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The aims of this study were to assess the efficacy of alphacypermethrin (ACYP) on pediculosis due to Haematopinus tuberculatus and to evaluate the influence of the treatment on productive and reproductive performance in buffaloes (Bubalus bubalis) reared in an intensive system. The trial was performed on 56 pluriparous buffaloes at 86.8 ± 8.1 d in milk. The animals underwent individual louse count and were divided into 2 homogenous groups according to louse count, age, number of lactations, days in milk, live BW, BCS, pregnancy status, and milk yield. Group A (n = 28) was treated by a pour-on formulation of ACYP, and Group S (n = 28) was treated by pour-on saline solution. Individual louse counts were performed weekly on 10 buffaloes in each group. Feed intake was recorded daily and the total mixed ration, individual ingredients, and orts were analyzed to calculate DM ingestion. Individual milk yield was recorded daily and milk samples were analyzed at the beginning of the trial, after 4 wk, and at the end of the trial to assess milk composition. Individual BCS was also evaluated simultaneously. Finally, the animals underwent synchronization of ovulation starting 4 wk after treatment and the pregnancy rate and the calving-conception interval were evaluated. Data were analyzed by the Mann-Whitney test and ANOVA for repeated measures. The infestation was constant in Group S, whereas no lice were present in Group A throughout the study. Daily DMI was similar in the 2 groups (16.7 ± 0.4 vs. 16.3 ± 0.3 kg/d in Group A vs. Group S, respectively), although buffaloes in Group A showed higher (P < 0.05) BCS score at the end of the trial (7.39 ± 0.1 vs. 7.14 ± 0.1 in Group A vs. Group S, respectively). The average milk yield/buffalo was higher (P < 0.05) in Group A compared to Group S (10.58 ± 0.1 vs. 10.39 ± 0.1 kg in Group A vs. Group S, respectively) and this was mainly due to the higher milk production recorded in buffaloes at less than 75 d in milk (11.81 ± 0.1 vs. 11.45 ± 0.1 kg in Group A vs. Group S, respectively). Despite of a similar fertility rate (90.5 vs. 80.9% in Group A vs. Group S, respectively), a lower (P < 0.05) calving-conception interval was recorded in Group A compared to Group S (118 ± 16 vs. 177 ± 16 d in Group A vs. Group S, respectively). In addition to the pour-on treatment against pediculosis, productive and reproductive performance were also improved. This represents a significant improvement in dairy buffalo herd management.

The aim of this study was to evaluate the efficacy of tulathromycin in the control of respiratory diseases (BRD) in buffalo calves. The trial was performed in one buffalo farm located in the South of Italy where more than 2,000 animals are bred. An outbreak of BRD was considered when the animals showed three of the following criteria: rectal temperature >39.8°C, depression score >2 on a scale of 0 (alert) to 4 (soporous), respiratory rate >60 ⁄ min), dyspnoea (abnormal lung sounds) and cough or nasal discharge. A nasal swab was performed for microbiological investigations. After the results of the latter animals were treated with a standard therapeutic regimen on D 0, which consists of 2.5 mg/kg/live weight of tulathromycin (Draxxin®, Zoetis SRL, Italy) in a single administration. Daily clinical examinations were performed on treated calves for fourteen consecutive days to evaluate: A) depression score and general health status with a score from 0 (normal) to 4 (general conditions severely impaired); B) respiratory distress with a score from 0 (no alterations) to 3 (respiratory rate >80⁄min, abdominal breath and nasal discharge) and C) rectal temperature with a score from 0 (normal; 38.8-39°C) to 3 (> 40.3°C). Data were analyzed by analysis of variance. A total of 81 calves (53 females and 28 males) were included in the study. Bacteriological cultures revealed the presence of Pasteurella M. and Moraxella spp. in 53.1 and 38.3% of the samples, respectively. The average A, B and C scores were 2.24±0.1, 1.85±0.1 and 1.78±0.1 on day 0. Treatment was effective in solving the BRD in four days (Scores A, B and C = 0) in 79/81 calves (97.5%), and, consequently, 2/81 (2.5%) mortality rate was recorded. In conclusion, treatment with tulathromycin is highly effective in the control of the BRD in buffalo calves.

The trial was performed on 18 multiparous Mediterranean buffaloes, pre-synchronized by a double injection of 0.524 mg of PGF2α. Animals were divided into two groups (A and B), that underwent two superovulation (SO) treatments, starting 8 days after estrus (day 0): Group A (n=9) received a 4-days double daily decreasing dosage of 400 mg NIH-FSH-P1 (Folltropin®, Bio98, Italy). Group B (n=9) received the same treatment as Group A for the first 2 days, and a single injection of 1000 IU of eCG (Ciclogonina®, Fortdodge, Italy). On day 10 all buffaloes received a double injection of 0.524 mg of PGF2α to induce luteolysis. Ultrasound follicular development was assessed daily on each buffalo from the start of the SO treatments until 48 hours after estrus detection to record the total number of the follicles, grouped into 3 categories according to their diameter: small (between 0.20 and 0.50 cm), medium (between 0.51 and 1.00 cm) and large (more than 1.01 cm). The statistical analysis was carried out by using the chi-square test and the analysis of variance. Buffaloes in Group B showed a higher (P

Italy can be considered a leading country for both genetic improvement and breeding techniques in buffalo species. However, few information are actually available on physiology of this species and particularly those for high producing dairy buffaloes maintained in intensive and semi-intensive breeding. Therefore, it is essential to carry out new studies aimed to increase metabolism knowledge in this species. The purposes of this doctoral thesis were to evaluate: 1) the results of the application of the out of season breeding mating technique and the metabolic status of buffaloes undergone milk recording; 2) the metabolic changes that occur throughout the lactation and in different climate conditions; 3) the possibility of using some parameters assessed in the milk as risk indexes for metabolic diseases; 4) the influence of milk production and some metabolites on pregnancy. Two trials were carried out in the experimental design. The main aim of Experiment 1 was to verify milk production and Beta-Hydroxybutyrate (BHBA) levels in primiparous and pluriparous buffaloes undergone milk recording. Experiment 2 was carried out simultaneously in 8 commercial buffalo farms located in the area of buffalo mozzarella cheese DOP production. The feedstuffs utilized for animal feeding were collected in each farm and analyzed for chemical composition. Milk yield was recorded monthly and two milk samples (during morning and afternoon milking) were collected to evaluate: fat, protein, lactose, urea, true protein, casein, CF, H-index, cryoscopic index, acetone, BHBA and fat characteristics (satured and unsatured fats). Furthermore, 40 buffaloes with different days in milk (5-50 days; 50-120 days; 120-200 daysand 200-270 days) were selected in each farms to analyze metabolic profile and its relationship with metabolites in milk. Therefore, these buffaloes underwent blood sampling two months apart when milk recordings were carried out to analyze the metabolic profile (see below). Throughout the experimental, period 3 milk tank sampling were performed in 6 farms to perform chemical analysis and cheese yield. Finally, the influence of metabolic status on reproductive performance was assessed in each farm, mainly looking at oocyte and embryo development. Therefore, a representative number of buffaloes in each farm underwent synchronization of ovulation and artificial insemination. The results of Experiment 1 demonstrated that the out of season breeding mating technique is correctly applied in primiparous buffaloes, while a higher seasonal pattern can be observed in pluriparous counterparts. Furthermore, BHBA levels were higher at both the beginning and the end of lactation, compared to the mid lactation period. The analysis of urea levels in milk support the evidence that protein requirements were not satisfied or exceeded the requirements in about 50% of cases. In Experiment 2, it was observed that the number of days in milk and the season influence energy metabolism through a lower energy intake, higher metabolic strain (for both the end of lactation and cold periods) and higher milk yield. These buffaloes underwent an excess of fat mobilization, that was the main reason for milk qualitative characteristics worsening and lower yield in mozzarella cheese. Furthermore, the lower dry matter intake and the lipomobilization, recorded at the beginning of lactation caused a change in fat characteristics, as demonstrated by the evidence that buffaloes at lower than 50 days in milk showed a significantly (P

The aim of this study was to evaluate water balance and nitrogen excretion in Mediterranean buffaloes. Sixteen subjects were divided into 4 groups (n=4/group), according to their days in milk (DIM): Group L50 (DIM=50±8); Group L125 (DIM=125±55); Group L225 (DIM=225±26); Group NL (non-lactating). The study comprised a 14-day adaptation to diets and 14-day data collection during which feed intake and milk production were recorded daily. Measurements for individual buffaloes were made for water intake, diet, orts, milk, faeces and urine during the final 3 days. The analyses were performed according to OAOC. Lactating buffaloes had greater (P

The aim of this work was to evaluate whether L-carnitine supplementation during IVC improves blastocyst development and cryotolerance of in vitro produced (IVP) buffalo embryos. Abattoir-derived cumulus-oocytes complexes (n=410, over 5 replicates) were matured and fertilized in vitro according to standard procedures (Gasparrini B et al. 2006 Theriogenology 65 (2), 275-287). On day 1 (Day 0 = IVF), zygotes were cultured in SOF supplemented with 8 mg/ml BSA, in the absence (control, n=165) or presence of L-carnitine (n=170) at a concentration (0.25 Mm) selected after a preliminary dose response trial. In vitro culture was carried out at 38.5°C under 5% CO2, 7% O2, and 88% N2. Cleavage and blastocyst rates (in relation to the cleaved embryos) were evaluated on Day 5 and 7, respectively. The blastocysts were vitrified by cryotop in 16.5% ethylene glycol, 16.5% DMSO and 0.5M sucrose and the survival rate, based on morphological criteria, was assessed after 24 h culture. Data were analyzed by Chi square test. Cleavage (81.3% vs 82.1%, in the control and carnitine groups) and blastocyst production (40.0% vs 47.6%, in the control and carnitine groups) were not affected by the treatment. The percentages of fast developing embryos (expanded and hatched blastocysts), i.e. those of better quality, were 17.0 and 23.5%, respectively. Interestingly, the embryos cultured with L-carnitine showed higher survival rates after 24 h culture (78.7% and 96.4%, in the control and carnitine groups, respectively; P

The aim of the present study was to compare two synchronization protocols for timed artificial insemination (TAI) in acyclic pluriparous buffalo cows during the non breeding season. Two experiments were conducted to evaluate the ovarian follicular response and pregnancy rate. The cyclic status was evaluated by two transrectal ultrasonography performed at Day -11 and Day 0. Buffaloes that in both investigations did not show the presence of a corpus luteum (CL) were classified as acyclic. Acyclic pluriparous buffaloes (n=34) were randomly assigned to Group 1 (G1) and Group 2 (G2), homogeneous for Days in Milk (81±27 vs 83±13, respectively in G1 and G2). In G1 (n=17) buffaloes received 12μg of buserelin acetate i.m. (GnRH) on the first day of the synchronization protocol (Day 0), 0.524 mg of cloprostenol (Pgf2α) on Day 7 and 12μg of buserelin acetate i.m. on day 9 (Ovsynch-TAI). In G2 (n=17) buffaloes received a progesterone-releasing intravaginal device containing 1.55 g of progesterone (P4) and 12μg of buserelin acetate i.m. (GnRH) on Day 0. On Day 8 the P4 device was removed and 0.524 mg of cloprostenol (PGF2α) + 500UI of PMSG i.m. were administered. Finally, 12μg of buserelin acetate i.m. were given on Day 10. Ten animals (5/group) underwent transrectal ultrasonography of the ovaries daily, from Day 0 to Day 11 in G1 and from Day 0 to Day 12 in G2, to determine the presence and diameter of the follicles, the dominant follicle (DF) diameter and the ovulation rate. Subsequently, fixed TAI was performed 20 hours after the last GnRH in all buffaloes. Ultrasonography was carried out 25 and 45 days after TAI to evaluate pregnancy rate and the incidence of late embryonic mortality (LEM). No differences were observed between G1 and G2 in the following parameters: DF diameter (mm) on Day 0 (9.5±1.9 vs 8.8±4.3), DF diameter (mm) on Day of Pgf2α administration (11.5±2.3 vs 10.6±1.6), DF growth rate (mm) between PGF2α and last GnRH administration (1.6±0.3 vs 1.8±1.7), DF maximum diameter (13.2±1.9 vs 13.0±2.3 mm) and ovulation rate (80% vs 80%). However, in G2 pregnancy rate increased at 25 (29.4% vs 58.8%, respectively in G1and G2; P= 0.08) and 45 (23.5% vs 58.8%, respectively in G1and G2; P

Primiparous buffaloes were tested in two periods of the year characterized, by either low or high reproductive efficiency. They were subjected to two protocols for synchronization of ovulation: (i) Ovsynch (OV) and (ii) progesterone based (P4) treatment. After calving, the animals underwent a series of four cycles of re-synchronization protocols. The season did not affect pregnancy rates when the results of the two treatments were pooled together with regard to the first synchronization protocol, followed by AI. Pregnancy rates were similar during the low breeding season (50.3% vs. 57.4% in OV and P4, respectively), but different during the high breeding season (50.4% vs. 67.7% in OV and P4, respectively; p = 0.000). Logistic regression confirmed a significant effect of treatment and season interaction on pregnancy (p = 0.003). Following re-synchronization, a treatment by season interaction was detected during the low breeding season (odds ratio = 2.233), in favor of P4. Finally, a survival analysis showed a better response of animals subjected to P4 treatment from the second AI onward. In conclusion, the pooled data of pregnancy rates from both treatments between seasons are not different following AIs. Better results, though, were obtained from the implementation of P4 treatment, and are recorded in a season-fashioned mode when the comparison is made following first or cumulative AIs.

In the present study, we aimed at assessing the influence of breed and feeding system on the bovine milk profile of betaines and carnitines and milk capacity in counteracting the inflammatory endothelial cell (EC) damage induced by interleukin (IL)-6. In the first experimental design, two breeds were chosen (Holstein vs. Modicana) to investigate the biomolecule content and antioxidant capacity in milk and dairy products. In the second experimental design, two feeding systems (pasture vs. total mixed ratio) were tested only in Holstein to evaluate the possible effect on the functional profile of milk and dairy products. Finally, the bulk milk from the two experimental designs was used to evaluate the efficacy of preventing IL-6-induced endothelial inflammatory damage. Results showed that Modicana milk and whey had higher biomolecule content and antioxidant activity compared to Holstein milk (p < 0.01). Milk from Holstein fed TMR showed higher concentration of γ-butyrobetaine, δ-valerobetaine (p < 0.01), and l-carnitine (p < 0.05). Similarly, whey from Holstein fed TMR also showed higher content of δ-valerobetaine, glycine betaine, l-carnitine, and acetyl-l-carnitine (p < 0.01) compared to the Holstein fed pasture. Conversely, the antioxidant activity of milk and dairy products was not affected by the feeding system. In ECs, all milk samples reduced the IL-6-induced cytokine release, as well as the accumulation of reactive oxygen species (ROS) and the induction of cell death, with the most robust effect elicited by Modicana milk (p < 0.01). Overall, Modicana milk showed a higher content of biomolecules and antioxidant activity compared to Holstein, suggesting that the breed, more than the feeding system, can positively affect the health-promoting profile of dairy cattle milk.

The aim of the study was to evaluate whether resveratrol supplementation of bovine culture medium improves in vitro blastocyst development, embryo cryotolerance and cell numbers. Abattoir-derived oocytes were matured and fertilized in vitro according to standard procedure. Twenty hours after IVF, zygotes were cultured in SOF medium, supplemented with 0 (control, n=439), 0.25μM (n=422), 0.5μM (n=447) and 1μM resveratrol (n=416). On Day 7 (IVF=Day 0) blastocysts were vitrified by cryotop in 16.5% ethylene glycol, 16.5% dimethyl sulfoxide and 0.5M sucrose. Development rate, i.e. the percentage of embryos resuming development to reach a more advanced stage, and hatching rate were evaluated after 24 and 48h culture. Blastocysts cultured with (0.5μM) and without resveratrol underwent differential staining to count inner cell mass (ICM) and trophectoderm (TE) cells. Resveratrol during culture did not increase blastocyst yields (57.1, 57.7, 59.2 and 46.6%, respectively in 0, 0.25, 0.5 and 1μM resveratrol). However, 0.5μM resveratrol improved embryo cryotolerance compared to the control, as indicated by higher development rates (67.3% vs 50.3%, respectively; P

The aim of this study was to evaluate whether supplementation of bovine culture medium with the natural antioxidant L-ergothioneine (LE), improves in vitro blastocyst development and quality, assessed as resistance to cryopreservation, total cells number, cellular differentiation, and apoptosis index. Abattoir-derived oocytes were matured and fertilized in vitro according to standard procedure. Twenty hours after IVF, presumptive zygotes were cultured in synthetic oviduct fluid with 0, 0.05 mM, 0.1 mM, 0.5 mM, and 1 mM of LE (experiment 1) at 39 °C under humidified air with 5% CO2, 7% O2, and 88% N2. On the basis of the results of this dose-response trial, the range of concentrations to test was reduced in experiment 2, in which presumptive zygotes were cultured with 0, 0.05 mM, and 0.1 mM of LE. On Day 7, embryo yields were assessed, and the blastocysts (BL) were vitrified by Cryotop method in 16.5% ethylene glycol, 16.5% DMSO and 0.5 M sucrose. Finally, BL produced on Day 8 in the absence (control) and presence of 0.1 mM LE were used for transferase-mediated dUTP nick end labeling and differential staining to evaluate, respectively the apoptotic rate and the allocation of cells into inner cell mass (ICM) and trophectoderm lineages (experiment 3). Despite similar blastocyst yields, supplementation of culture medium with 0.1 mM LE improved the cryotolerance of in vitro-produced (IVP) embryos compared to the control group, as indicated by higher (P < 0.05) hatching rates recorded after 48-hour post-warming culture (48.5%, 50.0%, and 63.8%, respectively with 0, 0.05, and 0.1 mM LE). Interestingly, when embryos were cultured in the presence of 0.1 mM LE, the percentage of BL with the most physiological ICM:total cells ratio (20%-40%) increased (85.1 vs. 66.0%, P < 0.05), confirming a beneficial effect on embryo quality. Furthermore, 0.1 mM LE decreased (P < 0.01) both the average number (4.3 ± 0.2 vs. 9.1 ± 0.3) and the proportion (3.6 ± 0.3 vs. 8.1 ± 0.5) of apoptotic cells in BL compared to the control. In conclusion, the enrichment of bovine culture medium with 0.1 mM LE improves embryo quality, as indicated by the improved cryotolerance, the lower apoptotic rate, and the higher percentage of BL with the most physiological ICM:total cells ratio.

The aim of this study was to quantify nitrogen (N) and phosphorus (P) utilisation and excretion in intensive dairy buffaloes. Italian Mediterranean buffaloes were divided into 4 groups (n=6/group) as follows: Groups M50, M125 and M225 at 50, 125 and 225 days of lactation, respectively, and Group NL which was non-lactating. Lactating buffaloes had greater (P