Comparative study of reproductive and estrus characteristics of sexed and conventional semen in crossbred cows under field conditions.

Background and Aim: Sexed semen (SS), a reproductive biotechnology tool, can alter the sex ratio of offspring in bovines. This study elucidates a comparative analysis of estrus-related parameters influencing conception rate and pregnancy losses under field conditions between conventional and SS. Materials and Methods: In the present study, artificial insemination with (SS; n = 143) and conventional semen (CS; n = 143) was performed at spontaneous estrus, i.e., 16-18 h after the onset of estrus signs, to analyze their comparative evaluation in terms of conception rates in crossbred cows under field conditions. Different parameters such as age, parity, body condition score (BCS), estrus duration, inter-estrus interval (IEI), diameter of pre-ovulatory follicle (DPOF) at estrus, and cervical mucus properties (pH and spinnbarkeit [SBK]) were recorded for each cow. Results: The first insemination conception rates for sexed and conventional semen were 55.24% and 63.63% whereas the overall conception rates were 49.14% and 57.37% on days 35 and 75 post-insemination, respectively, with no significant difference (p > 0.05). Conception rates between sexed and CS inseminations were statistically significant (p < 0.01), whereas factors such as age, parity, BCS, DPOF, IEI), and SBK value exhibited no substantial variance (p > 0.05) for both types of semen straw. Conclusion: SS straws yielded a conception rate equivalent to CS straws, with estrus duration being the key factor affecting conception under field conditions.

Comprehensive exploration unveiling the sonography and histopathology of uterine leiomyoma in a cow.

Genital tumours are rare among cattle, largely due to their relatively short lifespans. Leio-myoma, a smooth muscle tumour being more prevalent in dogs, appears only at a rate of 1.00 - 2.00% in cattle, affecting reproductive efficiency in cases of complete uterine obstruction. This case report involves an 8-year-old cow with repeated insemination attempts unveiled 5.00 cm intra-luminal uterine mass, obstructing the right uterine horn. Transrectal sonography (TRUS) revealed a highly vascularized mass with normal ovarian function. Confirmation of clinical condition, i.e., uterine leiomyoma, via uterine biopsy concluded the presence of neoplastic smooth muscle cells arranged in interlacing bundles showing mild pleomorphism, and special staining using Masson's trichrome revealed an unappreciable amount of connective tissue; subsequently right flank celiotomy was performed to remove the benign tumour. Forty-five days after celiotomy, TRUS examination confirmed an unobstructed uterine horn, and bilateral oviduct patency was adjudged with 2.50% methylene blue. Following treatment for chronic endometritis, artificial insemination led to conception nearly 90 days post-procedure. The TRUS aids preliminary diagnosis, while definitive identification demands necropsy and surgical methods. This case underscores the diagnostic significance of TRUS, histopathology and celiotomy for identifying and managing uterine leiomyoma in cattle.

Temporal alteration in ovarian follicle fate with hormonal and hemodynamic perspectives during receptive phase of estrous cycle in endometritic cattle.

The present research was carried out to assess the serum progesterone (P4) concentration and uterine hemodynamics at estrus till ovulation in cyclic cows (N = 130) with healthy or diseased uterus. At estrus, 85 cows were diagnosed with clinical endometritis (CE; n = 44) and sub-clinical endometritis (SCE; n = 41); whereas, 45 cows being served as control namely no endometritis (NE; n = 45) were included in the study. Serum progesterone estimation at 12 - 14 and 40 hr after the onset of estrus and Doppler sonography of both middle uterine arteries were done to envisage the uterine hemodynamics and ovulation. The serum progesterone concentration was significantly higher at 12 - 14 hr after onset of estrus in CE and SCE cows. At 12 - 14 hr after onset of estrus, a cut-off value of ≥ 0.48 ng mL-1 P4 was obtained, above which 22.72% CE, 26.82% SCE and only 8.88% NE cows failed to ovulate within 36 - 40 hr of estrus onset. Among the Doppler indices, pulsatility and resistance indices were significantly higher; whereas, volume and velocity indices were significantly lower in NE cows. In cows diagnosed with CE and SCE, a higher supra-basal P4 concentration, and velocity and volume of blood flow to uterus at estrus negatively affected the duration to ovulation.

Temporal disposition of pelvic girdle, vagina and vulva in relation to urovagina in cycling and pregnant-postpartum cows.

Urovagina (UV) is less studied in cows. The vaginal contents, constrictor vestibule muscle activity, circulating progesterone, pelvic girdle, vagina and vulva angles were compared on Days 0 and 14 within and between UV (UV group; n = 36) and normal (N group; n = 36) cows. The oestrous duration was compared among the groups. Parameters for various UV severities were also compared. Another set of pregnant-postpartum cows (P-PP group; n = 31) underwent monthly evaluations for UV and the angles from the third month of pregnancy until the second month postpartum. The effect of age, parity and body condition score on UV severities on Day 0, and parity on angles in the P-PP group was evaluated. Different variables were correlated in different groups. The UV group was repeat breeder, exhibited prolonged oestrus and reduced progesterone on Day 14. The latter increased with UV severity on Day 0. On Day 14, severe form of UV was more prevalent. The UV severity increased with parity. In the UV group, a cranioventral pelvic girdle on Day 0 became more cranioventral on Day 14 and was correlated with the vagina moving from a caudodorsal position on Day 0 to a cranioventral position on Day 14. In the P-PP group, the pelvic girdle and vagina remained caudodorsal and UV was undetectable throughout the study. Vagina and vulva displayed a parity-by-month interaction in the P-PP group. To summarize, the UV characteristics were influenced by the oestrous cycle stage, pelvic girdle angles, and parity.

Additional intracervical prostaglandin reduces induction to calving time in buffaloes affected with incomplete cervical dilatation post-uterine detorsion.

Conventional induction protocol (CIP) of calving in buffaloes employs the intramuscular (IM) administration of dexamethasone (40 mg) and cloprostenol sodium (500 µg). If there is no progression in terms of cervical dilatation, then a second dose of cloprostenol sodium (500 µg) is administered intramuscularly. This protocol possesses certain demerits: (1) a wide range of response time intervals, and (2) increased risk of fetal membrane retention. Considering the cervix as a caudal continuation of the myometrium with its own contractile potential, and the limitations of CIP, we developed intracervical (IC) drug administration route in buffaloes. The proposed technique was evaluated for its use in a total of 22 cases of incomplete cervical dilatation in uterine torsion-affected buffaloes (IC-14 and IM-8). In addition to CIP, the IC group received an intracervical injection of cloprostenol sodium (500 µg) at the start of the experiment whereas the IM group received an extra intramuscular dose of cloprostenol sodium (500 µg) either after 24 h or when no progression in cervical dilatation is noticed. Surprisingly, the average response time during the experiment in the IC group was 5.8 h shorter (p < 0.000) than in the IM group (IC-5.7 ± 0.17 h vs. IM-11.9 ± 0.74 h). The duration from calving to fetal membrane expulsion (IC-12.8 ± 0.60 h vs. IM-17.5 ± 1.40 h; p < 0.002) and incidence of retention of fetal membrane were also less in the IC group (57.1% vs. 87.5%). The proposed intracervical drug administration potentiates cervical dilatation and can be regarded as a safe, effective, and feasible technique for attaining reliable results.

Effect of ovarian follicular wave pattern and endocrine characteristics on pregnancy outcome in cows.

The effect of two (2W) versus three (3W) wave patterns of follicular dynamics and concurrent endocrine milieu of follicle-stimulating hormone (FSH), luteinizing hormone (LH), oestradiol 17-ß (E2) and progesterone (P4) were investigated during one interoestrous interval (IEI) before insemination, on ensuing pregnancy, in 70 lactating Jersey crossbred cows. The findings were evaluated for between [included all (overall) 2W-O and 3W-O cows] and within [after separating pregnant (P) and non-pregnant (NP) cows in 2W and 3W] wave patterns. The propensity of two (58.6%, n = 41) and three (41.4%, n = 29) wave patterns was similar (p = .15). The IEI, shorter by 2.6 days for 2W-O versus 3W-O (p < .0009), predicted wave pattern as 100% 2W-O cows had IEI of ≤21 days, present only in 27.6% 3W-O cows (p < .0001). The ovulatory follicle persisted for a significantly shorter duration for 3W-O versus 2W-O cows. The average FSH, LH, E2 and P4 concentrations during the IEI did not differ for between and within the wave patterns. Pregnancy rate (%) of 58.6 versus 41.4 (p = .15) for 2W-O versus 3W-O and 56.1-P versus 43.9-NP (p = .44) for within 2W was similar, but tended to differ for within the 3W pattern (69.0-P versus 31.0-NP, p = .06). The pregnancy outcome was influenced by the age of ovulatory follicle for between the wave patterns and by follicular count as well as FSH surge concentration for within the wave patterns. A shorter luteal phase reduced the pregnancy outcome, a novel finding of the present study.

Proteome dataset of pre-ovulatory follicular fluids from less fertile dairy cows.

This article contains raw and processed data related to research published in Zachut et al. (2016) [1]. Proteomics data from preovulatory follicles in cows was obtained by liquid chromatography-mass spectrometry following protein extraction. Differential expression between controls and less fertile cows (LFC) was quantified using MS1 intensity based label-free. The only previous proteomic analysis of bovine FF detected merely 40 proteins in follicular cysts obtained from the slaughterhouse (Maniwa et al., 2005) [2], and the abundance of proteins in the bovine preovulatory FF remains unknown. Therefore, the objectives were to establish the first dataset of FF proteome in preovulatory follicles of cows, and to examine differentially expressed proteins in FF obtained in-vivo from preovulatory follicles of less fertile cows (also termed "repeat breeder") and control (CTL) cows. The proteome of FF from 10 preovulatory follicles that were aspirated in vivo (estradiol/progesterone>1) was analyzed. This novel dataset contains 219 identified and quantified proteins in FF, consisting mainly of binding proteins, proteases, receptor ligands, enzymes and transporters. In addition, differential abundance of 8 proteins relevant to follicular function was found in LFC compared to CTL; these findings are discussed in our recent research article Zachut et al. (2016) [1]. The present dataset of bovine FF proteome can be used as a reference for any study involving disorders of follicular development in dairy cows or in comparative studies between species.

Proteomic analysis of preovulatory follicular fluid reveals differentially abundant proteins in less fertile dairy cows.

UNLABELLED: The follicular fluid (FF) proteome can provide an indication of follicular quality. High-yielding dairy cows suffer from low fertility, which could be related to follicular function. However, the proteome of preovulatory follicles has never been described in cows. Our objectives were to: 1) define the bovine preovulatory FF proteome, and 2) examine differentially abundant proteins in FF of controls (CTL, n=10) and less fertile cows (LFC; failed to conceive following ≥6 inseminations, n=8). Follicles ≥7mm in diameter were aspirated in vivo, and estradiol (E2) and progesterone (P4) were examined. The FF from 10 preovulatory follicles (E2/P4>1) was analyzed; E2 was higher and follicle diameter tended to be larger in LFC. As aspirations were conducted at a fixed time, this suggests accelerated follicular growth in LFC. The 219 identified and quantified proteins consisted mainly of binding proteins, proteases, receptor ligands, enzymes and transporters. Differential abundance of 8 relevant proteins was found in LFC compared to CTL: SERPINA1, TIMP2, ITIH1, HSPG2, C8A, COL1A2, F2, and IL1RAP. These proteins could influence follicular function-e.g., decreased SERPINA1 may be related to accelerated follicular growth-and therefore, further examination of their roles in the etiology of LFC is warranted. SIGNIFICANCE: High yielding dairy cows suffer from infertility that leads to major economic losses worldwide. In Israel, about 30% of dairy cows fail to conceive following ≥4 inseminations. The etiology of this low fertility is multifactorial and remains a serious challenge. Follicular fluid proteome can provide indication to follicular quality, yet the proteome of pre-ovulatory follicles has not been described in cows. This work examined the differential abundance of proteins in less fertile dairy cows compared to controls, and found 8 relevant novel proteins that could influence follicular function. The role of these proteins in the etiology of less fertile cows should be further examined.

Polymorphonuclear neutrophil profile in genital discharge vis-à-vis pregnancy outcome in crossbred cows: effect of varying cow fertility and insemination from different bulls.

The study investigated the relationship between count and/or viability percentage of polymorphonuclear neutrophil(s) (PMN) in genital discharge and pregnancy outcome during 180 estrous periods in 163 normal and repeat breeder. The relationship was also evaluated as a response to AI by three of the most frequently used bulls. The estrous periods leading to pregnancy in the normal cows were considered fertile (FE; n=66), whereas others with pregnancy failure were considered non-fertile in the remaining normal (NFE; n=86) and the entire group of repeat breeder cows (RB; n=28) cows. Another set of reproductively healthy cows, not inseminated, were also studied (C; n=10). The PMN characteristics were investigated in 3µl fraction of cervical discharge, aspirated initially at mid-estrus in C/pre-AI in the inseminated cows and subsequently 18 to 24h later. The first sample did not reveal PMN in all the cows. However, there was a variable PMN response in the second sample. The PMN count of 9.3±1.9 in the C group was least and significantly less than the FE and NFE groups. A count of 90.9±8.4 in the FE group was greater (P<0.01) than 60.9±5.1 in the NFE and 36.7±5.2 in the RB group. The PMN viability percentage among the four groups was similar (range: 61.3±6.5% to 70.1±6.4%). The average PMN count in response to AI with all the three bulls, JY-426 (47 inseminations), JY-450 (23 inseminations) and HF-5 (33 inseminations), was less in the NFE than the FE group, the differences being marked in NFE compared with FE for JY-450 (51.8±12.4 compared with 88.0±17.4; P=0.10) and HF-5 (38.5±10.1 compared with 92.9±26.5; P<0.05). It is concluded that a greater PMN count post-AI could favor pregnancy outcome. Further, PMN migration is likely to be controlled by maternal factors rather than semen characteristics.

Population genetic structure of the melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) based on mitochondrial cytochrome oxidase (COI) gene sequences.

Population genetic structure of melon fly analysed with mitochondrial cytochrome oxidase I gene suggested that melon fly populations across the globe is homogeneous with non-significant variation of 0.000-0.003 base substitutions per site. Test isolates representing various geographic situations across the world were placed in 26 mitochondrial haplotypes based on variations associated with a maximum of three mutational steps and the predominant haplotype i.e. H1 was present in all melon fly populations except Hawaiian population. Evolution of mtCOI gene suggested that the fly could have originated some 0.4 million years ago. The present study also indicated that the B. cucurbitae population expansion is an event of post Pleistocene warm climatic conditions with small number of founder population. The invasion of B. cucurbitae in Hawaii was associated with the large population size and the global presence of the fly is associated with human mediated dispersal. The very low genetic variation suggested that the fly management might be possible by large scale sterile insect techniques programme.

Molecular and biochemical characterization of three bacterial symbionts of fruit fly, Bactrocera tau (Tephritidae: Diptera).

Characterization of three bacterial symbionts (BC1, BC2 and BC3) of fruit fly Bactrocera tau including morphological, biochemical and 16S rDNA (rrs gene) analysis was done to determine their taxonomic position. Morphological and biochemical characterization placed two bacteria (BC1, Klebsiella oxytoca and BC2, Pantoea agglomerans) into family Enterobacteriaceae and the third one (BC3, Staphylococcus sp.) into family Staphylococcaceae. 16S rDNA gene sequence comparison with the available NCBI database sequences further confirmed the characterizations of bacterial symbionts. Molecular phylogeny of Klebsiella oxytoca and Pantoea agglomerans closely related to the other free living enterobacterial members with 74 to 93% sequence homology (genetic distance 0.000 to 0.085); however, they showed only 74 to 87% similarity with other insect symbionts (genetic distance 0.090 to 0.121). Staphylococcus sp. showed 94% sequence homology with other members of family Staphylococcaceae with the genetic distance of 0.013. Population of these symbionts in adult fruit flies increased exponentially up to the 10th day of adult emergence and thereafter it became almost constant.

Effect of lameness on estrous behavior in crossbred cows.

Lameness, a common disease in crossbred dairy cows, badly affects their fertility. To elucidate, if estrous behavior, comprising of duration and expression of certain estrous behavioral sign(s) (EBS), is involved, an estrous expression score system was applied to objectively compare estrous behavior during 27 and 58 estrous periods, respectively, in 17 normal and 37 lame cows kept in a group of 200 cows under loose housing system in tropical India. The cows were detected in estrus and were closely watched daily for 30 min at 0630, 1030, 1430 and 1830 h from the beginning to the end of estrus. The duration of estrus was similar in the normal and the lame cows (18.7+/-1.2 h versus 17.2+/-0.9 h) and almost similar EBS were shown by two types of cows, albeit with varying frequency. Standing to be mounted, the highest ranking EBS was shown by equal proportions of the cows in both the groups (76.4% versus 73.0%). A lower proportion of lame cows showed head side mounting (35% versus 53%; P<0.05). The frequency of standing to be mounted events was lower in the lame than in the normal cows (2.4+/-0.4 versus 8.0+/-2.5; P<0.05). The lame cows earned non-significantly lower than normal estrous behavior score (1100+/-179 versus 1610+/-357). Further, the presence of more than one cow simultaneously in estrus enhanced the estrous behavior score in the normal (406.4+/-63.0 for one cow in estrus versus 793.0+/-146.0 for three cows simultaneously in estrus; P<0.05), but not in the lame cows. Lameness appeared to have suppressed the playful behavior during estrus probably owing to pain related stress. It is concluded that mildly lame cows appear to have normal physiological estrus. However, some of the EBS remain subdued which prone them to remain undetected for timely breeding.