Oleuropein activates autophagy to circumvent anti-plasmodial defense.

Antimalarial drug resistance and unavailability of effective vaccine warrant for newer drugs and drug targets. Hence, anti-inflammatory activity of phyto-compound (oleuropein; OLP) was determined in antigen (LPS)-stimulated human THP-1 macrophages (macrophage model of inflammation; MMI). Reduction in the inflammation was controlled by the PI3K-Akt1 signaling to establish the "immune-homeostasis." Also, OLP treatment influenced the cell death/autophagy axis leading to the modulated inflammation for extended cell survival. The findings with MII prompted us to detect the antimalarial activity of OLP in the wild type (3D7), D10-expressing GFP-Atg18 parasite, and chloroquine-resistant (Dd2) parasite. OLP did not show the parasite inhibition in the routine in vitro culture of P. falciparum whereas OLP increased the antimalarial activity of artesunate. The molecular docking of autophagy-related proteins, investigations with MMI, and parasite inhibition assays indicated that the host activated the autophagy to survive OLP pressure. The challenge model of P. berghei infection showed to induce autophagy for circumventing anti-plasmodial defenses.

Seroprevalence of Bovine ephemeral fever virus in Gujarat State of India.

Bovine ephemeral fever (BEF) virus (BEFV) is an arthropod borne virus that causes bovine ephemeral fever or three­day sickness in cattle and buffaloes. This is the first report on seroprevalence of BEF in cattle and buffaloes in Gujarat, India. Total of 92 animals, 78 cattle and 14 buffaloes from three regions (districts) of Gujarat state of India, were screened for the presence of anti­BEF antibodies. A total of 27 out of 92 animals were found positive and overall seroprevalence detected was 29.34% (95% CI 20.0­38.6%). A total of 19 out of 78 cattle and 8 out of 14 buffalo's samples were found positive BEFV antibodies. Species­wise seroprevalence in cattle and buffaloes was 24.35% (95% CI 14.8­33.8%) and 57.1% (95% CI 31.2­83.0%), respectively. There was a statistically significant (p < 0.05) species effect based on the seroprevalence. In cattle, location­wise seroprevalence was observed to be 26.82% (95% CI 13.2­40.3%) and 21.62% (95% CI 8.3­34.8%) in Navsari and Banaskantha districts, respectively. The effect of location is not statistically significant (p < 0.05). Cytopathic effect of Vero cells was characterized by rounding, granulation of the cytoplasm within 48­72 hrs of post infection. This was the first report demonstrating the presence of BEFV in Gujarat state.

VP2 gene sequencing based Geno-grouping of infectious bursal disease viruses isolated from Gujarat and Maharashtra state (India).

Infectious bursal disease (IBD), caused by infectious bursal disease virus (IBDV), has recently been reported in chickens vaccinated with classical or intermediate types of vaccines from various regions of India due to the emergence of novel very virulent strains of infectious bursal disease virus (vvIBDV). In the present study, suspected samples of IBD were collected from poultry flocks of districts of Gujarat and Nagpur (Maharashtra), identified using PCR and grouped as per traditional and new genogrouping pattern. Out of 54 bursa samples, 21 (38.89%) yielded the expected amplicon of 743 bp (701-1444 bp), and were found positive for IBDV. Among these 21 positive flocks, 11 (52.38%) were already vaccinated. Upon nucleotide sequencing of amplicon and its deduction into amino acids, it was found that all the sequences of present study were related to vvIBDV according to old classification pattern. Considering the new genogrouping pattern, nine and four sequences of this study fell within G3a and G3b lineage, respectively. These sequences revealed important differences at key amino acid positions with respect to classical (G1 genogroup), variant (G2 genogroup) type of IBDV and classical vaccines. Further divergence from prototypic vvIBDV strains was revealed as, D-N at 212 position (N = 9) and 279 position (N = 1). In sequences from Maharashtra (group 2 of G3a lineage), occurrence of V instead of P/T/A at 222 position was recorded as a novel and conspicuous substitution in the immunodominant peak A of VP2 hypervariable region. Additional changes at 270 (3 sequences) and 272 positions (4 sequences) could be attributed to reverse mutation or recombination with vaccine strains. In conclusion, both point mutation and genetic reassortment with intermediate type of vaccines were found to be responsible for generation of novel vvIBDV strains in this area which belonged to G3a and G3b genogroups.

Genotyping of Indian antigenic, vaccine, and field Brucella spp. using multilocus sequence typing.

INTRODUCTION: Brucellosis is one of the most important zoonotic diseases that affects multiple livestock species and causes great economic losses. The highly conserved genomes of Brucella, with > 90% homology among species, makes it important to study the genetic diversity circulating in the country. METHODOLOGY: A total of 26 Brucella spp. (4 reference strains and 22 field isolates) and 1 B. melitensis draft genome sequence from India (B. melitensis Bm IND1) were included for sequence typing. The field isolates were identified by biochemical tests and confirmed by both conventional and quantitative polymerase chain reaction (qPCR) targeting bcsp 31Brucella genus-specific marker. Brucella speciation and biotyping was done by Bruce ladder, probe qPCR, and AMOS PCRs, respectively, and genotyping was done by multilocus sequence typing (MLST). RESULTS: The MLST typing of 27 Brucella spp. revealed five distinct sequence types (STs); the B. abortus S99 reference strain and 21 B. abortus field isolates belonged to ST1. On the other hand, the vaccine strain B. abortus S19 was genotyped as ST5. Similarly, B. melitensis 16M reference strain and one B. melitensis field isolate were grouped into ST7. Another B. melitensis field isolate belonged to ST8 (draft genome sequence from India), and only B. suis 1330 reference strain was found to be ST14. CONCLUSION: The sequences revealed genetic similarity of the Indian strains to the global reference and field strains. The study highlights the usefulness of MLST for typing of field isolates and validation of reference strains used for diagnosis and vaccination against brucellosis.