BLOOD PROFILE OF ZINC AND COPPER AT ESTRUS STAGE AND ITS EFFECT ON SUBSEQUENT PREGNANCIES IN BUFFALOES

Rajkumar, R., Singh, S.K., Agarwal, S.K., Mahmood, S. and Shankar, U. 2010. Effect of selective COX- 2 inhibitor on conception rate, progesterone and PGFM profile in buffalo (Bubalus bubalis). J. Appl. Anim. Res, 38: 209–212. To determine the effect of selective COX-2 inhibitor (Meloxicam) on conception rate, progesterone and 13, 14-dihydro-15-keto-prostaglandin F2α(PGFM) profile, buffaloes in estrus were inseminated and divided into two equal groups. Meloxicam was injected @ 0.5 mg/kg body weight in tramuscularly on d 13, 14 and 15 post-insemination to treatment group and normal saline into control group. Conception rate was higher (20%) in the meloxicam treated buffaloes. Serum progesterone concentration did not differ (P>0.05) between the two groups. However, it was higher (P<0.05) on day 18, 21 and 24 post-insemination in pregnant animals indicating maintenance of CL and thus pregnancy. The serum PGFM concentration was (P<0.01) lower on d 15 in meloxicam treated pregnant and non-pregnant buffaloes indicating decrease in PGF2α secretion following administration of a selective COX-2 inhibitor. The results demonstrated beneficial effect of selective COX-2 inhibitor on conception rate in buffalo.

Pregnancy-associated glycoproteins (PAGs) concentrations in water buffaloes (Bubalus bubalis) during gestation and the postpartum period

Simple SummaryToday, the ability to determine the pregnancy status of cows as soon as possible after artificial insemination (AI) has become the most important thing to obtain in an ideal farm. Several efforts have been made to discover biomarkers of early pregnancy but, as of today, without any particular result. Most of the studies were carried out on non-invasive and cheap biological fluids, such as milk. Therefore, in order to identify potential biomarkers of early pregnancy a metabolomic approach on milk of 10 pregnant and 10 non-pregnant buffaloes was used. Milk was recovered in different days before and after the AI, and the data were analyzed retrospectively. The results revealed significant differences between pregnant and non-pregnant buffaloes, as well as in the expression of five metabolites. These data suggest the effectiveness of the metabolomic analysis for the identification of novel potential biomarkers in early prediction of pregnancy in buffaloes after AI, and these findings would give breeders the opportunity to rebreed animals at the next estrus event, saving most of the days as open.This study aimed to identify potential biomarkers for early pregnancy diagnosis in buffaloes subjected to artificial insemination (AI). The study was carried out on 10 pregnant and 10 non-pregnant buffaloes that were synchronized by Ovsynch-Timed Artificial Insemination Program and have undergone the first AI. Furthermore, milk samples were individually collected ten days before AI (the start of the synchronization treatment), on the day of AI, day 7 and 18 after AI, and were analyzed by LC–MS. Statistical analysis was carried out by using Mass Profile Professional (Agilent Technologies, Santa Clara, CA, USA). Metabolomic analysis revealed the presence of several metabolites differentially expressed between pregnant and non-pregnant buffaloes. Among these, a total of five metabolites were identified by comparison with an online database and a standard compound as acetylcarnitine (3-Acetoxy-4-(trimethylammonio)butanoate), arginine-succinic acid hydrate, 5′-O-{[3-({4-[(3aminopropyl)amino]butyl}amino)propyl]carbamoyl}-2′-deoxyadenosine, N-(1-Hydroxy-2-hexadecanyl)pentadecanamide, and N-[2,3-Bis(dodecyloxy)propyl]-L-lysinamide). Interestingly, acetylcarnitine was dominant in milk samples collected from non-pregnant buffaloes. The results obtained from milk metabolic profile and hierarchical clustering analysis revealed significant differences between pregnant and non-pregnant buffaloes, as well as in the metabolite expression. Overall, the findings indicate the potential of milk metabolomics as a powerful tool to identify biomarkers of early pregnancy in buffalo undergoing AI.

Equine chorionic gonadotropin improves the efficacy of a timed artificial insemination protocol in buffalo during the nonbreeding season

This study investigates a novel early pregnancy marker in water buffaloes, focusing on RSAD2 mRNA expression, known to be upregulated by interferon-tau (IFNT) during pregnancy. While RSAD2 is primarily recognized for its antiviral effect, we hypothesized its role as a conceptus-induced component in regulating pregnancy in buffaloes. Given its differential expression compared to other IFNT-induced genes in cows, RSAD2 may serve as a biomarker for early pregnancy detection in buffaloes. RNA, cDNA, and plasma samples were obtained from archived samples collected before insemination (d0) and at d20, d25 and d40 after insemination. Twelve RNA samples, having optimal optical density and concentration, from six pregnant and six non-pregnant buffaloes were selected. The cDNA was analyzed to measure the abundance of RSAD2 mRNA using real-time quantitative PCR (RT-qPCR) and plasma for protein expression analysis using Western blot. The RT-qPCR analysis showed a transcript of RSAD2 increased significantly by 7-fold and 6-fold on d20 and d25, compared to both d0 and d40 in the pregnant group only. At d20, the sensitivity of RSAD2 was 100% and the specificity was 83.3%, and at d25-d both the sensitivity and specificity was 100%, indicating low incidences of misdiagnosing early pregnancy in buffaloes. In the non-pregnant group, RSAD2 expression remained low and did not change after insemination. Western blot analysis revealed an immunoreactive RSAD2 protein band. Densitometry analysis of the RSAD2-specific protein band, based on gray mean value, showed significantly increased expression of RSAD2 at d25 compared to d0 in the pregnant group. In conclusion, these results indicated that RSAD2 expressions at both the mRNA and protein levels show promising potential for detecting pregnancy at d25 post-insemination.

Dietary zinc (Zn) and copper (Cu) supplementation can affect the digestion and absorption of nutrients in pigs, especially of minerals. This study aimed to evaluate the effect of dietary levels of Zn and Cu on their ileal (AID) and total tract (ATTD) digestibility and that of manganese (Mn), calcium (Ca), and phosphorus (P). Six crossbred pigs were surgically equipped with a single-T cannula in their distal ileum. In cross over design, pigs received one of a four corn-soybean meal diets supplemented by two levels of Zn (100 and 500 mg/kg as Zn oxide) and two levels of Cu (40 and 80 mg / kg as Cu sulfate) for each period (7d with 2d for ileal digesta and feces collection). In this study, high level of Zn increased Zn and Mn AID (P&amp;lt; 0.01) but decreased Ca AID (P&amp;lt; 0.05). The high level of Cu improved Cu AID but only when high level of Zn was used (Zn x Cu, P&amp;lt; 0.051). The ATTD of Zn, Cu, Mn, and P were greater in pigs receiving 500 mg/kg of Zn (P&amp;lt; 0.01). The high level of Cu also increased its ATTD (P&amp;lt; 0.01) but reduced that of Ca when low level of Zn was added (Zn x Cu, P&amp;lt; 0.01). The post-ileal digestibility showed a significant absorption of Cu, Mn and Ca (digestibility different from 0). The post-ileal digestibility of Cu, Mn and Ca was greater when high level of Zn was added (P&amp;lt; 0.05). This study confirmed that the AID and ATTD of Cu and Zn could be affected by the dietary level of Zn and Cu in growing pigs. This study also showed that high levels of Cu and Zn reduced ATTD and AID of Ca but increased P ATTD. This last result is contradictory and must be confirmed by the analysis of phytic P.

Early resynchronization of non-pregnant beef cows based in corpus luteum blood flow evaluation 21 days after Timed-AI

Brown-rice originating from local tidal swamp rice is identified as having high iron (Fe) and zinc (Zn) content. This paper aims to elaborate the potentials and prospects of Fe and Zn content of brown rice in tidal swamp local rice, especially from South Kalimantan. This research was carried out at tidal swampland of Belandean Experimental Station, Barito Kuala District South Kalimantan. A total of 71 local varieties of tidal swamp rice originating from the tidal swampland were planted in a 2 m x 5 m plot. The planting system was followed by the South Kalimantan farmers’ traditional system with three times transplanting. The harvested rice was threshed and then milled which was still aleuron. The analysis of Fe and Zn of the brown rice was carried out in the laboratory of The Center for Soil and Agroclimate Research (at present ICALRD) Bogor. The results of the analysis of Fe and Zn levels of rice showed that levels of Fe and Zn of brown rice varied greatly. Fe levels ranged from 11-83 ppm, where Fe levels of local varieties were higher than high yielding varieties. Likewise, levels of Zn with a wide enough interval, which ranged from 20-108 ppm Zn. Among 71 brown rice local varieties, Siam Panangah showed high levels of Fe and Zn, which were 54 ppm Fe and 108 ppm Zn. The brown rice can be used as a source of nutrition Fe and Zn and as the parents in rice breeding with high Fe and Zn content.

Context Antral follicle count (AFC) has been related to bovine fertility and response to reproductive biotechnologies, but its relationship to oestrus expression during timed artificial insemination (TAI) is not known. Aims This study evaluated the relationship between the AFC and oestrus expression in regard to the conception rate in beef cows that received an ovulation synchronisation protocol for TAI. Methods Multiparous Nelore cows up to 40 days postpartum were evaluated by an ultrasound to determine the AFC, and were classified into low (=15 follicles; n = 150), intermediate (=19 and =26 follicles; n = 150) and high count (=30 follicles; n = 150). AFC evaluation was performed on a random day of the oestrous cycle (Day 0). The animals received a conventional TAI protocol based on progesterone and oestradiol benzoate. On Day 8, the animals received cloprostenol, equine chorionic gonadotrophin and oestradiol cypionate, in addition to a painting in the sacrococcygeal region. On Day 10, TAI was performed, and oestrus expression was evaluated and classified as high, low or absent, according to the removal of paint. Pregnancy diagnosis was performed by ultrasound 30 days after TAI. The data were analysed using a generalised linear model and a binary logistic regression model (P &amp;lt; 0.05). Key results Regardless of AFC, a high proportion of cows showed high oestrus expression (low 61.3%; intermediate 67.3% and high 58.7%) at the end of the TAI protocol. The conception rate was influenced by AFC (low 69.3% vs high 56.7%; P = 0.04) and oestrus expression (high oestrus: 67.3% vs no oestrus: 45.4%; P = 0.003). In addition, an interaction (P = 0.008) between AFC and oestrus expression showed that low AFC cows and high oestrus expression had the highest conception rate to TAI. Conclusion AFC and oestrus expression classification influenced the conception rate of Nelore cows in the TAI programme. In addition, the interaction of AFC with oestrus expression revealed that the best conception rate was observed in females with low AFC that expressed high-intensity oestrus. Implications The associated assessment of AFC and oestrus expression represents a strategy to increase the conception rate in TAI programmes in cattle.

Background: The current study aims to measure the effect of oxidative stress on the pregnancy rates of Bulgarian Murrah buffaloes during the breeding and non-breeding season.&#x0D; Methods: The study group consisted of 24 mature buffaloes more than 40 days after parturition. The following parameters were measured: Reactive Oxygen Species (ROS) products, Ascorbate radicals, Malondialdehyde (MDA), Nitric Oxide (NO), Super Oxide Dismutase (SOD), Glutathione peroxidase (GSH-Px), Protein Carbonyl Content (PPC), and total Nitric oxide. The Presynch/Ovsynch protocol was used for estrus synchronization.&#x0D; Results: A statistically significant increase in ROS products were measured in blood serum during the breeding season compared with the non-breeding season. The highest levels measured were in non-pregnant buffaloes during the breeding season. High levels of oxidative stress were registered due to low SOD activity in buffaloes during the breeding season compared to SOD activity during the non-breeding season. The highest SOD activity was observed in non-pregnant buffaloes during the summer season. The lowest GSH-Px levels were observed in non-pregnant buffaloes during both study periods. During the breeding season, concentrations of total NO and PPC were elevated.&#x0D; Conclusion: Comparing the obtained results for oxidative stress and antioxidant activity concerning pregnancy rate depending on the season showed that pregnancy in buffaloes during the breeding season was realized at higher values of NO and SOD. Increased oxidative stress was observed, resulting in a statistically significant increase in serum ROS products, as well as decreased SOD activity in buffaloes during the breeding season.

The role of melatonin as a protective neurohormone against restoring cyclicity in summer anoestrous animals in photoperiod species has gained wider acceptance. This study was designed to uncover the evidence the slow-release melatonin (MLT) has on initiation of ovarian cyclicity and conception rate (CR) in summer anoestrous buffaloes. Thus, buffaloes diagnosed as summer anoestrous (absence of overt signs of oestrus, concurrent rectal examination and radioimmunoassay for serum progesterone at 10 days interval) were grouped as untreated (Group I, sterilized corn oil, n = 8) and treated (Group II, single subcutaneous injection of MLT @18 mg/50 kg bwt in sterilized corn oil, n = 20). Animals treated and detected in oestrus were artificially inseminated (AI) followed by division into Group III (second dose of MLT on 5th day post-AI, n = 8) and Group IV (no melatonin administration, n = 10). Blood samples were collected at 4 days interval for estimation of serum MLT, progesterone and oestrogen using radioimmunoassay kit. Mean oestrous induction rate (OIR), oestrous induction interval (OII), interoestrous interval (IOI) and CR were estimated. Compared to control, concentration of melatonin was significantly (p < 0.05) higher in treated group ranging from 14.34 ± 1.72 to 412.31 ± 14.47 pg/ml whereas other two hormones did not show any concentration difference. Melatonin-administered buffaloes showed significantly (p < 0.05) higher (90%) OIR with OII of 18.06 ± 1.57 days. Results showed improvement in conception rate in buffaloes administered with post-insemination melatonin. It can be concluded from the study that slow-release melatonin supplementation restored cyclicity in summer anoestrous animals resulting in improvement in conception rate in buffaloes.

Conception rates in nulliparous beef heifers can be greatly influenced by their ability to reach puberty at or prior to the initiation of a breeding season. Previous research has analyzed the efficacy of utilizing both long and short-term exposure to a source of progesterone via a controlled internal drug release insert (CIDR) prior to estrous synchronization with timed artificial insemination (TAI) and found improved conception rates. The purpose of this study was to evaluate the effects of feeding melengesterol acetate (MGA) immediately and 10 days before estrous synchronization. Ninety-three crossbred heifers (395.67 ± 5.37 kg) were blocked by BW, genetics, and reproductive tract scores and allotted to 3 treatments. The 3 treatment groups were: 7-day Co-synch + CIDR without presynchronization (CON, n = 31); 2cc PGF2α (Lutalyse) followed by melengesterol acetate feeding for 7 d before the start of the 7-day Co-synch + CIDR program (PRE, n = 31); and 2 mL Lutalyse followed by MGA feeding for 7 d followed 10 d later by the start of the 7-day Co-synch + CIDR program (PRE+10, n = 31). The 7-day Co-synch + CIDR protocol was initiated on d 0 by administering a 2 mL IM injection of GnRH and placing a CIDR into the vagina. The CIDRs were removed 7 days later and accompanied by 2 ml IM injection of PGF2α. An injection of GnRH occurred 60 to 66 hours following PGF2α at TAI with frozen semen from a single bull. Ten days after TAI, heifers were exposed to a bull. Estrotect patches were applied throughout the study to assess estrous behavior. Ovaries were visualized 24 hours post TAI to determine whether ovulation occurred. Pregnancy was determined on days 40, 64, and 109 post TAI via transrectal ultrasonography. Blood was collected via jugular venipuncture (d -19, MGA initiation and termination, d 0, d 7, and d 9) and serum progesterone determined. Performance data were analyzed using the MIXED procedure and conception data were analyzed using the GLIMMIX procedure of SAS. Heifers in the PRE+10 treatment group had higher levels of progesterone (P = 0.04) at d 0 than PRE heifers. At d7 there was a tendency (P = 0.07) for PRE+10 heifers to have greater concentrations of progesterone than PRE, but did not differ by d 9 (TAI, P = 0.36). TAI conception rates in heifers in the PRE treatment group (63%) tended (P = 0.09) to be greater compared with the controls (35%) and not different from PRE+10 (43%), with no differences in season-long pregnancy rates (P &amp;gt; 0.15). Presynchronization with MGA immediately before TAI synchronization can increase conception rates early in the breeding season in beef heifers.

The objectives of the present series of experiments were to develop a timed artificial insemination (TAI) program for dairy heifers and to utilize a TAI program to evaluate the effect of flunixin meglumine (Banamine�; Schering-Plough Animal Health Corp., Atlanta, GA, USA), a prostaglandin H synthase (PGHS)-2 inhibitor, on pregnancy rate and embryo survival. In Experiment 1, 247 heifers were assigned randomly to a prostaglandin F (PGF)/gonadotropin-releasing hormone (GnRH) TAI [i.e., 2 injections of Lutalyse� (25 mg, IM; Pfizer Animal Health Inc., Groton, CT, USA) given 14 days apart in the PM; 60 h after the 2nd Lutalyse injection, heifers received a GnRH injection (Cystorelin�; 100 µg, IM; Merial, Duluth, GA, USA) and were TAI] or a 5-day CIDR/Synch TAI [intravaginal insertion of a CIDR� device (Pfizer Animal Health Inc.) and an injection of GnRH in the AM; 5 days later in the AM, the CIDR insert was removed and Lutalyse was injected, followed by a 2nd injection of Lutalyse� 12 h later; heifers were TAI and injected with GnRH at 72 h after CIDR removal]. Blood samples for progesterone analyses were taken 7 days apart prior to initial PGF or CIDR insertion from the respective groups to determine cycling status. The CIDR/Synch TAI heifers tended to have greater Day 32 (53.1, &gt;46.2%) ultrasound pregnancy rate (USPR) and Day 42 (50.8, &gt;43.7%) rectal palpation (RPPR) than PGF/GnRH heifers (P = 0.10); 4/9 non-cycling heifers of the CIDR/Synch TAI group conceived v. 0/3 of the PGF/GnRH TAI group. In Experiment 2, 176 heifers underwent the CIDR/Synch TAI protocol. However, 2 injections of cloprostenol [500 µg AM/PM; Estrumate�, Schering-Plough Animal Health Corp.) were used to regress the CL at CIDR removal. Heifers at initiation of the CIDR/Synch TAI protocol were assigned randomly to receive injections of Banamine (400 mg IM) at Day 15.5 and at Day 16.0 or no injections (control). Banamine treatment failed to alter either Day 32 USPR (59.6% Banamine v. 59.8% control) or Day 46 RPPR (59.6% Banamine v. 58.6% control). Overall, 4/11 non-cycling heifers conceived. In Experiment 3, 147 heifers underwent a UsedCIDR/Synch TAI protocol utilizing cloprostenol as in Experiment 2. A 5-day used CIDR was employed. Heifers at initiation of the UsedCIDR/Synch TAI protocol were assigned randomly to receive injections of Banamine (400 mg IM) at Day 15.5 and at Day 16.0 or no injections (control). Banamine treatment failed to alter either Day 32 USPR (60.5% Banamine v. 62.0% control) or Day 46 RPPR (59.2% Banamine v. 60.6% control). Pooled overall analyses of pregnancy rates for dairy heifers receiving the CIDR/Synch TAI treatments (n = 451), adjusted for experiments, were 58.3% at Day 32 (USPR) and 57.6% at Day 46 (RPPR). In conclusion, Banamine failed to improve pregnancy rate and/or late embryo survival in dairy heifers, and a CIDR/Synch TAI program is very effective for optimizing the pregnancy rate in dairy heifers. This work was supported by the Florida-Georgia Milk Check-off program.

Resynchronization improves reproductive efficiency of suckled Bos taurus beef cows subjected to spring-summer or autumn-winter breeding season in South Brazil

The aim of this study was to investigate whether there is a correlation between copper (Cu) and zinc (Zn) levels in children and their parents, considering their nutritional habits. Cu and Zn concentrations were measured by flame atomic absorption spectrophotometry in the serum of 66 healthy children, aged 3-14 years, and their parents, residing in a region of Greece (Thrace). Cu levels were higher in mothers than those in fathers, but they were lower in both parents than those in children. They also tended to decrease with age in both parents and children, whereas Zn levels significantly increased with age in children. There was a positive correlation between children's and mothers' Zn levels, as well as children's and both parents' Cu levels. Children used to eat meat, fish, vegetables, and legumes as frequently as their parents, but they were consuming more eggs, milk, and fruits than the latest. Regarding parents' diet, higher Zn levels were depended on the consumption of meat and milk, whereas higher Cu levels were depended on the consumption of milk. Consequently, children's Cu and Zn levels are related to their parents' levels, which can be influenced by their nutritional habits.