Molecular identification and genetic characterization of Theileria and Anaplasma infection in sheep and goat of North Gujarat, India.

Hemoprotozoal diseases are significant health concerns in small ruminants. The present study was conducted to identify and characterize the species of Theileria and Anaplasma in sheep and goats located in different districts of North Gujarat, India. A total of 226 (Banaskantha = 175, Patan = 26, and Bhuj = 25) blood samples were collected from sheep (n = 78) and goats (n = 148), and 46 ticks were collected and identified from sheep and goats. PCR assays were carried out using genus and species-specific primers for Theileria targeting 18S rRNA locus and for Anaplasma targeting the msp5 gene. Overall, 37.2% sheep (29/78) and 10.8% of goats (16/148) were positive for Theileria by PCR, whereas 15.4% of sheep (12/78) and 25.7% goats (38/148) were positive for Anaplasma infection. Moreover, mixed infection was found in 4.4% (10/226) of sheep and goats by PCR. Sanger sequencing of Theileria and Anaplasma positives revealed a high similarity to T. ovis and A. ovis using NCBI blast, respectively. Phylogenetic analyses revealed that the Anaplasma spp. DNA sequences belonged to the A. ovis group and closely associated with the A. ovis nucleotide sequence strain Haibei isolated in China from sheep (GQ483471). The phylogenetic analysis based on the SSU rRNA locus revealed that the Theileria ovis DNA sequences belonged to the T. ovis group and closely related to MW440586 isolated in Kerala, India, from a goat. The majority of ticks (91.3%) were identified as Hyalomma. In conclusion, Theileria ovis and Anaplasma ovis were commonly identified species in sheep and goats and transmitted mainly by Hyalomma ticks in North Gujarat, India, which is important baseline data for future research and control strategies. This is the first report on Theileria and Anaplasma co-infections in sheep and goats from North Gujarat, India.

Doppler indices of uterine arteries and mRNA transcripts of hormone receptors in cervix vis-à-vis dilatation therapy in buffaloes affected with incomplete cervical dilatation associated with uterine torsion.

The present study aimed to investigate the Doppler indices and mRNA transcripts of hormone receptors in relation to the response of dilatation therapy in incomplete cervical dilatation (ICD) associated with uterine torsion in buffaloes. Out of 36 successfully detorted uterine torsion cases, eight buffaloes revealed a fully dilated cervix, while the remaining 28 had ICD, and subjected to dilatation therapy (500 µg cloprostenol + 2 mg estradiol benzoate + 80 mg valethamate bromide + 50 IU oxytocin + 250 mL calcium borogluconate). The responses of dilatation therapy were assessed in 26 buffaloes as one died, and one could not follow up. Doppler indices of middle uterine arteries on trans-rectal ultrasound were evaluated pre- and 30-60 min post-detorsion. Cervical tissue biopsies were collected from 16 buffaloes to study mRNA transcripts of hormone receptors. The duration, degree, location of uterine torsion, fetal viability, consistency of the cervix, relaxation of pelvic ligaments, udder engorgement, and gestation length were also recorded to evaluate the response of dilatation therapy. The 73.08% (19/26) buffaloes responded to the therapy with a duration ranging from 2 to 56 hrs (18.41 ± 4.11). The significantly increased blood flow volume (BFV) and time-average peak velocity (TAP) while the significantly reduced resistive index (RI) and pulsatility index (PI) in an ipsilateral middle uterine artery (MUA) at post-detorsion were observed in dilation therapy responded than the not-responded group. The mRNA transcripts of estradiol receptors-α (ESR1), prostaglandin receptors (PTGFR), and oxytocin receptors (OXTR) were upregulated by 7.47, 6.63, and 8.72-fold in the ICD group, respectively. The Doppler indices along with duration of illness, location of uterine torsion, consistency of the cervix, and udder engorgement can be used to predict the response of dilatation therapy in ICD associated with uterine torsion. The upregulated mRNA expression of ESR1, PTGFR and OXTR is mandatory for success of dilatation therapy.

Effect of dietary n-3 polyunsaturated fatty acid rich fish oil on the endometrial prostaglandin production in the doe (Capra hircus).

Recently, we showed that dietary supplementation of n-3 PUFA rich fish oil (FO) decreased the metabolites of serum prostaglandin (PG) F2α and E2 during the window of pregnancy recognition in the doe. In this study, we investigated its effect on the changes on endometrial PG production in vitro. Cycling does (n = 12) of Rohilkhand region were divided into two equal groups and fed a concentrate diet supplemented with either FO containing 26% n-3 PUFA (TRT; n = 6) or palm oil (CON; n = 6) @ 0.6 mL/kg body weight for 57 days. Estrus was synchronized by two injections of PGF2α analogue viz, on day 25 and 36 of supplementation and laparo-hysterotomy was performed to obtain endometrial tissue on day 16 of the synchronized estrus. Endometrial explant culture was done using a defined medium.The basal PG production was assayed at 6 and 12 h. Endometrial explant was stimulated with oxytocin (OXT) and/or recombinant ovine interferon tau (roIFN-τ) and PGs were assayed at 3 and 12 h post-treatment. The relative expression of genes related to PG metabolism in the endometrium was done by Quantitative Real Time PCR technique (qRT-PCR). There was a significant (P < 0.05) decline in the basal production of PGF2α and PGE2 in the TRT as compared to the CON group. The cultured endometrial tissue produced PGF2α in a time- dependent fashion in both the groups (P < 0.05). Neither OXT nor roIFN-τ had a significant (P > 0.05) effect on the PGF2α and PGE2 production in the TRT group. Similarly, the PG production in the OXT and roIFN-τ was comparable with the control in TRT. Expression of mRNA for cyclooxygenase-2 (COX-2), cytosolic phospholipase A2 (cPLA2) and PGF synthase (PGFS) was lower (P < 0.05) whereas, PGE synthase and peroxisome proliferator-activated receptors such as PPAR-γ and δ was increased (P < 0.05) in n-3 PUFA fed doe. In conclusion, dietary supplementation of FO decreased the endometrial production of PGF2α and PGE2 by downregulating the COX-2, cPLA2 and PGFS transcripts in the doe. The findings suggest that n-3 PUFA influence embryo survival by modulating the endometrial PG.

Effect of dietary supplementation of omega-3 polyunsaturated fatty acid (PUFA) rich fish oil on reproductive performance of the goat (Capra hircus).

Dietary supplementation of n-3 PUFA decreases the luteolytic PGF2α and improves the pregnancy rate in the dairy cow. However, its effect in the goat is not known. Accordingly, we studied the effect of supplementation of n-3 PUFA rich Fish oil (FO) on different reproductive events in the goat. Cycling goats (n = 30) were divided into two equal groups and fed an isocaloric and isonitrogenous diet supplemented with either FO (TRT; n = 15) or palm oil (PO) (CON; n = 15) @ 0.6 mL/kg body weight for 72 days during the breeding season. Estrus synchronization was done on day 25 and 36 of supplementation using two PG regimen and the goats in estrus were bred. Mean interval from PGF2α administration to the onset of estrus was 12 h longer (P < 0.05) in the TRT group than that of CON. The number of preovulatory follicles (POF) and ovulation rate was significantly higher in FO supplemented goats (P < 0.05) by 39.64 and 41.35%, respectively. Though the corpus luteum diameter was significantly higher (P < 0.05) on day 5, 8 and 11 post-breeding in the TRT group, mean serum progesterone (P4) did not differ significantly (P > 0.05). Mean concentration of serum estradiol (E2) was significantly (P < 0.01) lower in the FO supplemented group during day 0-60 post-breeding which could be due to significantly low serum cholesterol (P < 0.01). Though the serum concentration of PGF2α metabolite (PGFM) and PGE2 metabolite (PGEM) in the pregnant goats was significantly (P < 0.05) lower in the TRT group on day 16 and 17 post-breeding, the ratio of PGEM to PGFM remained unaffected suggesting a favourable effect of FO supplementation on the early pregnancy. The number of embryos, twinning rate and kidding rate were high in FO supplemented group though it was non-significant. However, gestation length, birth weight of kids and neonatal behaviour were comparable between the groups (P > 0.05). In conclusion, supplementation of n-3 PUFA rich FO significantly increased the number of POF and ovulation rate with numerical increase in the kidding rate. Further, it decreased the serum E2 and PGFM during the critical window of pregnancy recognition in the doe.