Genetic characterization of Indian peste des petits ruminants virus (PPRV) by sequencing and phylogenetic analysis of fusion protein and nucleoprotein gene segments

Peste des petits ruminants (PPR) is an important viral disease of sheep and goats, endemic in India. The study was undertaken to characterize the local PPRV by sequencing fusion (F) protein and nucleoprotein (N) gene segments and phylogenetic analysis, so as to focus on genetic variation in the field viruses. Selected regions of PPRV genome were amplified from clinical samples collected from 32 sheep and goats by RT-PCR and the resulting amplicons were sequenced for phylogenetic analysis. The phylogenetic tree based on the 322bp F gene sequences of PPRV from five different locations clustered them into lineage 4 along with other Asian isolates. While the 425bp N gene sequences revealed a different pattern of branching, yielding three distinct clusters for Nigerian, Turkey and Indian isolates. Thus, classification of PPRV into lineages based on the N gene sequences appeared to yield better picture of molecular epidemiology for PPRV.     

Leukotrienes Affect Secretory Function of Ovarian Cells In Vitro*

The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14–16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC₄, LTB₄, Azelastine (an antagonist of LTC₄) or Dapsone (an antagonist of LTB₄). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5‐lipoxygenase (5‐LO) by real time RT‐PCR, and media were collected for determination of prostaglandin (PG)E₂, F_2α, progesterone (P4; LSC only), endothelin‐1 (ET‐1; LEC only) and 17‐β oestradiol (E2; GC only). The greatest mRNA expression for LTR‐II and 5‐LO were found in LEC, whereas LTR‐I mRNA expression did not differ among cell types. The level of PGE₂ increased after LTs treatment in each type of ovarian cell, excluding LTC₄ treatment in LEC. The secretion of PGF_2α was also increased by LTs, but decreased after LTB₄ treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB₄ stimulated whereas LTC₄ inhibited P4 secretion; in LEC cultures, LTC₄ stimulated but LTB₄ inhibited ET‐1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET‐1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions.     

Litter Size and Vagina–cervix Catheter Penetration Length in Gilts

As in other species, the reproductive tract in pigs increases in size with age and body weight, and the development of the reproductive tract depends on a balance between development of the pituitary–ovarian axis and the influence of metabolic hormones. Two experiments were conducted in prepubertal Duroc gilts, 150–180 days of age, to determine whether litter size is related to vaginal–cervix catheter penetration length during insemination. In experiment 1, oestrus was induced in 452 gilts with a combined dose of 400 IU Pregnant Mare Serum Gonadotrophine (PMSG) + 200 IU human chorionic gonadotropin (hCG). The gilts were classified into three catheter penetration length groups: Ih, ≤ 21 cm; IIh, > 21 and < 28 cm; IIIh, > 28 cm. The litter size was lowest in group Ih (7.35 ± 0.15) compared with groups IIh (7.81 ± 0.12; p < 0.05) and IIIh (10.0 ± 0.36; p < 0.001). In experiment 2, first oestrus was induced in 162 gilts by boar exposure. The gilts were classified into three catheter penetration length groups at insemination during their second oestrus: In, ≤ 24 cm; IIn, > 24 and < 26 cm; IIIn, > 26 cm. As in experiment 1, the litter size was lowest in the group with the shortest catheter penetration length (8.32 ± 0.19). The litter size was not different among gilts of groups IIn and IIIn (8.84 ± 0.35 and 9.56 ± 0.46, respectively), but litter size was lower (p < 0.05) in group In than in group IIn. Based on the combined data from both experiments, the correlation between the catheter penetration length and total number of piglets born was expressed as: y=5.346 ± 0.104x; r=0.361 (p < 0.05). Fertility rate was not different among the groups of gilts induced into oestrus by hormone treatment or inseminated in the second oestrus; however, the total fertility rate of boar‐exposed gilts was higher (p < 0.0001) than PMSG/hCG treated animals. Thus, it is possible to conclude that litter size at first farrowing is associated with vaginal–cervix catheter penetration length during insemination of the gilt.