Validation of ELISA kits for determination of Inhibin-A and Estradiol-17-beta concentrations in Buffalo plasma

The aim was to evaluate the suitability of two commercial ELISA kits for human serum or plasma, to measure Inhibin-A (In-A) and Estradiol-17-beta (E2) concentrations in buffalo plasma. Blood samples were obtained by jugular venipuncture from buffalo heifers and cows, and plasma samples were stored at – 20°C until assays. Precision of the methods was evaluated by the intra- and inter-assay coefficients of variation (CVs) of buffalo plasma sample replicates, at different concentrations. Accuracy was evaluated calculating the recovery rates of different proportions of the highest standard added to a buffalo plasma sample at low concentration (observed/expected values x 100). Linearity was evaluated by serially diluting one buffalo plasma sample at high concentration with the assay buffer and calculating by regression analysis the parallelism of the resulting line with the standard line. Intra-assay CVs were 11% and 15.1% for In-A and 1.8% and 3.3% for E2. Inter-assay CVs were 13.9% and 7.4% for In-A and E2, respectively. Mean recovery rate was 97.9% and 98.5% for In-A and E2, respectively. Dilution tests gave good parallelism between the lines obtained and the standard lines. It is concluded that the kits tested are suitable and reliable for buffalo plasma samples

Follicular development, plasma Inhibin-A and Estradiol-17-beta concentrations in Buffalo cows during different treatment schedules for MOET programs

Buffalo cows were submitted to three superovulatory treatments. T1 (n = 7): PRID for 10 days (d0-d9) plus decreasing doses of 500 IU FSH/LH (12 h-intervals d7‑d10); T2 (n = 8): PRID for 11 d (d0-d10) plus 2000 IU PMSG at d7; T3 (n = 9): PRID for 11 d plus 2000 IU PMSG at d7 and decreasing doses of 175 IU FSH/LH (12 h-intervals d10‑ d11). Overall plasma inhibin‑A (In-A) concentrations correlated with large follicles (LF, diameter >6mm, R=0.83, P10 mm at d12- 13 (T1=5.0+/-1.4, T2=1.2+/-0.9, T3=8.3+/-2.3). In-A concentrations significantly rised at d11-13 of T1 and T3. In-A seems a good indicator of the follicular development during superovulation in buffalo cows, while E2 is not. Furthermore T3 was followed by better ovarian follicular responses

LH peak and ovulation in buffalo cows treated for oestrus synchronisation using two different hormonal schedule

Since oestrus behaviour in buffalo is frequently scarcely clear and the oestrus signs, even when showed, are not reliable, ovulation can not be predicted. Furthermore high individual variability of the oestrus signs – LH peak interval was observed both in spontaneous and hormonal induced oestrus (Moioli et al., 1998; Barile et al, 1998)

Effects of two different blends of naturally mycotoxin-contaminated maize meal on growth and metabolic profile in replacement heifers

The aim of this trial was to assess the effects of the administration of different combinations of mycotoxins in naturally contaminated maize grains on dairy heifer growth, blood measurements and puberty onset. A total of 35 Friesian female heifers were randomly allotted to three experimental groups from 18-21 to 42-45 weeks of age. During the 24-week experimental period (EP), heifers were fed the same diet, but with maize meal derived from three differently contaminated lots: very low contamination, as control (C); medium-low aflatoxin-contaminated (A); and mixed aflatoxin-fumonisin contaminated (A-F). At the end of the EP, they returned to a common diet without contaminated maize, and they were monitored for an additional period of 12 weeks (post-experimental period, PEP). BW, wither height, hip height, body length and heart girth were measured every 4 weeks from the beginning of EP to the end of PEP. At the same time, body condition score was evaluated and blood samples were taken from the jugular vein to be analysed for haematological, serum protein and metabolic profiles. Age at puberty was assessed by measuring weekly plasma progesterone levels from 40 to 52 weeks of age. Body growth measurements were processed both by ANOVA of average daily gain of EP and PEP separately, and by the analysis of growth curve parameters. Haematological, serum protein and metabolic profile were evaluated using a mixed model, taking into account the repeated measurements in time on each animal. Heifers' growth was delayed both in A and A-F groups during EP, as evidenced by the different linear coefficients of the BW growth curve in the three groups. Differently contaminated diets did not affect the haematological profile, so that it can be concluded that these levels of mycotoxin contamination do not determine any specific effect on haematopoiesis and immunity in growing heifers. The main blood marker of mycotoxin chronic toxicity was the γ-glutamyl transferase activity level in plasma, which appeared to be altered even after the removal of mycotoxins. During EP, plasma glucose was lower in the groups fed contaminated diet compared with C. The joint actions of an altered nutritional status and a long-lasting liver damage were probably the causes of the delay in puberty attainment in A and, particularly, in the A-F group. The results from this trial evidenced that a chronic aflatoxin-fumonisin contamination in diets of dairy heifers can determine an important delay in the reproductive career of these animals

Validation of ELISA kits for determination of Inhibin-A and Estradiol-17-beta concentrations in Buffalo plasma

The aim was to evaluate the suitability of two commercial ELISA kits for human serum or plasma, to measure Inhibin-A (In-A) and Estradiol-17-beta (E2) concentrations in buffalo plasma. Blood samples were obtained by jugular venipuncture from buffalo heifers and cows, and plasma samples were stored at – 20°C until assays. Precision of the methods was evaluated by the intra- and inter-assay coefficients of variation (CVs) of buffalo plasma sample replicates, at different concentrations. Accuracy was evaluated calculating the recovery rates of different proportions of the highest standard added to a buffalo plasma sample at low concentration (observed/expected values x 100). Linearity was evaluated by serially diluting one buffalo plasma sample at high concentration with the assay buffer and calculating by regression analysis the parallelism of the resulting line with the standard line. Intra-assay CVs were 11% and 15.1% for In-A and 1.8% and 3.3% for E2. Inter-assay CVs were 13.9% and 7.4% for In-A and E2, respectively. Mean recovery rate was 97.9% and 98.5% for In-A and E2, respectively. Dilution tests gave good parallelism between the lines obtained and the standard lines. It is concluded that the kits tested are suitable and reliable for buffalo plasma samples

Leptin level in plasma of lactating buffaloes fed two diets with different energy and protein concentrations

Leptin, a protein mainly secreted from the white adipocytes, has been shown to contribute to the regulation of energy metabolism, feeding behaviour and whole body energy balance. Moreover, leptin gene activity and leptin secretion are correlated with body adiposity and changes in food intake. Furthermore, leptin could also modulate endocrine response to changes in nutritional status and/or tissue sensitivity to hormones (Houseknecht et al., 1998; Romsos, 1998). Several factors are known to influence plasma leptin in rodents and humans: particularly it increases by body fatness, insulin, glucocorticoids, estrogens and decreases by food deprivation (Saladin et al., 1995; Ahima et al., 1996; Shimizu et al., 1997). These ones and several other observations have led to the hypothesis that leptin is a signal arising from adipose tissue, linked to the level of fat reserves and/or the nutritional status. This signal directly influences the central nervous system and peripheral organs, resulting in a better adaptation of body metabolism and physiological functions to the availability of metabolic energy..........

Input-output cost of agronomic production of crops under rotate growing condition for applying to Mediterranean Italian buffaloes

Forage availability with low cost of production is the main farmer requisite for obtaining a competitive husbandry dairy product in the market. Cropping system for silage consumption differently impact the cost of Milk Feeding Unit (MFU) and sustainable agronomic cultivation of herbage production. The experiment aimed to assess the analytical cost of MFU per hectare in four forage cropping system models based on two crops per year bred under irrigated and rainfed condition in a Mediterranean site with intensive buffaloes breeding pressure

Effect of different FSH/LH ratios on superovulatory response and embryo yield in goats

Previous studies carried out in sheep (Chupin et al., 1985) and in goats (Nowshari et al., 1995; Martemucci et al., 1996) have underlined the importance of both FSH and LH to induce a good superovulatory response, but the results on the LH amount necessary to give a higher ovulatory response and embryo production are rather contradictory. The aim of this study was to evaluate, in goats, the effect of 2 different FSH/LH ratios (1:1 vs 2:1) kept constant during the treatment, on ovulatory response and embryo production. Moreover, according to the FSH/LH ratio=2:1,........