Introduction

Inactive ovaries, a reproductive problem in which the ovaries are small, smooth, and related with losses structures in ovary ( 1 ). So, the cow lacks signs of estrous and when examined by rectal palpation for diagnosis found small size ovaries with smooth surface and sometime found small follicle present ( 2 ). For this reason, the activity of ovary insufficient to produce gonadotropin hormone, this hormone necessary to induced follicle development, also sometime failure of ovary to respond to gonadotropin, and this condition association dairy cow with highly yield occasionally in heifer in first calving ( 3 ). In this field multiple hormones used to estrous induction when the cow diagnosis anestrous, the commune hormones used include (GnRH) gonadotropin releasing hormone, (FSH) follicle stimulating hormone, (LH) luteinizing hormone and (E2) estradiol hormone ( 4 and 5 ). Some recent studies also encourage the use of progesterone to treatment the inactive ovaries in cow ( 6; 3 ). On the other hand, the use of medicinal plants has contributed to the treatment of many conditions related to public health and reproduction in worldwide ( 7and78 ). The Lepidium sativum (LS), usually identified as cress garden, is a yearly concrescence, comestible, the herb fast growing which belongs to Brassicaceae family ( 9 ). The LS seeds contained multiple of nutrient component protein, carbohydrate, fat, phosphorous, potassium, magnesium, sulfur, calcium sodium, copper, iron, zinc and manganese, other contained the non-essential amino acid (glutamic acid, aspartic acid, serine, glycine, tyrosine, alanine and proline also contains essential amino acid (arginine, histidine, threonine, phenyl alanine, methionine, leucine, isoleucine, and lysine) ( 10 ). The LS herb extract used to treatment of different human aliment such as man seminal weakness ( 11 ), fracture healing ( 12 ), liver disorders ( 13 ). The herb extract has high inhibitory effect agent different pathogen organisms such as Escherichia coli, Staphylococcus aureus, Proteus vulgaris and Klebsiella pneumonae ( 14 ). In female LS seed extract used in different breeding aliment such as inducing abortion ( 15 ) , used contraceptive( 16 ) and increasing milk production ( 17 ),. But to date no study investigates the effects of seed extract to LS herb on ovarian activity. So, this study aimed to compare between effect of seed extract to LS herb and hormonal treatments to inactive ovaries in cows

Materials and methods

Ethical approval: The study has been accepted by the Depart. of Surgery and Therio., College of Vet. Med., Uni, of Baghdad Iraq.

Study period and location: This study was conducted on 27 lactating anestrus crossbreed cows, their age between 3-5 years old in Al- Thahab Al-Abiad village / Abu-Ghriab/ Baghdad province during June 2020-July 2021.

Animals: collected Blood was purified using chloroform phenol ( 18 ). Using Roche's 1st Strand cDNA Synthesis Kit for RT-PCR (AMV), 0.5-1.5 µg of total RNA was used to synthesize single-strand cDNA and a random primer. ( 19 )

PCR gene expression: The cycle RT consists of ten minutes of annealing at 25 degrees Celsius, sixty minutes of cDNA synthesis at 42 degrees Celsius, and five minutes of inactivation at 99 degrees Celsius. I had Electrophoresis done with DNase with a commercial kit (Promega, USA). These animals were suffered from ovarian inactivity after 60-90 days postpartum and diagnosed by rectal palpation and ultrasonography. These cows divided randomly into three equal groups (9 cows on each). The 1^st group treated by GnRH (Receptal®- Ceva) injected IM with 0.021 mg/animal (5ml), the 2^nd group treated with Lepidium sativum (The seeds were ground by an electric mixer into a fine powder, and then the powder was kept in a clean container in the refrigerator at a temperature of 4°C until used in the experiment , then the seeds powder were mixed with distal water at the time of treatment (suspension) in dose 25 mg/orally/ for 10days (250mg/animal), while the 3^rd group considered as control group (without treatment).The reproductive parameters include animal response, the time it takes of response, services number per conception, rate of pregnancy and days open were recorded ( 4 ).

Statistical analysis: The statistical analysis includes the mean, the standard error, the Chi- Square, the T test and variance analysis were used according to ( 18 ).

Results and discussion

The results are shown in table -1 explain the number of animal response were 7 (77.7%), 6 (66.6%) and 3 (33.3%) in the 1^st, 2^nd and 3^rd group respectively with a highly significant different in (P<0.01) for the 1^st (GnRH treated

group) and 2^nd (LS treated group) in compare with the 3^rd group (control), The duration of response recorded 6.36±1.24, 13.72±2.45 and 87.2±0.1in the 1^st, 2^nd and 3^rd group respectively, recording a highly significant differences (P<0.01) for the treatment groups (G1, G2) when compare with control group (G3) and for G1 compared with G2, the No. of services/conception showed no significant differences (P<0.01) in the three groups respectively 1.32±0.7 , 1.25±0.3 and 1.5±0.0.

Pregnancy rate (table 2) was 85.7%, 83.3% and 66.6% in the three groups respectively with a highly significant difference in (P<0.01) for the 1^st and 2^nd group when compare with the 3^rd group, and days open recorded 106.35±5.47, 114.26±6.52 and 187.40±0.0 in the 1^st, 2^nd and 3^rd group respectively which was high significantly in (P<0.01) in treatment groups than the control group. In conclusion: the use of LS seed extract was effective in treated ovarian inactivity in lactating anestrus crossbreed cows close to GnRH. The clinical roles of UHPLC-QTOF-MS were further investigated. TCGA data showed that expression of UHPLC-QTOF-MS in treatment by Lepidium sativum was significantly higher than control (fig. 1)

Figure 1.FAM channel fluorescence is determined by the cycle number.

Theresults of table 1 recorded a highly significant different in (P<0.01) in animal response for the 1^st (GnRH treated group) and 2^nd (LS treated group) in compare with the 3^rd group (control) and this agreed with ( 20 ), ( 7 ) who used a different herbal extract for treatment of inactive ovaries in cows achieving 80% response rate and it agreed with ( 21 ) when used LS seed supplement with intake food , observed the herb lead to increase food intake and increase secretion of gonadotropins hormone in rabbit When the gonadotropin mediated through LS seeds activated the estrogento resulting increase of FSH and LH hormone secretion. Also, it agreed with ( 22 , 23 and 24 )who recorded 50%; 66.6% and 50% respectively to response to GnRH treatment of ovarian inactivity in cows and buffaloes, the result studies explain the systemic administration of GnRH hormone causes to increase FSH and LH levels in plasma in cows ( 2 , 4 ), then these hormones stimulated development of follicles in the ovary and induction of estrous signs in female ( 23 , 24 and 5 , 6 ). The duration of response recording a highly significant differences (P<0.01) for the treatment groups (G1, G2) when compare with control group (G3) and for G1 compared with G2 and these results agreed with ( 22 ) , ( 25 ) and with ( 26 ), ( 21 ), the number of services/conception showed no significant differences in the three groups respectively and this agreed with ( 22 , 26 ). Pregnancy rate (table 2 ) recording a highly significant difference in (P<0.01) for the 1^st and 2^nd group when compare with the 3^rd group, and days open recorded high significantly (P<0.01) in treatment groups than the control group, Increased expression of UHPLC-QTOF-MS Metabolite (fig. 1, 2) these results agree with ( 27; 28)

Figure 2.The gene expression of UHPLC-QTOF-MS Metabolite Extraction in cows

Conclusion

The use of LS seed extract was effective in treated ovarian inactivity in lactating anestrus crossbreed cows and it is similar to the effect of hormonal therapy with fewer negative aspects that may occur during hormone treatment.

Acknowledgments

This study supported by College of Veterinary Medicine, University of Baghdad.

Novelty Statement

This study introduces an innovative approach to obtaining Gene Expression Associated with a Comparative study between Lepidium sativum extract and hormonal treatment of ovarian inactivity in crossbreed cows with prolificacy in improving reproductive efficiency among different cow

Conflicts Of Interest

The authors declare that there is no conflict of interest.

Ethical Clearance

This work is approved by The Research Ethical Committee.

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