Mycoplasma mycoides subsp. mycoides SC identification by PCR in sperm of seminal vesiculitis-affected bulls.

In this study, by using the polymerase chain reaction (PCR) diagnosis for the detection and identification of Mycoplasma, we investigated mycoplasmas contaminating the semen of yearling bulls affected by seminal vesiculitis. The bulls presented neither subclinical nor clinical contagious bovine pleuropneumonia signs and the complement fixation test for specific antibodies was negative. Furthermore, we have investigated mycoplasmas isolated from semen of healthy breeding bulls of several breeds and origins, which routinely underwent breeding soundness examinations and presented no clinical signs of seminal vesiculitis. We were able to demonstrate mycoplasma infection in all tested samples by i) growth on mycoplasma-specific media and ii) a PCR-based method using a mycoplasma-specific MGSO/GPO1 primer set to amplify the 16S fragment rDNA. In addition, the identification of Mycoplasma species was made by PCR using the MSC1/MSC2 primer set that specifically amplifies M. mycoides subsp. mycoides SC or the MM450/MM451 primer set followed by AsnI digestion analysis in order to identify M. mycoides subsp. mycoides LC. The data presented herein clearly show that M. mycoides subsp. mycoides SC infection was associated with seminal vesiculitis while M. mycoides subsp. mycoides LC was only found in bull semen from healthy control animals. Our findings confirm that the M. mycoides subsp. mycoides SC is shed in the sperm making the ejaculate a valuable biological sample for the isolation of these bacteria from serologically negative animals. Although the pathogenic role of M. bovigenitalium in bull seminal vesiculitis has been established, our clinical findings, semen characteristics, microbiological and bacterial genomic analysis strongly suggest that M. mycoides subsp. mycoides SC may contribute to induce vesicular adenitis in the bull.     

Mammary cell turnover in lactating ewes is modulated by changes of energy fuels

The study of cell turnover of mammary gland helps to gain understanding of the subtle modulation that largely determines milk production. To evaluate the effect of diet composition on cell turnover of mammary gland, pluriparous sheep were allotted to three experimental groups and fed from day 33 after lambing a control diet (BD), a high starch (HS) diet or a high fat (HF) diet. Biopsies of mammary gland tissue were collected at 50 days after lambing for in situ detection of cell death and RT-PCR analysis of bax, bcl-2, caspase-3 and GST expressions. Both apoptotic and proliferation indexes were significantly higher (P<0.05) in the BD group compared to HS and HF groups. The relative expressions of GST were significantly greater (P<0.05) in HF group compared to the BD and HS groups. There was a significant increase in the ratio of bcl-2 to bax mRNA in the HS and HF groups. Availability of energy substrates for mammary gland can interfere with the cell fate, modulating genes involved in the control of oxidative stress which, in turn, can indirectly regulate cell apoptosis and proliferation.     

Apoptosis and Expression of Related Proteins in Mammary Gland of Heifers during Early Lactation

Veterinary Research Communications -     

Characterization of the complex carbohydrates in the zona pellucida of mammalian oocytes using lectin histochemistry

The objective of this study was to characterize the glycoconjugates present in the zona pellucida of the follicular oocytes in sheep, goats and pigs. The zona pellucida was stained with periodic acid-Schiff, low iron diamine, high iron diamine, and nine different lectin horseradish conjugates: Con-A, SBA, DBA, PNA, RCA-I, GSA-II, WGA, LTA and UEA-I. Staining with DBA, PNA, SBA and RCA-I was performed with and without saponification with KOH and sialidase digestion. The results showed the presence of neutral and acidic glycoconjugates with different terminal sugars and also sialic acid radicals in the zona pellucida of all the animals studied. In particular, the positive staining with WGA, SBA, PNA and RCA-I suggests the presence of oligosaccharides with N-acetyl-d-glucosamine and sialic acid linked to the penultimate -N-acetyl-d-galactosamine and to the disaccharide galactosyl-(1-3)-N-acetyl-d-galactosamine. The terminal trisaccharide sialic acid galactosyl-(1-4)-N-acetyl-d-glucosamine was identified only in the zona pellucida of ovine and procine oocytes. Thus, the zona pellucida exhibited species-specific variations in the content and distribution of lectin-binding patterns that may reflect the species specificity of gamete interaction.Abbreviations HID high iron diamine - HRP horseradish peroxidase - LID low iron diamine - PAS periodic acid-Schiff - PBS phosphate-buffered saline; see also Tables I and II     

Improving calving management to further enhance reproductive performance in dairy cattle

The aim of this study was to develop a system for the monitoring of calving to both reduce perinatal morta1ity and improve dairy cow fertility by preventing the majority of post-partum reproductive pathologies. Eighty dairy cows were assigned to the protocol of calving monitoring using GSM (Global System for Mobile Communications) technology. The application of GSM technology and the proper management of calving facilities comprise reliable approaches for calving assistance and improvements in reproductive efficiency and neonata1 viability. Based on the results of this study, we advocate the use of GSM technology on large farms for intensive mi1k production.     

Remote monitoring system as a tool for calving management in Mediterranean Buffalo heifers (Bubalus bubalis)

Buffalo breeding is common in Southern Italy. Dystocia compromises dam's and newborn health and welfare. Difficult parturition could be solved through prompt calving assistance, even if the identification of the beginning of delivery is challenging. Herein, we aimed to evaluate a remote calving alarm system in 15 Mediterranean buffalo heifers. An intravaginal probe was placed close to the external cervical os once premonitory signs of delivery were observed. No vaginal discharge nor signs of discomfort were notified in the days following the insertion of the probe. Heifers calved from 48 to 72 hr after the alarm was activated. The system correctly warned the farm personnel at the beginning of stage II of parturition, except for 2 cases. In the former, the intravaginal probe was expelled but the poor carrier network coverage negatively affected phone's signal quality; in the latter, recurrent vaginal prolapse was responsible for non-retention of the probe. Overall median expulsive phase was 68 ± 8 min, while the expulsion of a female calf took 54 ± 22.0 min and 90 ± 34.0 min in males, with significant difference (p =.02). Deliveries were homogeneously distributed across a 24-hr interval. No retention of foetal membranes nor metritis was identified at postpartum clinical examination. The calving alarm system used in this work was well tolerated in buffalo heifers. The introduction of smart technology in buffalo farming could contribute to the overall farm net return by reducing calf losses, especially for calves born from sexed-sorted semen, and by increasing animal welfare through quick resolution of dystocia. Further studies will be necessary to evaluate the net return in buffalo farms which will implement a remote calving alarm system on a wider population.     

Localization of the lectin reactive sites in adult and prepubertal horse testes

The testes of prepubertal and adult horses were investigated using 10 horseradish peroxidase conjugated lectins combined with sialidase digestion and potassium hydroxide treatment, to localise the oligosaccharide sequences of glycoconjugates during spermatid maturation. In adult animals, the lectins showed a variable affinity for spermatids and Sertoli cell apical extensions. Soybean agglutinin (SBA), peanut agglutinin (PNA), Ricinus communis agglutinin (RCA-I) and wheat germ agglutinin (WGA) bound to the acrosomal structures of spermatids, whereas Griffonia simplicifolia agglutinin (GSA-II) labelled these structures only during Golgi and cap phases. These results suggested that glycoproteins of mature acrosomes contain both N- and O-linked oligosaccharides and that these carbohydrate chains undergo modifications during spermiogenesis. Sialic acid residues were not detected throughout the acrosomal development. The lectin binding pattern of Sertoli cells was very similar to that of acrosome of spermatids during the maturation phase. In sexually immature horses, only the degenerated germinal cells and the Leydig cells showed reactivity towards lectins. The first cells reacted with SBA and Dolichos biflorus agglutinin (DBA), the latter with SBA, PNA, WGA, GSA-II, Canavalia ensiformis agglutinin (ConA), Lens culinaris agglutinin (LCA) and also with DBA after sialidase digestion.