Protein and energy relationships in dairy cattle. 1. Dry cows

Nitrogen metabolism in ruminants is a dynamic process depending on level of intake and composition of dietary dry matter and on the physiological state of the animals. These parameters were analysed in a regression model for the requirements of nitrogen absorbed in the small intestine in relation to nitrogen balance of dry cows fed at maintenance level. The amount of total dietary nitrogen and the apparent nitrogen digestibility vary notably according to energy intake and energy concentration in dietary dry matter. On the contrary, the amount of nitrogen absorbed in the small intestine is only slightly affected by these parameters. The discrepancy between these statements lies mainly in differences in intestinal nitrogen absorption and in altered fermentation processes of the large intestine in response to changes in energy supply and ME concentration in dietary dry matter.     

Efficacy of nemadectin, a new broad-spectrum endectocide, against natural infections of canine gastrointestinal helminths

Nemadectin, a new broad-spectrum endectocide, was highly efficacious against natural infections of all the major canine gastrointestinal helminths. At single oral dosages of 0.2-0.4 mg kg-1 body weight (BW), a liquid formulation administered in gelatin capsules was 100% effective in eliminating natural infections of Toxocara canis, Toxascaris leonina, Ancylostoma caninum and Uncinaria stenocephala. Tablets (267 mg) containing 4.6% nemadectin given at a rate of 1/3 tablet per 20 kg BW (0.2 mg nemadectin kg-1) were 100% active against T. canis, A. caninum and U. stenocephala. With both formulations, an increase in the dose rate to 0.6-0.8 mg kg-1 BW resulted in 99-100% elimination of Trichuris vulpis infections. No adverse reactions were observed in any of the treated dogs.     

Regulation of Capacitation of Canine Spermatozoa during Co-culture with Heterologous Oviductal Epithelial Cells

Progress of essential steps of the capacitation is coordinated in the oviductal isthmus, where sperm are stored in close contact with the epithelium. A crucial capacitational event is the phosphorylation of sperm membrane proteins. Regulation of the tyrosine phosphorylation by the oviduct has not been examined in dog sperm yet. The aim of this work was to study the effect of dog sperm binding to porcine oviductal epithelium on capacitation‐induced cellular and molecular changes. Epithelial cells were stripped from the oviducts of post‐puberal sows and cultured for 5–7 days at 39°C and 5% CO₂ on Biomatrix‐covered Chamber slides. Sperm washed through Percoll was co‐incubated with the oviductal epithelium cell cultures in a bicarbonate Tyrode's medium. During co‐incubation, sperm membrane changes, the state of tyrosine phosphorylation and motility were determined after 3, 30, 90, 180, 240 and 360 min. Significant increases in the percentage of capacitated and dead cells were observed in unbound sperm, while bound sperm remained uncapacitated, live and motile. An increasing tyrosine phosphorylation of tail proteins in bound, unbound and control sperm suspensions and a subsequent phosphorylation of head proteins in unbound and control sperm suspensions were observed. A significant difference regarding head phosphorylation (p < 0.05) was found between sperm bound to oviductal epithelium and unbound sperm. Binding occurred mainly in sperm with non‐ phosphorylated heads, while higher proportions of phosphorylated cells were found in unbound populations. The head phosphorylation progressed significantly during incubation in unbound spermatozoa (p < 0.05); however, it was suppressed in population of sperm attached to oviductal epithelium. Significant correlations between motility parameters related to hyperactivation and tail phosphorylation were found in unbound sperm. These observations support the hypothesis that spermatozoa with non‐phosphorylated heads preferentially attach to epithelial cells. It can be concluded that tyrosine phosphorylation of head membrane proteins and capacitation are delayed in canine spermatozoa being in closed contact with oviductal epithelium.     

Incidence of Salmonellae in Captive and Wild Free‐Living Raptorial Birds in Central Spain

A total of 595 faecal samples from raptorial birds, either captive or free‐living, residing in GREFA Wildlife Hospital were bacteriologically examined using various selective media and an Automated Diagnostic Assay System for Salmonella detection. Serotype and phage type of the strains identified as Salmonella was determined. In the captive group, of the 285 samples examined, 21 (7.36%) were positive for Salmonella. Serotyping revealed that most of the individuals were infected by Salmonella serotype Havana. This result suggested that there could be a source of contamination in the Hospital although it could not be established. In the wild free‐living group, over 310 samples examined (4.19%) were positive for Salmonella. The Salmonella isolates showed a major variety of serotypes: Enteritidis, Adelaide, Brandenburg, Newport, Typhimurium, Hadar, Saintpaul and Virchow. Most of them are similar to those commonly described in isolates from human and domestic animals. These results indicate that wild birds could be involved in the dissemination of Salmonella in humans or domestic animals or vice versa.     

Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.