Chronophysiology of domestic animals.

This review highlights recent findings on biological rhythms and discusses their implications for the management and production of domestic animals. Biological rhythms provide temporal coordination between organs and tissues in order to anticipate environmental changes, orchestrating biochemical, physiological and behavioural processes as the right process may occur at the right time. This allows animals to adapt their internal physiological functions, such as sleep-wake cycles, body temperature, hormone secretion, food intake and regulation of physical performance to environmental stimuli that constantly change. The study and evaluation of biological rhythms of various physiological parameters allows the assessment of the welfare status of animals. Alteration of biological rhythms represents an imbalance of the state of homeostasis that can be found in different management conditions.

Using infrared thermography for the evaluation of road transport thermal homeostasis in athletic horse.

This study aimed to evaluate changes in body temperature in athletic horses during two different road transport distances. Six Italian Saddle horses were subjected to a 100 and 300 km transport during different times of day (am and pm). Rectal and cutaneous temperatures were recorded before (T0), immediately (following 5 min- T1) and 1 hour (T2) after transport by means of a rectal digital thermometer and a thermal infrared camera (FLIR T440) respectively, for the evaluation of left and right side of four body regions: jugular, shoulder, croup and inner thigh. There were no differences between left and right sides, inner thigh or rectal temperatures when comparing the transport distance, time points or time of day. At T0, jugular (P < 0.0001), shoulder (P < 0.01) and croup (P < 0.01) average temperatures were higher in the pm compared to those in the am in both journeys. At T1, jugular (P < 0.01) and croup (P < 0.01) temperatures were lower in the pm compared to am following the 300 km journey. Jugular temperature (P < 0.0001) was higher following the 300 km compared to the 100 km journey at each time point (T1 and T2) at both times of day (am and pm). Shoulder (P < 0.0001) and croup temperatures (P < 0.0001) were higher at T2 after the 300 km journey than at T2 after the 100 km journey). The current results suggested a difference between the two distances and the time of day appeared to have as great effect on ocular temperature as road transport distance.

Monitoring of inflammatory blood biomarkers in foals with Rhodococcus Equi pneumonia during antimicrobial treatment.

Rhodococcus equi (R. equi), a gram-positive facultative intracellular pathogen, is a common cause of pneumonia in foals and represents a major cause of disease and death. The aim of the present study was to investigate the time-depended changes in White Blood Cells (WBC), basophils (Baso), neutrophils (Neu), lymphocytes (Lymf), monocytes (Mon), eosinophils (Eos), platelet (PLT) counts, fibrinogen (Fbg) concentration, interferon (IFN-α, IFN-γ) and interleukins (IL-2 and IL-10) in foals with clinical R. equi pneumonia. The main treatment was with azithromycin-rifampicin for 14 days. Blood was sampled prior to, 7 and 14 days after starting therapy. Treatment was associated with significantly decreased counts of WBC, (25.6 ± 6.7 and 14.2 ± 2,7 × 103/ml), Neu (18.6 ±6.2 and 10.7 ± 3.1 × 103/ml), Mon (1.5 ± 0.5 and 0.9 ± 0.2 × 103/ml) and Fbg (539 ± 124 and 287 ± 26 g/dl) between day 0 and day 14. IL-2 and IL-10 concentrations were significantly increased (P = 0.028, P = 0.013, respectively) after treatment, whereas IFN-α and IFN-γ concentrations were not. The diagnostic potentials of INF-α, INF-γ, IL-2 and IL-10 per se seems not very high, however, the study suggests that the activity change of selected interleukins in the course of the disease may be associated with amelioration. We concluded that patterns of serum concentration changes of INF-α, INF-γ, IL-2 and IL-10 may help in the study of the innate immune response in foals during infection and treatment of R. equi pneumonia.

Potential Implications of Acid-Sensing Ion Channels ASIC2 and ASIC4 in Gonadal Differentiation of Dicentrarchus labrax Subjected to Water Temperature Increase during Gonadal Development.

In this study, the expression and implication of acid-sensing ion channels 2 and 4 (ASIC2 and ASIC4) in the gonadal sex differentiation of Dicentrarchus labrax (D. labrax), subjected to increasing water temperatures during gonadal development, were evaluated. Two groups were selected: a control group (CG), in which the average water temperature was maintained at 15 °C and increased to 20 °C in 20 days until weaning; and an experimental group (EG), in which the water temperature was retained at 15 °C for 60 days; thereafter, the temperature was increased daily by 0.5 °C until it reached 20 °C up to the weaning time. Ten fish from the CG and 13 fish from the EG were sampled randomly on the 335th day after hatching (dph). A higher percentage of gonad differentiation in ovaries rather than in testes was observed in the EG compared to the CG (p = 0.01). ASIC2 and ASIC4 were detected for the first time in D. labrax ovaries by indirect immunofluorescence. Both ASIC2 and ASIC4 were expressed in previtellogenic oocytes of ovaries and in scattered cells within some testes, and were most likely intratesticular previtellogenic oocytes in both the CG and EG groups. The CG group showed a higher expression of ASIC4 than the EG cohort (p < 0.05). The results gathered in this study revealed the capacity of water temperature to influence both gonadal differentiation and growth in this gonochoristic fish species, and suggests the possible role of ASIC2 and ASIC4 in gonad differentiation and gamete development in D. labrax.

Blood and hoof biodistibution of some trace element (Lithium, Copper, Zinc, Strontium and, Lead) in horse from two different areas of Sicily.

BACKGROUND AND PURPOSE: Biological monitoring of trace element horses is a well-known tool for investigating potential bioaccumulation in urbanized and industrialized geographical areas. Some biomaterials such as hoof are considered as an important indicator of environmental pollution. Hooves can store trace elements for a long time compared to blood and this represents a scientific key to long-term monitoring of exposure to environmental pollutants. In the present study, samples of equine hoof and blood were taken from an experimental group of horses living in an industrialized area of Sicily (Italy) and from a control group of horses housed in a non-industrialized area of Sicily to evaluate the bioaccumulation of different trace elements in different substrates. METHODS: Hoof and blood mineral concentration of Li- Lithium, Cu- Copper, Zn- Zinc, Sr- Strontium and, Pb- Lead was processed by means of Thermo Scientific iCAP-Q ICP-MS spectrometer. Student T-test was applied to evaluate the differences between groups and substrates. Correlation analysis between substrates and hematological parameters was performed. RESULTS: Statistical analysis showed a significant statistical difference for Li (p < 0.0001) and Pb (p < 0.0001) between blood and hoof in both groups. A statistically significant difference was observed for Li (p < 0.0001), Cu(p < 0.01), Zn (p < 0.0001) and Pb (p < 0.0001) between substrates in the experimental group. The statistical analysis showed a significant difference of blood Zn (p < 0.01) and hoof Pb (p < 0.01) concentration between groups. Our results revealed a statistically higher concentration of minerals in the blood substrate than in the hoof in both groups. Li (p < 0.0001) and Zn (p < 0.001) were positively correlated between control and experimental group in blood substrate. No correlations were found (P > .05) in hoof between groups for other elements. A positive correlation with WBC was observed in Cu concentration for hoof substrate (p < 0.02) in control group and for Zn in experimental group (p < 0.02). A positive correlation with HGB and HCT was observed for Cu hoof concentration (p < 0.02) in experimental group and PLT resulted positively correlated (p < 0.02) with Pb blood substrate in control group. CONCLUSIONS: Horses were not overexposed to these elements in the analyzed groups. The study highlights the role of a new bioaccumulation substrate such as the horse hoof identified as an analytical matrix for the monitoring of heavy metal concentrations in domestic animals.

Horse Whole Blood Trace Elements from Different Sicily Areas: Biomonitoring of Environmental Risk.

Horses are excellent bioindicators for the assessment of environmental pollution. The aim of this study was to evaluate the levels and potential bioaccumulation of 28 mineral elements in 75 horse whole blood samples collected from five pollution-prone areas of Sicily, Italy. A direct mercury analyzer (DMA-80) was used for Hg determination, and an inductively coupled plasma mass spectrometer (ICP-MS) for all other elements. A one-way ANOVA test, followed by Bonferroni's multiple comparison for post hoc comparison, was applied to assess statistically significant differences between mineral elements and the five experimental groups. The levels of mineral elements in hay and concentrate were below the limits set by Regulation No. 744/2012. The mineral content of whole blood samples was slightly influenced by the region of origin of the horse. p values < 0.05 were statistically meaningful. However, the concentrations of mineral elements in horses' whole blood remained within reference ranges. In conclusion, the present study shows that the mineral content does not represent a toxicological risk for the analyzed horses. In addition, the study areas did not appear to show a high mineral element contamination.

Circadian Variation of Peripheral Blood Cells in Horses Maintained in Different Environmental and Management Conditions.

The aim of our study was to analyze circadian rhythm of the hematological profile of horses housed in a loose box and paddock during the different seasons (spring, summer, autumn, and winter). Blood samples were performed every 4 h for 48 consecutive hours. Red blood cells (RBCs), hemoglobin (HGB), hematocrit (HCT), white blood cells (WBCs), platelets (PLTs), and leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes) were analyzed, and, at the same time, environmental conditions were recorded. A statistically significant effect of housing conditions (p < 0.0001) was observed on all hematological values except for WBC during winter and for neutrophils (p < 0.0001) during spring and autumn. A statistically significant effect of season (p < 0.0001) was found for RBC, HCT, and PLT and for all leukocyte cells (p < 0.0001) except for basophils. The single Cosinor method revealed a daily rhythm of hematological parameters during spring in both groups, and a daily rhythm for lymphocytes and neutrophils was observed during spring and summer in horses kept in a loose box and during winter in horses housed in a paddock. Our results revealed that the response of the immune system is regulated by circadian physiology. Knowledge of the periodic temporal structure of mammals should be considered when evaluating animals' adaptation to temporizations imposed by the environment.

MiR-148a-3p Promotes Colorectal Cancer Cell Ferroptosis by Targeting SLC7A11.

Ferroptosis, an iron-dependent form of cell death, and dysregulated microRNA (miRNA) expression correlate with colorectal cancer (CRC) development and progression. The tumor suppressor ability of miR-148a-3p has been reported for several cancers. Nevertheless, the role of miR-148a-3p in CRC remains largely undetermined. Here, we aim at investigating the molecular mechanisms and regulatory targets of miR-148a-3p in the CRC cell death mechanism(s). To this end, miR-148a-3p expression was evaluated in SW480 and SW620 cells and normal colon epithelial CCD 841 CoN cells with quantitative real-time polymerase chain reaction (qRT-PCR). Data reported a reduction of miR-148a-3p expression in SW480 and SW620 cells compared to non-tumor cells (p < 0.05). Overexpression of miR-148a selectively inhibited CRC cell viability (p < 0.001), while weakly affecting normal CCD 841 CoN cell survival (p < 0.05). At the cellular level, miR-148a-3p mimics promoted apoptotic cell death via caspase-3 activation (p < 0.001), accumulation of mitochondrial reactive oxygen species (ROS) (p < 0.001), and membrane depolarization (p < 0.001). Moreover, miR-148a-3p overexpression induced lipid peroxidation (p < 0.01), GPX4 downregulation (p < 0.01), and ferroptosis (p < 0.01), as revealed by intracellular and mitochondrial iron accumulation and ACSL4/TFRC/Ferritin modulation. In addition, levels of SLC7A11 mRNA and protein, the cellular targets of miR-148a-3p predicted by bioinformatic tools, were suppressed by miR-148a-3p's overexpression. On the contrary, the downregulation of miR-148a-3p boosted SLC7A11 gene expression and suppressed ferroptosis. Together, these in vitro findings reveal that miR-148a-3p can function as a tumor suppressor in CRC by targeting SLC7A11 and activating ferroptosis, opening new perspectives for the rationale of therapeutic strategies through targeting the miR-148a-3p/SLC7A11 pathway.

Comparison of the Faecal Microbiota Composition Following a Dairy By-Product Supplemented Diet in Nero Siciliano and Large White × Landrace Pig Breeds.

The current study compared the faecal microbiota composition of two pig breeds (autochthonous vs. commercial) to understand what happens after the integration of liquid whey in the diet and what the role of the host genetic is. The trial was conducted for 60 days, and the faecal microbiota composition was investigated at three time points, T0, T1 (after 30 days) and T2 (after 60 days) in 30 female pigs (20 commercial crossbred and 10 Nero Siciliano pigs). The animals were divided into four groups (two control and two treatment groups). Generally, in both breeds, Firmicutes (51%) and Bacteroidota (36%) were the most abundant phylum whereas Prevotella, Treponema and Lactobacillus were the most abundant genera. The two breeds have a different reaction to a liquid whey diet. In fact, as shown by PERMANOVA analysis, the liquid whey significantly (p < 0.001) affects the microbiota composition of crossbreeds while not having an effect on the microbiota of the Nero Siciliano. Despite this, in both breeds Bifidobacterium and Ruminococcus have been positively influenced by liquid whey and they promote intestinal health, improve immunity, increase performance, and feed efficiency. In conclusion, the integration of liquid whey had a different effect on the Nero Siciliano and crossbred pig breeds, emphasizing the importance of the host genetic profile in determining the faecal bacterial composition.

Cardiac Biomarker Responses to Acute Exercise in Show Jumping Horses.

Cardiac biomarkers are useful to identify cardiac muscle variations in human and equine medicine. The aim of this study was to investigate the acute effect of a bout of show jumping training on serum activity of cardiac and muscular biomarkers in healthy athletic horses to include cardiac troponin (cTnI), myoglobin (MB), aspartate amino transferase (AST), alanine amino transferase (ALT), creatine phosphokinase (CPK) and lactate dehydrogenase (LDH). Serum samples were collected from seven Italian Saddle horses (three geldings and four mares; 10 ± 3 years; mean body weight 480 ±70 kg), regularly trained for show jumping at rest, immediately after exercise (show jumping simulate trial) and during the recovery period (30 and 60 min after exercise). ANOVA was applied to all parameters, and Pearson correlation coefficient t (r) evaluated. Immediately after exercise there was an increase in cTnI (P < .01), MB (P < .01), and CPK (P < .005); a positive correlation between cTnI and AST and between AST and LDH; and a negative correlation between cTnI and ALT and between ALT and CPK. Thirty minutes after exercise, there was a positive correlation between AST and ALT and between AST and LDH, while 60 mintues after exercise, there was a positive correlation between MB and LDH and a negative correlation between AST and CPK. The results obtained demonstrate the cardiac and muscular response to short-term intense exercise show jumping exercise.

The role of embryonic stem cells, transcription and growth factors in mammals: A review.

Mammals represent a relevant species in worldwide cultures with significant commercial value. These animals are considered an attractive large animal model for biomedical and biotechnology research. The development of large animal experimental models may open alternative strategies for investigating stem cells (SCs) physiology and potential application in the veterinary field. The embryonic stem cells (ESCs) are known to possess natural pluripotency that confers the ability to differentiate into various tissues in vivo and in vitro. These notable characteristics can be useful for research and innovative applications, including biomedicine, agriculture and industry. Transcription factors play a crucial role in preserving stem cell self-renewal, whereas growth factors are involved in both growth and differentiation. However, to date, many questions concerning pluripotency, cellular differentiation regulator genes, and other molecules such as growth factors and their interactions in many mammalian species remain unresolved. The purpose of this review is to provide an overall review regarding the study of ESCs in mammals and briefly discuss the role of transcription and growth factors.

Simultaneous recording of subcutaneous temperature and total locomotor activity in Bos taurus and Bos indicus raised in a subtropical region of Argentina.

The aim of this study was to evaluate some physiological parameters as total locomotor activity and subcutaneous temperature in two different bovine breeds under tropical environment. In this study, total locomotor activity and subcutaneous temperature were evaluated in 18 clinically healthy female cows. All cows were divided into 3 groups: group A1 was constituted by 6 Holstein Bos taurus, group A2 was constituted by 6 female Herford Bos taurus cows, and group B was constituted by 6 female Bradford Bos indicus cows. Data recording of total locomotor activity (TLA) and subcutaneous temperature (ST) was performed by means of a subcutaneous thermometer (Thermocron) and an actimeter ACTIWATCH® (Cambridge Neurotechnology Ltd.) based on accelerometer technologies equipped on all subjects to record TLA. At the same time, thermal and hygrometric recording were carried out, considering the subtropical climate of Santa Fe. The application of GLM for statistical analysis showed a significant effect (P < 0.05) on statistical model and time of the day on TLA and ST for all groups; no significative effects on animal parameters were found for ST and TLA except for group B. Circadian parameters have been evaluated according to the single cosinor procedure of ST that showed a diurnal daily rhythmicity for all investigated groups and TLA which is focused almost during the photophase for groups A1 and A2 and during scotophase for group B. Considering different species and breeds, and different environmental conditions, this study suggested that some subjects may be much more able to adapt themselves to environmental stress than others. During their evolution from Bos taurus, zebu cattle (Bos indicus) have acquired genes that confer thermotolerance. Subjects from Bos indicus breeds are better able to regulate body temperature in response to heat stress than Bos taurus subjects.

Effects of the Ingestion of Ripe Mangoes on the Squamous Gastric Region in the Horse.

Erosions and gastric ulcers may be present in horses at any age and under different conditions of rearing and handling. In tropical regions, horses can feed on fruits rich in soluble carbohydrates, such as mangoes, but little is known about how these foods interact with their digestive systems. To test the hypothesis that the ingestion of ripe mangoes with peels could cause disturbances in the digestive processes of horses, an experiment was developed to monitor animals that had free access to ripe mangoes in their pasture areas. Horses (purebred Arabians, n = 5; ~340 kg, ~13 years) were evaluated by video gastroscopy and blood analysis. A controlled postprandial glucose curve for mango intake was also performed. Gastroscopies were performed at intervals of 15 days, starting in December, just before the beginning of the harvest, until the beginning of February, and days after the end of the harvest. Blood collection was performed on the same day between November and February for blood analysis. The results were submitted to ANOVA and Tukey's test, with a significance level of p < 0.05. Gastroscopies indicated that four out of five horses had erosions and ulcers in the squamous region between 15 and 30 days after the start of the season. Biochemical tests indicated a reduction in plasma proteins during the harvest period, and the postprandial glucose curve showed concentrations above 200 mg/dL between 30 and 180 min after ingestion of 5.37 kg mangoes. The animals were not treated and recovered after 15 days of harvest and without ripe mangoes on the ground. It is concluded that the indiscriminate ingestion of mangoes favors the appearance of lesions in the gastric squamous region, to varying degrees, and that animals recover naturally after an average of 15 days from the end of the season when the animals return to their regular feeding with hay and grass pasture.

Correlation between Ocular and Rectal Temperature with Intra Ocular Pressure in Horse during Exercise.

The aim of the present study was to investigate the response to physical exercise of some ocular physiological variables (ocular temperature and intraocular pressure) in horses performing a jumping course in the morning (a.m.) or in the afternoon hours (p.m.), before and after the exercise, in correlation with the rectal temperature. Data collection was carried out on 14 clinical healthy Italian saddle horses. All horses were trained from 9:00 to 10:00 a.m. and from 19:00 to 20:00 p.m. according to a specific training program. Ocular temperature (OT), rectal temperature (RT) and intraocular pressure (IOP) were determined. Statistical analysis showed no differences between the results for the left and the right eyes. The application of two-way repeated-measures analysis of variance (ANOVA) showed a statistically significant effect of time (before vs. after) on IOP (p = 0.0001). RT and OT were statistically influenced by time of day and by experimental conditions (a.m. vs. p.m.) (p = 0.0001). Bland-Altman (B-A) testing showed an agreement between the values of RT and OT. Our results showed a correlation between increased body temperatures and IOP variations during exercise, so they can probably be considered indicators of athletic horses' physical fitness during exercise.

Lithium Concentration in Biological Samples and Gender Difference in Athletic Horses.

Lithium (Li+) represent a substance particularly used for human psychiatric disorders but its therapeutic effect is not well documented for equine specie. For its neurotrophic effect, it may be used as a possible doping substance in horses sports competitions. The purpose of the present study was to determine the different bioaccumulation of lithium concentrations in different biological substrates (blood, serum, mane, and tail), in 30 horses (15 geldings and 15 mares) and hematological parameters as blood biomarkers for lithium bioaccumulation. (RBC, WBC, HGB, HCT, MCV, MCH, MCHC). The lithium concentration in substrates were analyzed by one-way ANOVA. An unpaired t-test was performed between geldings and mares in all substrates. Furthermore, a single regression analysis (Pearson) has been carried out between the lithium concentrations of all biological substrates with each other and between the substrates with the hematological parameters. Our results showed a higher value of lithium concentration in blood and mane in total, and a higher value in mane and tail for mares than geldings. Correlation analysis showed only a significant correlation among blood, serum, and tail. A significates statistical correlation between blood and all hematological parameters except for WBC and MCH was observed. A significant correlation was shown for serum and RBC, HGB and HCT, and between tail and HGB, HCT and MCHC. No significant correlation was observed for mane and hematological parameters. These data could be useful for the evaluation of biodistribution of lithium in an athletic horse in relation to different gender for future use in doping control.

Diurnal variation in rectal and cutaneous temperatures of horses housed under different management conditions.

Thermoregulation is an important mechanism to ensure thermal homeostasis. In horses, different housing conditions could influence this mechanism by exposing animals to different microclimates. The circadian rhythm of body temperature is well known. For this reason, we wanted to investigate the daily rhythm of cutaneous (jugular, shoulder, inner thigh and croup) temperature in comparison to rectal temperature in athletic horses kept under two different housing conditions. Ten clinically healthy Italian Saddle horses were divided into two equal groups: the first group was housed in individual boxes, and the second group was kept in paddocks. In all horses, rectal and cutaneous temperature measurements were performed every 4 h for 48 consecutive hours during different seasons (spring, summer, autumn and winter), respectively, by means of digital and infrared thermometers. Ambient temperature, relative humidity and ventilation were recorded simultaneously. A general linear model (GLM) was applied to the recorded temperature values displaying a significant effect of season, time of day, site of recording and housing condition (p < 0.001). No statistical differences were found between the two days of monitoring (p = 0.49). A trigonometric statistical model (single cosinor method) was applied to investigate the circadian rhythm of rectal and cutaneous temperatures in the two different groups. Our results showed circadian rhythmicity of rectal temperature during all seasons and in both groups. Cutaneous temperature shows daily rhythmicity that was different in the various regions and was influenced by housing conditions and seasons. Application of GLM also showed a statistically significant effect of season and site of recording (p < 0.0001) on all circadian parameters and of management condition on amplitude and robustness (p < 0.0001). No statistical differences between the two days of monitoring were observed (p = 0.68). These findings are probably due to the animals' thermoregulatory mechanisms ensuring heat exchange between the body and the environment, and that was influenced by the microclimate. In particular, the microclimate influenced the thermodispersion disrupting the daily rhythmicity of some body regions, in horses kept in boxes. Only the temperate climatic conditions observed in summer guarantee the expression of the daily rhythmicity of all body surfaces in both management conditions. These results contribute to the knowledge of the mechanisms of homeostasis and control of body temperature in the athletic horse kept in different housing conditions, with a chronophysiological interpretation that completes the proper management of thermal well-being in horses.

How Much Energy Vaquejada Horses Spend in a Field Simulation Test?

Vaquejada is a high-intensity and short-duration exercise in which helper horses (HH) are responsible to keep a bull running in a line while pull horses (PH) work to put the bull down after 100 m of running. The purpose of this study was to quantify and compare energy expenditures (EE), transport costs (COT) and metabolic energy requirements (Pmet) of horses used in Vaquejada. Thus, eight Quarter Horses, in randomly formed pairs, performed a vaquejada simulation test (VST), which consisted of three races on a sand track (130-150 m), with a 5-min interval between them. All horses used an integrated heart rate (HR) and GPS monitoring system (V800, Polar Electro) and, from these data, EE, COT and Pmet were calculated using the formulas: EE (J/kg/min) = 0.0566 × HR1.9955, COT = (HR-35)/kg/m × 103 and Pmet = (HR-35)/min/kg. Blood samples were collected for lactate analysis at rest, immediately after the first, second and third race and after 30 min of recovery. Data obtained were submitted to one-way ANOVA and Tukey tests (p ≤ 0.05). In VST, HH had higher EE and higher HR at trot; while PH presented higher EE and HR at canter. Finally, considering total VST, PH had higher EE and COT, while HH had higher Pmet. Lactate was higher in PH. Despite practicing the same sport, PH and HH should be considered distinct athletes, and these must be considered to set up appropriate physical and nutritional programs, which will lead to better performance and guarantees of well-being.