Molecular cloning and physiological effects of brushtail possum interleukin-1beta

Interleukin-1beta (IL-1beta) was isolated from LPS-stimulated brushtail possum alveolar macrophages using PCR primers based on conserved regions of mammalian IL-1beta. The complete cDNA was cloned by 5' and 3' rapid amplification of cDNA ends (RACE). The predicted protein of 269 amino acids shared 4346% identity with several mammalian IL-1beta proteins. Constructs were made to express the mature IL-1beta in Escherichia coli and two recombinant IL-1beta proteins, rpIL-1beta1 and rpIL-1beta2, which differed in length by four amino acids at the N-terminus, were produced. Both proteins induced a weak proliferative response in a possum thymocyte assay. Possums injected intravenously with 100 microg of rpIL-1beta1 or rpIL-1beta2 showed profound changes in body temperature and numbers of circulating leukocytes. A sharp decrease in temperature occurred within 2 h of administration followed by an elevation of temperature peaking at 24 h. The smaller rpIL-1beta1 protein had a greater effect on temperature than rpIL-1beta2. Both rpIL-1beta proteins caused a marked decrease in number of neutrophils and lymphocytes at 2-6 h after injection. At 24 h after injection, neutrophil and lymphocyte numbers were elevated 6.0-fold and 2.6-fold, respectively in the possums injected with rpIL-1beta1 and 3.9-fold and 1.5-fold, respectively in the possums injected with rpIL-1beta2. Fibrinogen levels were elevated at 24 and 72 h after injection with both proteins. In comparison, neither recombinant bovine IL-1beta (rbIL-1beta) nor PBS had significant effects on body temperature or blood haematology. The studies have shown that the two recombinant forms of IL-1beta were biologically active in possums and that the IL-1beta with four fewer amino acids at the N-terminus was the more active.     

Dynamics in the membrane organization of the mammalian sperm cell and functionality in fertilization

The capacitation process of sperm cells involves complex changes in the composition and orientation of molecules at the surface of the sperm cell. Here we focus on the lipid architecture in the sperm plasma membrane and demonstrate that the sperm plasma membrane is not static but is an extremely dynamic structure. Advanced fluoroscopic techniques enabled continuous monitoring of lipid organization in living cells and extremely rapid lipid movements were observed. The orientation of lipids in the sperm plasma membrane changed under capacitative treatments, was found to be sensitive for temperature and also changed upon binding of sperm cells to the zona pellucida. The changes in membrane properties coincided with an activation of protein kinases resulting in tyrosine phosphorylation of specific plasma membrane proteins. The detected membrane changes relate to intrinsic membrane properties such as fluidity, permeability, adhesiveness and fusibility. We think that these results may provide a physiological basis for new assays, able to discriminate between functional and non-physiological sperm cells.     

Multispecies and multiple anthelmintic resistance on cattle nematodes in a farm in Argentina: the beginning of high resistance?

There are very few resistance records on cattle nematodes. South American successive records have been increasing rapidly since the year 2000. In Argentina, increasing dissatisfaction based on the exclusive use of macrocyclic lactones has prompted the use of benzimidazoles in the Pampean region. The studied farm is located in the Argentina humid Pampas and had apparently poor results after anthelmintic treatments. Evaluation of resistance was firstly based on faecal egg reduction after treatment in November 2001 and June 2002 and was complemented with worm counts obtained by necropsy in June 2002. The study reports that the reduction of faecal egg excretion after benzimidazole (reduction 31-79%) or ivermectin treatments (76-97%) in November 2001 was insufficient and these poor results were confirmed in June 2002 (benzimidazoles (65-89%) and ivermectin (47-77%)). Several methods for evaluating faecal egg count reduction were performed and yielded different results. A new method based on the estimated faecal egg counts (corrected from the initial faecal egg counts and animal type using a general linear model) was used and gave higher flexibility in the interpretation of putative resistance to anthelmintics. From necropsy results, it could be concluded that Cooperia oncophora was resistant to avermectins and benzimidazoles and that Cooperia punctata, Ostertagia ostertagi and Haemonchus placei were resistant to benzimidazoles. This case of multispecies and multidrug resistance is probably not unique and could reflect the emergence of resistances in Argentina. This emergence is probably due to the intensive use of anthelmintics, the absence of refugia, and the frequent circulation of infected cattle.     

Keratitis due to Histoplasma spp. in a horse

A 5-year-old Holsteiner gelding from Germany was presented 2 months after a whitish discoloration of the left cornea was observed. Cytologic examination revealed intra- and extracellular globular structures, up to 4 micro m in size, consisting of a central spherical deeply basophilic body surrounded by an unstained halo. The structures were morphologically consistent with Histoplasma spp. Infection with Histoplasma organisms is not endemic in Europe. Topical use of fluconazole was successful in eliminating Histoplasma organisms within 10 days of initiation of treatment.     

Influence of grazing dormant native range or winter wheat pasture on subsequent finishing cattle performance, carcass characteristics, and ruminal metabolism

A winter grazing/feedlot performance experiment repeated over 2 yr (Exp. 1) and a metabolism experiment (Exp. 2) were conducted to evaluate effects of grazing dormant native range or irrigated winter wheat pasture on subsequent intake, feedlot performance, carcass characteristics, total-tract digestion of nutrients, and ruminal digesta kinetics in beef cattle. In Exp. 1, 30 (yr 1) or 67 (yr 2) English crossbred steers that had previously grazed native range (n = 38) or winter wheat (n = 59) for approximately 180 d were allotted randomly within previous treatment to feedlot pens (yr 1 native range = three pens [seven steers/pen], winter wheat = two pens [eight steers/pen]; yr 2 native range = three pens [eight steers/pen], winter wheat = four pens [10 or 11 steers/pen]). As expected, winter wheat steers had greater (P < 0.01) ADG while grazing than did native range steers. In contrast, feedlot ADG and gain efficiency were greater (P < 0.02) for native range steers than for winter wheat steers. Hot carcass weight, longissimus muscle area, and marbling score were greater (P < 0.01) for winter wheat steers than for native range steers. In contrast, 12th-rib fat depth (P < 0.64) and yield grade (P < 0.77) did not differ among treatments. In Exp. 2, eight ruminally cannulated steers that had previously grazed winter wheat (n = 4; initial BW = 407 +/- 12 kg) or native range (n = 4; initial BW = 293 +/- 23 kg) were used to determine intake, digesta kinetics, and total-tract digestion while being adapted to a 90% concentrate diet. The adaptation and diets used in Exp. 2 were consistent with those used in Exp. 1 and consisted of 70, 75, 80, and 85% concentrate diets, each fed for 5 d. As was similar for intact steers, restricted growth of cannulated native range steers during the winter grazing phase resulted in greater (P < 0.001) DMI (% of BW) and ADG (P < 0.04) compared with winter wheat steers. In addition, ruminal fill (P < 0.01) and total-tract OM digestibility (P < 0.02) were greater for native range than for winter wheat steers across the adaptation period. Greater digestibility by native range steers early in the finishing period might account for some of the compensatory gain response. Although greater performance was achieved by native range steers in the feedlot, grazing winter wheat before finishing resulted in fewer days on feed, increased hot carcass weight, and improved carcass merit.