Giuseppe Piccione

Production of canine mesenchymal stem cells from adipose tissue and their application in dogs with chronic osteoarthritis of the humeroradial joints.

Autologous AD‐MSC [adipose‐derived MSC (mesenchymal stem cell)] therapy involves harvesting fat from the patient by isolating the stem and regenerative cells and administering the cells back to the patient. This study evaluated the production of canine AD‐MSCs and their possible application in cellular therapy for dogs. To assess whether cellular therapy can replace drug therapy, the clinical effect of a single intra‐articular injection of AD‐MSCs was evaluated on 4 dogs with lameness associated with OA (osteoarthritis) of the humeroradial joints. MSCs were readily isolated from adult dog adipose tissue, and their ability to form colony and differentiate into various phenotypes was confirmed. AD‐MSCs expressed OCT4, NANOG and SOX2 at the mRNA level, pluripotency markers usually ascribed to embryonic stem cells. The results suggest the stemness of the cells isolated from canine fat, and good quality control made them available for both experimental and clinical use. Follow‐up studies to evaluate the effects of AD‐MSC therapy showed that OA of the elbow joints improved with time, indicating significant potential for clinical use in the treatment of lameness, particularly when administered before the injury becomes severe.

The Circadian Rhythm of Body Temperature of the Horse

Rectal temperature of 10 female adult horses was recorded every 2 h for 10 consecutive days under a natural winter photoperiod (9 h of light and 15 h of darkness per day). A robust daily rhythm of body temperature was observed in all animals. The rhythm had a mean level of 38.3°C and a range of excursion of 1.0°C. Temperature started its daily ascent at dawn each day and reached a maximum 14 hours later. Body temperature of 5 of the horses was studied for 10 more days under constant illumination. The rhythm persisted under this condition, although with a slightly longer period of 24.2 h, which confirms the endogenous nature of the rhythm. Despite the fact that the body size of the horse is several orders of magnitude greater than that of rodents, the various parameters of the body temperature rhythm of the horse are similar to those of several species of rodents previously studied.

Daily and estrous rhythmicity of body temperature in domestic cattle

BackgroundRhythmicity in core body temperature has been extensively studied in humans and laboratory animals but much less in farm animals. Extending the study of rhythmicity of body temperature to farm animals is important not only from a comparative perspective but also from an economic perspective, as greater knowledge of this process can lead to improvements in livestock production practices. In this study in cattle, we investigated the maturation of the daily rhythm of body temperature in newborn calves, characterized the parameters of the daily rhythm in young cows, and studied the oscillation in body temperature associated with the estrous cycle in adult cows.ResultsWe found that the daily rhythm of body temperature is absent at birth but matures fully during the first two months of life. The mature rhythm had a mean level of 38.3°C, a range of excursion of 1.4°C, and was more robust than that of any mammalian species previously studied (90% of maximal robustness). Sexually mature cows also exhibited a robust estrous rhythm of body temperature. An elevation of about 1.3°C was observed every 21 days on the day of estrus. Small seasonal variations in this pattern were observed.ConclusionIn conclusion, calves exhibit a very robust daily rhythm of body temperature, although this rhythm is absent at birth and develops during the first two months of life. Adult cows exhibit also 21-day rhythmicity in body temperature reflecting the duration of the estrous cycle.

Analysis of serum proteins in clinically healthy goats (Capra hircus) using agarose gel electrophoresis

BACKGROUND Electrophoretic patterns of serum proteins provide useful information on pathological conditions in ruminants. Their reference values, however, are dissimilar to those of other species. Reference values for goats using agarose gel as the supporting matrix have not been reported. OBJECTIVE The aim of this study was to evaluate serum concentrations of total protein and protein fractions (albumin and globulins) by means of agarose gel electrophoresis (AGE) in goats in order to establish electrophoretic reference intervals and to evaluate potential changes associated with aging. METHODS Blood was collected from 105 clinically healthy Girgentana goats by means of jugular venipuncture. Serum protein concentrations were assessed by AGE. Three age groups were compared: 1-1.5 years, 2-4 years, and 5-12 years. RESULTS Values (mean ± SD) were determined for concentrations of total protein (72.26 ± 6.40 g/L), albumin (31.80 ± 4.00 g/L), α-globulins (6.40 ± 1.23 g/L), β(1) -globulins (10.50 ± 2.58 g/L), β(2) -globulins (5.18 ± 1.60 g/L), and γ-globulins (18.65 ± 5.90 g/L) and for albumin/globulin (A/G) ratio (0.82 ± 0.20). One-way ANOVA showed statistically significant age-related differences for total protein and α-globulin concentrations and A/G ratios. Age influenced protein concentrations with the 5-12-year-old group having higher total protein and α-globulin concentrations and lower albumin concentration and A/G ratios than the 2-4-year-old group. CONCLUSIONS This study provides reference values for total protein concentrations and protein fractions obtained by AGE in goats. Some values vary with age. Age-specific reference intervals are reported in order to provide clinicians with an additional diagnostic aid.

Reference intervals for total protein concentration, serum protein fractions, and albumin/globulin ratios in clinically healthy dairy cows.

The aim of the current study was to evaluate total serum protein concentration measured by the biuret reaction as well as albumin and globulin protein fractions determined by agarose gel electrophoresis. These data were used to establish reference intervals in dairy cows of different ages. Blood was collected from 111 clinically healthy Modicana dairy cows by means of jugular venipuncture. Reference intervals (mean ± standard deviation) were determined for total protein (67.54 ± 11.53 g/l), albumin (31.86 ± 4.60 g/l), α1-globulin (5.77 ± 2.20 g/l), α2-globulin (5.84 ± 1.90 g/l), β-globulin (7.46 ± 1.94 g/l), and γ-globulin (16.73 ± 4.54 g/l) concentrations as well as for albumin/globulin (A/G) ratio (0.88 ± 0.43). Values from 2-, 3-, 4-, 5-, and 6-year-old cows were compared statistically. One-way analysis of variance showed age-related differences for α-globulin and β-globulin fractions only. The results of the current study provide reference intervals for total protein concentration as well as albumin and globulin protein fractions in 2- to 6-year-old dairy cows.

Influence of Fasting and Exercise on the Daily Rhythm of Serum Leptin in the Horse

The hormone leptin is secreted by white adipocytes and regulates food intake and energy expenditure in rodents and humans. The goal of the present study was to investigate the existence of a daily rhythm of serum leptin in horses and its dependence on fasting and physical exercise. A robust daily rhythm of leptin was found in both athletic and sedentary horses, with a daytime trough and a peak in the dark phase. While physical exercise never induced changes in circulating leptin, fasting reliably affected serum leptin levels. Food deprivation did not abolish the daily rhythm of serum leptin, but daily mean leptin levels in fasted horses were significantly lower than in regularly fed horses. This result indicates that leptin production is not a mere consequence of feeding behavior. The fact that in a large animal such as the horse a short fast decreases leptin without significantly changing the body weight demonstrates that changes in levels of circulating leptin associated with food restriction do not solely reflect changes in amount of body fat.

Circadian modulation of starvation-induced hypothermia in sheep and goats.

Prolonged food deprivation is known to cause a fall in the core body temperature of homeotherms. In various species of small birds and mammals (body mass up to 2–3 kg), it has been shown that starvation-induced hypothermia is modulated by the circadian system, in the sense that hypothermia is observed primarily during the inactive phase of the daily activity cycle (i.e., during the night for diurnal animals and during the day for nocturnal animals), whereas relatively normal temperatures are recorded during the active phase. To investigate whether this modulation occurs also in larger animals, we investigated the effects of 4d food deprivation on the body temperature rhythm of goats and sheep (body mass 30–40 kg). In goats, the body temperature rhythm was found to have a mean level of 39.0°C with a mean daily range of excursion of 0.42°C. The daily oscillation in body temperature persisted during the first day of fasting, but the rhythm was drastically damped, if not eliminated, over the next 3 d as body temperature descended from the baseline level of 39.0 to 38.2°C. In sheep, the rhythm was found to have a mean level of 39.3°C with a mean daily range of excursion of 0.34°C. The daily oscillation in body temperature persisted through the 4 d of food deprivation, even though the mean level of body temperature gradually fell. Temperature fell more during the third and fourth nights than during the third and fourth days. Thus, circadian modulation of starvation-induced hypothermia was observed in sheep but not in goats.

Daytime profile of the intraocular pressure and tear production in normal dog

OBJECTIVE To evaluate the circadian rhythms of intraocular pressure (IOP) and tear production in dog exposed to a natural photoperiod. ANIMALS STUDIED We used 12 clinically healthy Beagles dog housed under natural photoperiod at indoor temperature and humidity. PROCEDURE Intraocular pressure and Schirmer tear test (STT) I were measured every 4 h over a 48-h period in both eyes in each animal. Statistical analysis of the data was performed by one-way repeated-measures anova, Students t-test, and single cosinor method. RESULTS On each day, there was a highly significant effect of time on both parameters. A statistically significant difference of STT I values was observed comparing left and right eyes (P < 0.0001). Robust daily rhythms were observed for both parameters, IOP values showed diurnal acrophase (left eye: 09:33 +/- 00:50 h; right eye: 09:25 +/- 00:22 h), while STT I values showed nocturnal acrophase (left eye: 20:27 +/- 00:46 h; right eye: 20:00 +/- 00:05 h). CONCLUSION This study has demonstrated circadian rhythms in both IOP and STT I.

Bioaccumulation of Heavy Metals in Blood and Tissue of Striped Mullet in Two Italian Lakes

Water and sediment from Faro and Ganzirri lakes were tested for cadmium, mercury, and lead. These heavy metals have been assessed in serum and muscles of Striped Mullet Mugil cephalus. A haemogram was performed to find effects of heavy metals on haematological variables. Students t-test showed higher Pb levels in sediments of Ganzirri Lake than Faro Lake. Two-way analysis of variance showed higher Cd and Pb concentration in muscles samples of Striped Mullet from Ganzirri Lake than from Faro Lake. Significant differences were found among haematological parameters as well. Mullet from Ganzirri Lake had lower white and red blood cell and trombocyte counts and higher mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration than those from Faro Lake. Our results indicate a significant presence of lead in the environmental produces an accumulation of this metal in fish muscles.

Daily rhythm of tear production in normal horse

OBJECTIVE This study was designed to assess the daily rhythm of tear production in clinically healthy horses, to compare left eye and right eye, and to evaluate the influence of time and gender. ANIMAL STUDIED For our study 18 horses (nine female and nine male) were subjected to three different light/dark (L/D) schedules: 12/12 L/D cycles, constant light (24/0 L/D) and constant darkness (0/24 L/D). PROCEDURES In all subjects Schirmer tear tests were performed at 4-h intervals over a 24-h period. RESULTS A circadian rhythm of tear production was found during the 12/12 L/D period and during constant darkness. Also our results showed a statistical significant difference comparing left eye and right eye, and between gender. CONCLUSIONS Our findings would suggest that the time of tear evaluation is significant in the diagnosis of tear film disorders, and that the difference between the two eyes and the two genders should be taken into consideration during the ophthalmic exam.