Enzyme immunoassays for the determination of ovine LH and FSH.

The development of competitive enzyme immunoassays for ovine plasma LH (oLH) and FSH (oFSH) is described. Standards and plasma samples were preincubated with diluted antiserum to oLH or oFSH and the reacted solution (100 micro l per well) was transferred to plates previously coated with oLH or oFSH, respectively. The second antibody used was anti-rabbit IgG horseradish peroxidase. The measuring range was 0.39-50 ng/ml for each hormone and the 50% relative binding sensitivity was 9 ng/ml for oLH. The respective value for oFSH was 3.5 or 34 ng/ml with different hormone and antibody preparations used for the assay. The enzyme immunoassays were used to determine oLH and oFSH levels in plasma from ewes of two breeds during the oestrous cycle. The assays detected the first FSH surge coincident with the LH surge, the second FSH surge about 24 h later and the periodic fluctuations of FSH concentrations during the luteal phase of the oestrous cycle. These enzyme immunoassays are an efficient and economic alternative to the established radioimmunoassays (RIA) for oLH and oFSH.

Infectivity, predilection sites, and freeze tolerance of Trichinella spp. in experimentally infected sheep.

A total of 36 sheep in groups of 4 were inoculated with 9 isolates of Trichinella and euthanized after 10 weeks. Thereafter, numbers of muscle larvae were determined in 13 different muscles/muscle groups. Muscle larvae were found in high numbers in all four sheep inoculated with T. spiralis, in lower numbers in two sheep inoculated with T. pseudospiralis (USA isolate), and in very low numbers in one sheep inoculated with T. pseudospiralis (USSR isolate) and one inoculated with T. britovi. In infections of high and moderate larval intensity, predilection sites of T. spiralis were the masseter muscles, the tongue, and the diaphragm and those of T. pseudospiralis were the masseter muscle and the neck. In low-intensity infections, muscle larvae were detected only in the diaphragm or in pooled muscle samples. For evaluation of the freeze tolerance of the different Trichinella species in sheep-muscle tissue, samples taken from the filet were stored at +5 degrees, -5 degrees, and -18 degrees C, respectively. After exposure for 1 and 4 weeks the tissue was digested and the released larvae were inoculated into mice for determination of the reproductive capacity index (RCI). Larvae of both T. spiralis and T. pseudospiralis survived freezing at -5 degrees and -18 degrees C for 4 weeks.

Effect of monensin and progesterone priming on ram-induced reproductive performance of boutsiko mountain breed ewes.

The effects of monensin and progesterone priming on reproductive performance (estrous response, lambing rate and prolificacy) of grazing Boutsiko mountain breed adult and 18-mo.-old ewes at the end of seasonal anestrus were investigated. In Experiment 1 the feed supplement with or without monensin was offered for 21 d after introduction of vasectomized rams (Day 0). Progesterone was administered to the ewes in the respective groups as a single injection at Day -3. Ewes of both age groups were assigned randomly to 1 of 4 treatments: C, C+P, C+M and C+M+P. In Experiment 2 the supplement C or M was offered from Day -26 to Day 21. The treatments consisted of C, C+P and C+M+P. Blood samples were taken 50 h after ram introduction for determination of plasma concentrations of P and insulin-like growth factor-I (IGF-I). There was a greater increase in estrous response at Days 17 to 19 and at Days 0 to 19 when supplementation was offered before rather than after ram introduction in both age groups. In the adult group ewes synchronization of estrus at Days 17 to 19 was significantly increased by administration of monensin (P<0.05) and progesterone (P<0.01) compared with the control group in the first but not the second experiment. The incidence of estrus at Days 17 to 19 or at Days 0 to 19 was highest in the adult groups treated with monensin and progesterone in both experiments. In 18-mo.-old ewes progesterone was effective in synchronizing estrus only in Experiment 2. Mean plasma IGF-I concentrations were increased by monensin treatment (P<0.05) in adult ewes that were at the periovulatory stage at blood sampling time. Correlation coefficients between IGF-I and progesterone concentrations in monensin plus progesterone group adults were -0.715 (P<0.02) and -0.516 (P<0.01), respectively across all treatments. The results suggest that monensin and progesterone priming improved reproductive performance, and the monensin-induced increase in plasma IGF-I levels at the periovulatory stage may be causally related to the ability of ovulatory follicles to develop into functional corpora lutea (CL).

Seasonal variation of gastrointestinal nematodes of sheep in the region of Joannina, Greece.

Parasitological and growth studies on two groups of naturally infected sheep, with or without anthelmintic treatment, from the age of 3 months to 2 years were carried out in the region of Joannina, Greece. A split-plot design was used so that each group, consisting of seven pure-bred Boutsiko and seven cross-bred Boutsiko with Karamaniko (F1) lambs, grazed separate parasitologically equivalent pasture plots. Faecal egg counts, pasture larval counts, plasma pepsinogen levels and live weight were recorded monthly. Infective larvae on each pasture plot increased during autumn and winter. Mean faecal egg counts for strongyle-type eggs were higher in the non-treated than the treated group and in the cross than the pure-bred sheep. Mean plasma pepsinogen levels were higher during autumn of the second year of the study. The results of the study suggest that the factors affecting the epidemiology of gastrointestinal nematodes of naturally infected sheep during grazing in the region of Joannina include anthelmintic treatment, host genotype and season, while the effectiveness of anthelmintic treatment in this study, as applied in the area, was questionable.

The effect of dietary protein levels before turnout on subsequent faecal nematode egg output of grazing sheep in the Joannina region of Greece.

The effect of dietary protein levels before turnout on the subsequent faecal egg output of gastrointestinal nematodes was examined in two genotypes of grazing sheep, with or without anthelmintic treatment. Lambs were given a low protein (LP) or a high protein (HP) diet for three months (December to March) until grazing started. Half of the animals in each diet group were given anthelmintic treatment (AT) in March and June. The four groups (LP-AT, LP-NT, HP-AT and HP-NT) each consisted of pure-bred Butsiko and cross-bred Butsiko/Karamaniko lambs and grazed separate plots that were initially parasitologically and nutritionally similar. Faecal egg counts were recorded monthly until the experiment ended in July. Mean faecal counts for strongyle-type eggs were higher in the LP-NT group than all the other groups and higher in the LP than the HP groups indicating that faecal egg counts during grazing were influenced by levels of dietary protein before turnout. This effect was intensified in the absence of anthelmintic treatment. No genotype differences were detected.

Effects of melatonin and plane of nutrition on mammary development in prepubertal Boutsiko mountain breed ewe lambs.

The effects of melatonin (implants, M or no implants, C) and plane of nutrition (high, H or low, L) on mammary development and growth hormone (GH) concentrations were investigated in prepubertal Boutsiko mountain breed ewe lambs. Eighty female lambs were assigned to each of 4 treatments: ad libitum feeding control (HC), HM, LC and LM. The rearing treatments started and ended at mean ages of 63 and 160 d, respectively. Feed restriction resulted in a mean daily gain of 70.6% of the ad libitum-fed lambs during the experimental period. Melatonin (18 mg Regulin) was administered at 68 d of age (January 10) and replaced on March 1. Blood samples were collected from 10 lambs in each treatment group at the end of the experiment for GH measurements. At a mean age of 160 d, seven lambs from each treatment group were slaughtered and the udder was removed. One udder half was trimmed and the parenchyma and fat pad portions were kept for determination of deoxyribonucleic acid (DNA) content. Melatonin did not influence mammary development parameters, while the mass of parenchyma tended to be greater in lambs on low than high nutrition planes (P<0.10). Mean mammary parenchymal weight and DNA content were 25.1 and 29.2 g and 52.5 and 58.2 mg in high and low nutrition lambs, respectively. Mean plasma GH concentrations were not affected by melatonin treatment and were higher in low than high nutrition lambs (P<0.01). There were no correlations between mean plasma GH concentrations and parenchymal DNA content, or between mean daily weight gain and parenchyma (g), in contrast to those found in a previous experiment with lambs of the same breed but greater age at slaughter. The results suggest that a period of accelerated mammary development occurs later than 140 d of age in Boutsiko mountain breed ewe lambs.

Ratios of transfer coefficients for radiocesium transport in ruminants.

A corollary of the multiple-compartment model for the transport of trace elements through animals was tested for cows, goats, and sheep. According to this corollary, for a given body "compartment" k of the animal (soft tissue, lung, liver, etc.), the ratio a(k) = f(k)/f(blood) of the transfer coefficients f, should exhibit similar values for physiologically similar animals. In order to verify this prediction, two experiments were performed at the Agricultural Research Station of Ioannina and at the facilities of Ria Pripyat in Pripyat, Ukraine. Eight animals in the first experiment and eighteen in the second were housed in individual pens and were artificially contaminated with a constant daily dose of radiocesium until equilibrium was reached. The animals were then sacrificed and transfer coefficients f(k) to twelve body "compartments" k were measured. These data were used to calculate the ratios a(k). The results were in accordance with predictions of the model and average values of a(k) were extracted for ruminants. It is concluded that these values may be employed for the prediction of animal contamination in any body compartment through the measurement of blood samples.