Early Prediction of Corpus Luteum Functionality Using an Imaging Software

The present study aimed to assess the applicability of luteal blood flow data acquired through the use of color Doppler ultrasonography and a post-processing analysis tool (ImageJ) for predicting pregnancy in buffaloes (Bubalus bubalis). The experiment was carried out on 59 multiparous Italian Mediterranean buffaloes that underwent synchronization of estrus and fixed-time artificial insemination (TAI). Corpus luteum features (size: CLS and blood flow: BFA) were taken from Day 5 to 10 after TAI and retrospectively measured with ImageJ. In the same period, blood samples were taken to assess progesterone (P4) concentrations. Pregnancy diagnosis was carried out on Day 45 by ultrasound and confirmed on Day 70 post-TAI. Differences in CLS, BFA, and P4 concentrations from Day 5 to 10 after TAI measured between groups were analyzed by ANOVA repeated measures as were differences within each day of measuring. Buffaloes that established a pregnancy (n = 29; 55%) had larger CLS (2.2 ± 0.1 vs. 1.9 ± 0.1 cm2; P < 0.01), higher BFA (0.6 ± 0.0 vs. 0.4 ± 0.0 cm2; P < 0.01), and higher P4 blood level (1.8 ± 0.1 vs. 1.4 ± 0.1; P < 0.01) during Day 5–10 as compared to not-pregnant buffaloes (n = 22). Throughout the entire period, the first feature that changed between groups was P4 blood concentration at Day 7 (1.7 ± 0.1 vs. 1.2 ± 0.1; P < 0.05) followed by BFA at Day 8 (0.6 ± 0.0 vs. 0.5 ± 0.0; P < 0.05), respectively, in pregnant and not-pregnant animals. The ROC analyses indicated that P4 was able to predict pregnancy since Day 5 (P < 0.05) although a more reliable result could be obtained from Day 8 (P < 0.01). At Day 10, it was possible to set a cutoff value for every parameter taken into account. The logistic regression analysis showed that pregnancy was positively influenced by P4 concentration (odds ratio 534.127; P < 0.01) and BFA (odds ratio 744.893; P < 0.01). In conclusion, the use of color Doppler ultrasonography, together with ImageJ, identified different patterns of BFA between pregnant and not-pregnant buffaloes starting from Day 8 post-TAI

Relationship among Milk Conductivity, Production Traits, and Somatic Cell Score in the Italian Mediterranean Buffalo

The measurement of milk electrical conductivity (EC) is a relatively simple and inexpensive technique that has been evaluated as a routine method for the diagnosis of mastitis in dairy farms. The aim of this study was to obtain further knowledge on relationships between EC, production traits and somatic cell count (SCC) in Italian Mediterranean Buffalo. The original dataset included 5411 records collected from 808 buffalo cows. Two mixed models were used to evaluate both the effect of EC on MY, PP and FP and EC at test-day, and the effect of EC on somatic cell score (SCS) by using five different parameters (EC_param), namely: EC collected at the official milk recording test day (EC_day0), EC collected 3 days before official milk recording (EC_day3), and three statistics calculated from EC collected 1, 3 and 5 days before each test-day, respectively. All effects included in the model were significant for all traits, with the only exception of the effect of EC nested within parity for FP. The relationship between EC and SCS was always positive, but of different magnitude according to the parity. The regression of EC on SCS at test-day using different EC parameters was always significant except when the regression parameter was the slope obtained from a linear regression of EC collected over the 5-day period. Moreover, in order to evaluate how well the different models fit the data, three parameters were used: the Average Information Criteria (AIC), the marginal R2 and the conditional R2. According to AIC and to both the Marginal and Conditional R2, the best results were obtained when the regression parameter was the mean EC estimated over the 5-day period

Efficacy and Safety of Neem Oil for the Topical Treatment of Bloodsucking Lice Linognathus stenopsis in Goats under Field Conditions

: The aim of the present study was to evaluate the efficacy and safety of neem oil on caprine pediculosis and on kids' growth performances. The neem (Azadirachta indica) belongs to the Meliaceae family, and in Eastern countries it is mainly considered for the insecticidal activities of the kernel oil. The neem seeds contain bioactive principles, such as azadirachtin A, salannin, nimbin, and nimbolide. The trial was carried out on 24 kids, 120 days old, maintained in open yards. Animals were divided in 4 homogeneous groups (n = 6 animals/group) based on age, louse count, body condition score (BCS) and live body weight: Control Group (C, saline NaCl, 0.9%), Neem Group 1 (NO-100, 100 mL of neem oil per 10 kg), Neem Group 2 (NO-200, 200 mL/10 kg), Neem Group 3 (NO-300, 300 mL/10 kg). The treatments were performed by spraying the insecticide on the goat's body. The study lasted 56 days, and weekly, the kids underwent louse count, BCS and body weight determination, and FAMACHA score. Data were analyzed by ANOVA for repeated measures. The species of lice identified was Linognathus stenopsis. Kids belonging to NO-200 and NO-300 showed a stronger reduction of louse count throughout the study (>95%). The daily weight gain recorded was significantly higher (p < 0.05) in NO-300 than C. No differences were found for BCS and FAMACHA scores. The results of this trial showed that the administration of neem oil to control caprine pediculosis caused by sucking lice represents an alternative to synthetic compounds

Breed and Feeding System Impact the Bioactive Anti-Inflammatory Properties of Bovine Milk

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

Investigation of Climate Effects on the Physiological Parameters of Dairy Livestock (Cow vs. Buffalo)

Nowadays climate change is affecting the planet’s biodiversity, and livestock practices must adapt themselves to improve production without affecting animal welfare. This work investigates the influence that some climatic parameters such as Environment Temperature, Relative Humidity, Thermal excursion and Temperature–Humidity Index (THI), can have on milk quantity and quality in two different dairy species (buffaloes and cows) raised on the same farm. A further aim was to understand if THI threshold used for cows could also be used for buffaloes. The climatic parameters were recorded daily through a meteorological station located inside the farm. Milk quantity (converted into ECM) and quality (Fat Percentage—FP; Protein Percentage—PP; Somatic Cell Count—SCC) were measured. Data were analyzed with Spearman’s correlation index, separately for buffaloes and cows. The results indicate a greater sensitivity of cows to heat stress and a strong negative correlation of the ECM with meteorological data (p < 0.01). The results of this study may stimulate the use of integrated technologies (sensors, software) in the dairy sector, since the IoT (sensors, software) helps to enhance animal well-being and to optimize process costs, with a precision livestock farming approach

Influence of Days after Calving and Thermal Stress on the Efficacy of a Progesterone-Based Treatment in Acyclic Italian Mediterranean Buffalo

The aim of this study was to evaluate the efficacy of a progesterone-based treatment on anoestrus in buffaloes. Primiparous acyclic buffaloes (n = 276), were divided into three classes according to their days in milk (DIM): from 50 to 90 (Class I; n = 86), from 91 to 150 (Class II; n = 102) and from 150 to 200 (Class III; n = 88). Animals were synchronized using P4 vaginal implants, followed by timed artificial insemination (TAI). They were then allowed to enter into a larger group of buffaloes for natural mating 15 days after AI was performed, and pregnancy status was monitored from then on at 15-day intervals. Finally, the temperature–humidity index (THI) was calculated. Statistical analysis was performed by ANOVA by means and both multiple and linear regression. The total pregnancy rate (PR) was 87.7%, with no differences among DIM classes (88.0, 92.4, and 80.0% in Classes I, II, and III, respectively). However, the PR at TAI tended to be higher (p = 0.07) in buffaloes in Class II. The follicle (FL) area in Class II buffaloes was larger (p &lt; 0.01) than that of the other classes. No influence of the THI on the total PR was recorded. The pregnancy outcome at TAI was affected by the FL area (odds ratio = 2.237; p &lt; 0.05) and body condition score (BCS) (odds ratio = 1.256; p &lt; 0.05). In conclusion, treatment with vaginal P4 optimizes pregnancy rates in anoestrus buffaloes, particularly when the animals are in mid-lactation and show an optimal BCS. Furthermore, the THI does not seem to affect the efficiency of the progesterone treatment

Relationship among production traits, somatic cell score and temperature–humidity index in the Italian Mediterranean Buffalo

The temperature–humidity index (THI) has been commonly used to analyse heat stress in dairy cattle, but little is known about its effects on buffaloes. In this study, daily milk yield (MY), fat percentage (FP), protein percentage (PP) and somatic cell count (SCC) data from 808 buffalo cows plus environmental temperature and relative humidity were used to investigate the consequence of heat stress. Two mixed models were used to evaluate the impact of THI on MY, FP, PP and log transformed SCC (SCS). The effect of THI was significant for PP, FP and SCS, whereas its interaction with parity was statistically significant for PP and SCS. The relationship between PP and FP and THI was positive but of different magnitude according to the parity. When THI was below 62, an unfavourable effect was observed, especially in primiparous buffalo cows. A significant interaction between SCS and THI across parities was also observed. The effect of THI on MY across parities was not definite but overall a favourable relationship was observed. Our findings depict a susceptibility of buffaloes to low values of THI, suggesting an optimal THI range for water buffaloes between 59 and 63, although some deleterious effects were observed in primiparous buffaloes at THI values lower than 62. Additional investigations are needed to better elucidate the influence of THI on buffalo species.HIGHLIGHTS The overall effect of THI on buffalo diverges from what commonly observed in dairy cattle Cold stress affects milk and udder health in buffaloes The effect of THI on buffaloes’ performance depends on parity, with a larger susceptibility in primiparous than pluriparous buffalo cows Udder health in buffaloes, evaluated using somatic cell count, is also affected by TH

Autophagy is involved in granulosa cell death and follicular atresia in ewe ovaries

In mammalian ovaries, most follicles do not ovulate and are eliminated by atresia, which primarily depends on granulosa cell (GC) apoptosis. Autophagy is an alternative mechanism involved in follicle depletion in mammals through independent or tandem action with apoptosis. However, follicular autophagy has not yet been investigated in sheep; therefore, the present study aimed to investigate the involvement of autophagy in atresia among a pool of growing antral follicles in ewe ovaries. The abundance of the autophagic marker LC3B-II was determined using western blotting in GCs collected from ewe antral follicles. The antral follicles were classified as healthy or atretic based on morphological criteria and steroid measurements in follicular fluid (FF). Immunofluorescence and confocal microscopy analyses were performed on GCs to evaluate the presence of autophagic proteins and their subcellular localisation. Caspase-3 and DNA fragmentation were assessed using western blotting and TUNEL assays, respectively, in the same GC population to investigate the simultaneous apoptosis. The novel results of this study demonstrated enhanced LC3B-II protein expression in GCs of atretic follicles compared to that of healthy ones (1.3-fold increase; P = 0.0001, ANOVA), indicating a correlation between autophagy enhancement in GCs and antral follicular atresia. Autophagy, either functioning independently or in tandem with apoptosis, may be involved in the atresia of growing antral follicles in ewe ovaries because atretic GCs also showed high levels of apoptotic markers. The findings of this study might have important implication on scientific understanding of ovarian follicle dynamics

Validation of a radioimmunoassay method for cortisol in buffalo milk whey. A preparatory step for future sensor technology

One animal-based method to evaluate welfare is the presence of stress. In dairy ruminants, the responses to stressors include the activation of both neuroendocrine and autonomous nervous system that can be evaluated through an endocrine assessment. The present study aimed to validate a radioimmunoassay method for cortisol in buffalo milk. Three formulations (whole and skimmed milk and whey) and three solvents (methanol diethyl ether and dichloromethane) were tested: methanol was characterised by the best extraction efficiency (69.88%), whey cortisol concentrations showed a significant correlation with whole extracted milk and were not affected by fat content variation during the milking session. The RIA used in the present study showed good precision, sensitivity and specificity: the dilutions test indicated the high reproducibility of the results, overlapping of the dilution curve and standard curve highlighted high specificity and the lack of interfering factors by buffalo whey matrix. It is concluded that the present assay suits the cortisol measurement in buffalo milk and the ranges described could be employed in the calibration of a biosensing technologies directly integrated in milking parlour systems.Highlights Buffalo milk whey revealed to be a matrix of great interest because of its high stability in terms of storage, transportation and processing. RIA method suits the cortisol measurement in buffalo milk Ranges described can be employed in the calibration of biosensors for non-invasive assessment of cortiso

Incorporation of Testicular Ultrasonography and Hair Steroid Concentrations in Bull Breeding Soundness Evaluation

Testicular ultrasonography and steroid concentrations (cortisol, dehydroepiandrosterone sulfate (DHEA-S), cortisol/DHEA-S ratio, testosterone) in hair were examined for their utility in the bull breeding soundness evaluation (BBSE). Beef and dairy bulls (n = 16; 2.7 ± 0.4 years old; body condition score 3.2 ± 0.1) of five breeds were maintained under the same conditions at an accredited semen collection center. Bulls underwent routine semen collection twice weekly for 12 weeks and semen was processed and cryopreserved. Ultrasonography and hair sampling were undertaken at the last semen collection. Bulls with homogeneous testicular parenchyma (n = 8) had a higher (p n = 8). There were no differences (p > 0.05) in the hair concentrations of cortisol, DHEA-S, and testosterone between bulls with homogeneous and heterogeneous parenchyma. In bulls with homogeneous parenchyma, hair DHEA-S concentration was positively correlated with percentage motile sperm (R2 = 0.76), progressively motile sperm (R2 = 0.70), and motility yield (R2 = 0.71). The findings indicate that the integration of testicular ultrasonography and hair DHEA-S status in the BBSE could provide a more comprehensive assessment of indicative fertility in bulls. Additionally, ultrasonography can be used in the BBSE when the evaluation of semen parameters is not available