2.0×102 copies/μL for BPV and BCoV, and 2.0×101 copies/μL for BPIV. The sensitivity of this triplex probe qRT-PCR was 1000-fold greater than that in the conventional PCR. The newly qRT-PCR could be used to monitor or differentially diagnose virus infection.

In the spring of 2019, many plants, mainly winter wheat, were observed to have dwarfism and leaf yellowing symptoms. These plants from several regions of Poland were collected and sent to the Plant Disease Clinic of the Institute of Plant Protection – National Research Institute in Poznań to test for the presence of viral diseases. Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) results showed numerous cases of Wheat dwarf virus (WDV) and a few cases of plant infections caused by Barley yellow dwarf viruses (BYDVs). WDV was detected in 163 out of 236 tested winter wheat plants (69.1%), in 10 out of 27 tested winter barley plants (37%) and in 6 out of 7 triticale plants (85.7%) while BYDVs were found, respectively, in 9.7% (23 out of 236) and in 18.5% (5 out of 27) of tested winter forms of wheat and barley plants. Infected plants came mainly from the regions of Lower Silesia and Greater Poland. Furthermore, individual cases of infections were also confirmed in the following districts: Lubusz, Opole, Silesia, Kuyavia-Pomerania and Warmia-Masuria. Results of Duplex-immunocapture-polymerase chain reaction (Duplex-IC-PCR) indicated the dominance of WDV-W form in wheat and WDV-B form in barley plants. Moreover, results of reverse transcription – polymerase chain reaction (RT-PCR) connected with restriction fragment length polymorphism (RFLP) analysis, performed for 17 BYDVs samples, revealed 8 BYDV-PAS, 4 BYDV-MAV and 2 BYDVPAV as well as the presence of two mixed infections of BYDV-MAV/-PAS and one case of BYDV-MAV/-PAV. Next, RT-PCR reactions confirmed single BYDV-GAV infection and the common presence of BYDV-SGV. To the best of our knowledge, in 2020 the viruses were not a big threat to cereal crops in Poland.