LHRH Fusion Protein Immunization Alters Testicular Development, Ultrasonographic and Histological Appearance of Ram Testis

This study was designed to evaluate the effectiveness of a luteinizing hormone releasing hormone (LHRH) fusion protein immunization on reproductive traits in ram lambs including the changes in histology and ultrasonographic appearance of testis. Thirty native ram lambs at 19 weeks of age were divided into control (C, n = 10), immunization (I, n = 10) and castration (E, n = 10) groups. Animals in immunization group were immunized against LHRH using ovalbumin‐LHRH‐7 (OL) protein generated by recombinant DNA technology as a primary and a booster injection at 19 and 23 weeks of age respectively. Animals were bled via jugular venepuncture at 2‐week intervals to determine LHRH antibody and testosterone concentrations. Bi‐weekly ultrasonographic examination of the testes was performed to determine the changes in ultrasonographic appearance as the age increased. Biopsied testicular tissues taken at 19, 29 and 41 weeks age were also evaluated. At 41 weeks of age, animals were slaughtered. Semen and epididymis were evaluated for the presence of sperm cells. Testicular development and sperm production were suppressed in the immunized animals. Semineferous tubule diameters decreased, basal membrane of the tubule was thickened and hyalinized in immunized ram lambs. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 19 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging. Nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in ram lambs and has a potential to be used as an alternative to physical castration. Further research studies should be conducted to help assess reproductive status of testes from ultrasound images.     

Effect of fishmeal replacement with spray-dried animal plasma and colistin on intestinal structure,intestinal microbiology,and performance of weanling pigs challenged with Escherichia coli K99

We evaluated spray-dried animal plasma (SDAP) as an alternative to antimicrobial medication with colistin sulfate in weanling pigs challenged with Escherichia coli K99. Forty-eight piglets weaned at 24 d of age were distributed into 12 pens, and each pen was assigned to one of four dietary treatments. All the piglets were given an oral dose of 5 x 10(7) cfu of E. coli K99 at weaning. The dietary treatments followed a factorial arrangement with two levels of SDAP (0 and 7%) and two levels of colistin (0 and 300 mg/kg of diet). The ADG and ADFI were measured on d 7 and 14 of trial. Three piglets from each treatment were killed on d 7 and 14 to remove the small intestine, and to obtain ileal and cecal digestive contents. The inclusion of SDAP improved ADG by 68 g (P < 0.05) and ADFI by 41 g (P < 0.10) in wk 1 of trial. During wk 2, SDAP improved ADG by 41 g (P < 0.10) and gain:feed ratio (G:F) by 25% (P < 0.01). On the other hand, whereas colistin had no effect on performance in wk 1, it improved ADG by 102 g (P < 0.01), ADFI by 62 g (P < 0.01), and G:F by 26% (P < 0.01) in wk 2. Over the 14 d of the trial, ADG was improved by 54 (P < 0.05) and 75 g (P < 0.05), and G:F was improved by 35 (P < 0.05) and 32% (P < 0.05) due to SDAP and colistin, respectively. There was interaction between colistin and SDAP for ADFI in wk 2 and between d 0 to 14 (P < 0.05), which indicates that their effects were not additive. The use of colistin was advantageous in the maintenance of the integrity of the intestinal mucosa of the pigs, as suggested by a small intestine that was 93 g heavier (P < 0.10) and with the tallest villi 106 microm longer (P < 0.10) than in pigs without colistin. The inclusion of SDAP in the diet favored the growth of lactobacilli in the ileum (P < 0.10) and the cecum (P < 0.05), whereas colistin reduced the number of enterococci in the cecum (P < 0.05) and of Escherichia coli both in the ileum and the cecum (P < 0.001). These results suggest that SDAP may be an alternative to medicated feed with antibiotics since it provided a level of protection against an experimental challenge with E. coli K99 similar to that obtained with colistin, an antibiotic of proven efficacy. The current situation in which the use of antimicrobials in animal feeding is being questioned should encourage further investigation into the use of SDAP as a means of preventing disease in pigs at weaning.     

Relative contribution of colostrum from Maedi-Visna virus (MVV) infected ewes to MVV-seroprevalence in lambs

Maedi-visna virus (MVV) seroprevalence associated with consumption of colostrum from seropositive ewes was investigated in 276 housed lambs from birth to 300 days-old. At birth, lambs were allocated to five experimental groups according to the maternal MVV-serological status, source and mode of feeding colostrum (bovine or ovine and bottle fed or suckled from the dam) and type of horizontal MVV-exposure (raised with the dam or separately with other lambs). The risk of being seropositive at 300 days-old was associated with feeding ovine colostrum from seropositive ewes and increased with intake of bottle-fed ovine colostrum and was higher in lambs separated from their dams and raised with other experimental lambs compared to lambs raised with their dams. Approximately 75-87% of ELISA-positive results in lambs that had ovine colostrum was attributable to colostrum itself. However, approximately only 16% of naturally raised and 29-61% of bottle-fed ovine colostrum lambs were ELISA-positive as a result feeding ovine colostrum. These results confirm that ovine colostrum from seropositive ewes can be a major source of MVV but its overall contribution to seroprevalence in natural conditions is relatively low, and shows that horizontal MVV transmission can be an important source of infection in new-born lambs.     

Comparison of immunohistochemistry and two rapid tests for detection of abnormal prion protein in different brain regions of sheep with typical scrapie.

One of the "gold standard" techniques for postmortem confirmation of scrapie diagnosis in sheep and goats is immunohistochemical examination of brain tissue. Active surveillance for scrapie is mainly performed by rapid diagnostic tests on the basis of postmortem immunochemical detection of prion protein (PrP) in the obex tissue. The aim of this study was to determine the performance of 2 rapid tests, Prionics-Check LIA (a chemiluminescence sandwich enzyme-linked immunosorbent assay) and Prionics-Check Western blot for scrapie diagnosis when applied to brain areas other than the obex, in comparison with the recognized immunohistochemistry. Prion protein was detected in the obex, cervical spinal cord, and thalamus from all the scrapie-positive sheep by the 3 tests. Western blot and LIA were negative in other areas of the brain, although weak immunohistochemical staining was detected. The results show that the 2 rapid tests studied may detect PrP in brain areas other than the obex, although with a lower sensitivity than immunohistochemistry when there is minimal PrP deposition.     

Evaluation of spray dried animal plasma and calcium formate as alternatives to colistin in piglets experimentally infected with Escherichia coli K99

Two trials were conducted to evaluate spray dried animal plasma and calcium formate as alternatives to preventive medication with colistin in piglets experimentally challenged with Escherichia coli K99. Two groups of newly weaned pigs were offered four treatments consisting of: Negative Control (NC); spray dried animal plasma (SDAP); calcium formate (CF) and colistin (COL). All animals were experimentally challenged with a single oral dose of E. coli K99. Their performance was recorded and, in the second trial, 6 piglets from each treatment were killed to obtain samples of jejunal mucosa for histological measurements and digesta from the ileum and the caecum for microbiological determinations. SDAP improved weight gain (P < 0.05) and feed intake (P < 0.06) in the first two weeks after weaning of trial 1, and a similar response, although not statistically significant, was found in trial 2. Colistin also resulted in a numerical improvement in performance, but calcium formate did not. No clear effects on mucosal histology were observed and only colistin had a significant effect on the microbiological composition of digesta.     

Luteolytic Effect of Prostaglandin F2α on Bovine Corpus Luteum Depends on Cell Composition and Contact

Prostaglandin F_2α (PGF_2α) is a main luteolytic factor in vivo; however, its direct luteolytic influence on steroidogenic cells of bovine corpus luteum (CL) is controversial and not fully understood. The aim of the study was to clarify PGF_2α action on bovine CL in different in vivo and in vitro conditions and to examine whether the contact among all main types of CL cells is necessary for luteolytic PGF_2α action. In experiment 1, the bovine CL (day 15 of the oestrous cycle) was perfused using in vivo microdialysis system with dinoprost (an analogue of PGF_2α) for 0.5 h. Dinoprost caused a short‐time increase in progesterone (P4), whose concentration decreased thereafter (at 6‐, 10‐, 12‐ and 24‐h after treatment). In experiment 2, the direct effect of PGF_2α on P4 accumulation in CL steroidogenic cells cultured in monolayer (day 15 of the cycle) was determined. PGF_2α after 24 h of incubation increased P4 accumulation in steroidogenic CL cells. In experiment 3 steroidogenic, endothelial CL and immune cells (day 15 of the cycle) were incubated with PGF_2α in cocultures for 24 h in glass tubes and the levels of P4, stable metabolites of nitric oxide (NO) and leukotriene (LT) C₄ were determined. Although PGF_2α treatment increased P4 secretion in homogeneous steroidogenic CL cell culture, the decrease in P4 secretion in cocultures of all types of CL cells was observed. The secretion of NO and LTC₄ increased after the treatment of PGF_2α both in pure cultures of CL cells and in cocultures. The interactions between endothelial and immune cells with steroidogenic CL cells are needed for luteolytic PGF_2α action within the bovine CL. Our results indicate that the cell coculture model, including the main types of CL cells, is the most approximate to study PGF_2α role in vitro.