Luigi Esposito

Effects of using vasectomized bulls in artificial insemination practice on the reproductive efficiency of Italian buffalo cows

The effects of the presence or absence of vasectomized male buffaloes on the reproductive efficiency of buffalo cows (n = 396) undergoing artificial insemination (AI) was studied on six farms owned and operated by a single consortium. Lactating animals were separated into two groups of various sizes on each farm and kept under semi-range conditions. Vasectomized bulls were present in one group at a bull/empty-cow ratio of 1:30. No bulls were present in the other group. Reproductive efficiency between the two groups over a period of 3.5 months was compared and evaluated on the basis of: 1) the number of spontaneous overt estruses associated with either feeble or intense signs of estrous behaviour; 2) the number of functional estrous cycles, i.e. estrous cycles with luteal phases defined as normal, based on specified progesterone concentrations in milk or blood plasma 8-10 days after estrus; 3) the number of consecutive functional estrous cycles in cases of induced estrus; and 4) pregnancy rate. Groups with bulls present demonstrated a significantly higher reproductive efficiency than groups without them. There was a higher incidence of spontaneous estrus (92 versus 69%; P < 0.01); spontaneous estrus of high intensity (62.2 versus 31.1%; P < 0.01); and higher incidence of functional estrous cycles following both spontaneous (65.8 versus 57.1%) and induced (77.0 versus 59.5%; P < 0.05) estrus. Exposure to vasectomised bulls also increased the incidence of consecutive functional estrous cycles (90.5 versus 68.1%; P < 0.01), and the pregnancy rate in cows inseminated at spontaneous (42.5 versus 18.9%; P < 0.01) or induced (51.1 versus 33.3%; P < 0.05) estrus. Overall pregnancy rate did not differ significantly between cows inseminated at induced or spontaneous estrus, although in the absence of bulls, pregnancy rate per AI was higher in cows inseminated at induced than at spontaneous estrus (33.3 versus 18.9%).

Effect of VDR polymorphisms on growth and bone mineral density in homozygous beta thalassaemia

Summary. We examined the effect of vitamin D receptor (VDR) polymorphisms at exon 2 (FokI) and intron 8 (BsmI) on the stature and bone mineral density at femoral neck (FBMD) and lumbar spine (LBMD) in 108 prepubertal and pubertal homozygous β thalassaemic patients, regularly treated. We found significantly shorter stature and lower LBMD and FBMD in all patients with CC VDR genotype, and significant shorter height and lower LBMD in prepubertal and pubertal female patients with BB VDR genotype. Because homozygous CC and BB VDR genotypes influence Vitamin D activity, they can be considered additional risk factors for bone disease in β thalassaemia.

Evaluation of leptin protein levels in patients with Cooley's anaemia

Endocrine disorders related to toxic effects of iron overload, such as short stature, delayed puberty and hypogonadism, represent major problems in both adolescent and adult thalassaemic patients. It has been suggested that gonadal dysfunction in these patients might be characterized mainly by a neuroendocrine dysfunction resulting in impaired hypothalaemic releasing hormone production, which is inadequate for proper pituitary stimulation (Kattamis & Kattamis, 1995). Pulsatile gonadotrophin releasing hormone (GnRH) treatment seems to re-establish, at least partially, the correct pituitary–gonadal function. Leptin, a recently discovered adipocyte-derived hormone, acts primarily at the hypothalamus, where it has effects on appetite, energy expenditure and the neuroendocrine axes. Leptin-deficient mice exhibit a complex phenotype which includes hypogonadism, impaired sexual maturation and infertility due to hypothalamic–pituitary hormone failure (Flier & Maratos-Flier, 1998). Exogenous leptin allows the recovery of the reproductive function. Recently, a related man and woman with congenital leptin deficiency have been described. Both showed hypogonadism (i.e. small penis and testis, primary amenorrhoea, respectively) of hypothalamic origin, suggesting that leptin not only controls body mass but may also be necessary to initiate human puberty (Strobel et al, 1998). To evaluate leptin levels in thalassaemic patients, blood samples were collected after an overnight fast from 162 patients with Cooley’s anaemia, 78 males (age: mean 19 6 7 years, range 5–39) and 84 females (age: mean 20 6 12 years, range 6–41) and from 138 normal subjects, matched for gender, age and body mass index (BMI). Leptin was measured by a commercial radioimmunoassay (Linco Research Inc., St Louis, Mo.). BMI was calculated as weight (kg) divided by height (m) squared. Mean leptin concentration was 2·69 6 1·23 ng/ml in thalassaemic males; this value appeared to be significantly lower (P <0·0005) than in controls (6·86 6 2·71 ng/ml). Although thalassaemic females also had lower leptin levels (6·37 6 2·9 ng/ml) than matched normal subjects (9·37 6 5·2 ng/ml), the differences were not significant (P<0·05). Furthermore, the physiologically positive BMI/ leptinaemia relationship, consistently found in normal and obese subjects, disappeared in thalassaemic males and was present, but lacked statistical significance in females (Fig 1). Therefore the adipocytes of thalassaemic patients seem unable to maintain adequate leptin production. The lack of any association between circulating leptin and body fat has already been observed in elderly subjects (Moller et al, 1998). Iron deposition studies in animal models have demonstrated that, although absent in the dermis, iron is well represented in subcutaneous layer adipocytes and that iron ions can seriously inhibit adipocyte activity (Youson & Sargent, 1984; Rejholcova et al, 1988). The differences between genders cannot be explained by an unequal iron overload (males, ferritin mean: 2·200 6 0·945 mg/l; females, ferritin mean: 1·938 6 0·848 mg/l; P <0·1), but could be the consequence of the physiologically different extension of the target organ (i.e. more subcutaneous adipose tissue in women). These results suggest that adipose tissue dysfunction can be considered as one of the endocrinopathies affecting thalassaemic patients. The consequent low leptin levels might play a role in the neuroendocrine dysfunctions reported in these patients.

HEMATOLOGICAL AND MOLECULAR ANALYSIS OF β-THALASSEMIA AND Hb LEPORE IN CAMPANIA, ITALY

This epidemiological study was based on a hematological and a molecular analysis of 310 heterozygous β thalassemic and 75 carriers of Hb Lepore out of 3,000 microcythemic subjects from the Campania region of Italy. The molecular analysis of β chains and the δβ hybrid gene has shown different β chain defects, but only the Hb Lepore-Boston-Washington type in association with haplotypes I and V. The prevalence and distribution of these molecular defects in Campania show that they are linked to historical events and to the geographical characteristics of this region.

Influence of oocyte donor on in vitro embryo production in buffalo

The aim of this research was to estimate the variability between buffalo as oocyte donors. In Experiment 1, reproductive variables were retrospectively analyzed in buffalo (n=40) that underwent repeated ovum pick up (OPU), over 16 puncture sessions (PS). The follicular recruitment among individuals and the relationship between follicular population and oocyte production were evaluated. In Experiment 2, eight buffalo underwent OPU for 28 PS and the oocytes were processed separately to correlate follicular and oocyte population at the first PS to blastocyst (BL) production. In Experiment 1, the average number of total follicles (TFL), small follicles (SFL), cumulus-oocyte complexes (COC) and Grade A+B COC recorded in each 4-PS period had great repeatability (r=0.52, 0.54, 0.60 and 0.57, respectively). The average number of Grade A+B COC recovered during the subsequent 15 PS was positively correlated with the first PS number of TFL (r=0.60; P<0.001), SFL (r=0.68; P<0.001), COC (r=0.48; P<0.01) and Grade A+B COC (r=0.40; P<0.05). In Experiment 2, a large variability among animals was observed in blastocyst yields. When animals were grouped according to the BL yield, the greatest BL yield group had a greater (P<0.05) number of TFL (8.3 ± 0.9 compared with 5.6 ± 0.7) and SFL (7.3 ± 0.3 compared with 3.8 ± 0.7) at the first PS than the lesser BL yield group. The average number of BL produced over the subsequent sessions was correlated with the number of TFL (r=0.80; P<0.05) and COC (r=0.76; P<0.05) observed at the first PS. These results demonstrated a donor influence on the oocyte and BL production, suggesting a preliminary screening to select the donors with greater potential.

A CellBE-based HPC Application for the Analysis of Vulnerabilities in Cryptographic Hash Functions

After some recent breaks presented in the technical literature, it has become of paramount importance to gain a deeper understanding of the robustness and weaknesses of cryptographic hash functions. In particular, in the light of the recent attacks to the MD5 hash function, SHA-1 remains currently the only function that can be used in practice, since it is the only alternative to MD5 in many security standards. This work presents a study of vulnerabilities in the SHA family, namely the SHA-0 and SHA-1 hash functions, based on a high-performance computing application run on the MariCel cluster available at the Barcelona Supercomputing Center. The effectiveness of the different optimizations and search strategies that have been used is validated by a comprehensive set of quantitative evaluations, presented in the paper. Most importantly, at the conclusion of our study, we were able to identify an actual collision

Acepromazine-dexmedetomidine-ketamine for injectable anaesthesia in captive European brown hares (Lepus europaeus)

OBJECTIVE To evaluate a combination of acepromazine, dexmedetomidine and ketamine (ADK) on induction and recovery from anaesthesia, and on physiological parameters in hares undergoing non-invasive procedures. STUDY DESIGN Prospective clinical study. ANIMALS Sixteen European hares (Lepus europaeus), seven males and nine females, aged (mean ± SD) 3.25 ± 0.9 months and weight 2.1 ± 0.6 kg. METHODS Acepromazine 1% (A), dexmedetomidine 0.05% (D) and ketamine 5% (K) were mixed and given intramuscularly (IM) at 0.25 mL kg(-1), representing 10 mg kg(-1) K, 0.25 mg kg(-1) A, 12.5 μg kg(-1) D. If the righting reflex was present after four minutes, a second injection of 0.15 mL kg(-1) (6 mg kg(-1) K, 0.15 mg kg(-1) A, 7.5 μg kg(-1) D) was administered IM. Surgical anaesthesia was judged as present when righting, palpebral, ear-pinch and pedal withdrawal reflexes were absent. Anaesthetized hares were tagged, and underwent blood sampling and ocular ultrasound examination. Physiological parameters were recorded every ten minutes, and were compared by Kruskal-Wallis tests. RESULTS A single dose induced loss of righting reflex in 11/16 (69%) hares within four minutes; the second dose was effective in the remaining hares. Ten minutes after the loss of the righting reflex, a surgical plane of anaesthesia was present in all hares. Sleep time to regaining righting reflex was 34 ± 11 (range 21-62) minutes and recovery was calm. Although there were some statistical differences over time, cardiovascular parameters remained within an acceptable range but there was respiratory depression and hares were hypoxemic. CONCLUSIONS AND CLINICAL RELEVANCE The ADK mixture produced a smooth and rapid induction of anaesthesia, a low incidence of untoward side effects and full recovery after four hours. Supplementary oxygen might be advisable if a deeper plane of anaesthesia was required. Chemical restraint was adequate to perform non-invasive procedures.

Reactive Oxygen Species (ROS) and Male Fertility

Oxidative energy production is inevitably associated with the generation of reactive oxygen species (ROS), excessive concentrations of which can lead to cellular pathol‐ ogy. A free radical may be defined as any molecule that has one or more unpaired electrons. The superoxide anion, the hydroxyl radical, and the hypochlorite radical are some of the highest reactive radicals of oxygen. Owing to their high reactivity and to their capability of initiating an uncontrolled cascade of chain reactions, ROS produce extensive protein damage and cytoskeletal modifications and inhibit cellular mechanisms. Aerobic organisms are equipped with a powerful battery of mechanisms that protect them from the adverse effects of lipid peroxidation (LPO) and other manifestations of oxygen toxicity. Defective sperm function frequently causes male infertility, due to abnormal flagella movement, failure to recognize the zona, and inhibition of sperm-oocyte fusion. ROS are fundamental mediators of physiological sperm function, such as signal transduction mechanisms that have an effect on fertility. ROS can have positive effects on sperm and the concentration functions depending on the nature and the concentration of the ROS involved. They are necessary in regulating the hyperactivation and the ability of the spermatozoa to undergo acrosome reaction. An increased amount of superoxide anion (O2) is one of the first steps required by the spermatozoa for induction and development of hyperactivation and capacitation. Numerous studies have shown that oxidative stress plays an important role in the pathophysiology of infertility and assisted fertility. The paternal genome is of primary importance in the normal embryo and fetal develop‐ ment. ROS-induced sperm damage during sperm translation, such as signal trans‐ duction through the seminiferous tubules and epididymis, is one of the most important mechanisms leading to sperm DNA damage. Male germ cells are extremely

Effect of pour-on alphacypermethrin on feed intake, body condition score, milk yield, pregnancy rates, and calving-to-conception interval in buffaloes

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.

Helminths of Wild Boar (Sus scrofa) in the Calabrian Region of Southern Italy

Abstract: In Calabria, as in other Italian regions, the wild boar (Sus scrofa) population has increased considerably in the last few years. The presence of this wild ungulate, a huntable species, was detected throughout this region. Wild boars are a host for many endoparasites, some of which are zoonotic. Our aim was to acquire data on the prevalence endoparasites in wild boar in the region of Calabria, Italy, where such information is limited. We used 60 wild boars culled during the 2014–15 hunting season. We used FLOTAC dual techniques for fecal microscopic exams, as well as qualitative and quantitative microscopic examination with a sensitivity level of 2 eggs/g of feces. The research has underlined the remarkable diffusion of gastrointestinal helminths, particularly Ascaris suum, in wild boar of the areas of Sila and Presila in the province of Catanzaro, Calabria.