Increased secretion of luteinizing hormone in prepuberal ewes treated with an antagonist of opiates

The effect of intramuscular injection of naloxone, an antagonist of opiatergic actions, on serum concentrations of luteinizing hormone (LH) and ovarian activity in prepuberal ewes was investigated. Naloxone stimulated release of LH over a period of approximately 3 h. Subsequently, serum concentrations of LH did not differ from controls. Serum concentrations of estradiol-17β were elevated 8 h after treatment with naloxone. Ewe lambs did not ovulate. Endogenous opioid peptides may be involved in the regulation of secretion of LH in immature female sheep.     

Effects of estradiol-17β, para-chlorophenylalanine and quipazine on tonic and LHRH-induced secretion of luteinizing hormone in ovariectomized ewes

Changes in metabolism of serotonin (5-HT) might mediate the reduced tonic luteinizing hormone (LH) and increased pituitary responsiveness to luteinizing hormone releasing hormone (LHRH) caused by estradiol-17β (estradiol). Two experiments were conducted to determine effects of estradiol, para-chlorophenylalanine (PCPA), an inhibitor of synthesis of 5-HT, and quipazine, an agonist of 5-HT, on tonic and LHRH-induced secretion of LH in ovariectomized ewes during the summer. Tonic levels of LH were reduced, the interval from LHRH to peak of the induced surge was longer and the magnitude of release of LH was greater in ovariectomized ewes treated with estradiol than in controls. Neither PCPA nor quipazine affected tonic secretion of LH. In ovariectomized ewes not receiving estradiol, PCPA and quipazine increased the magnitude of the LHRH-induced release of LH. However, PCPA reduced pituitary sensitivity to LHRH when administered concomitantly with estradiol; treatment with quipazine attenuated this effect of PCPA. The interval to the peak of the induced surge of LH was not affected by PCPA or quipazine in estradiol-treated or control ovariectomized ewes. Based on these results it appears that 5-HT mediates or is required for estradiol to increase pituitary responsiveness to LHRH.     

Progesterone and the surge release of gonadotropins in the ewe

On day 12 of an estrous cycle, 4 groups of ewes were treated with either blank (no steroid) or 3 different sizes of progesterone-containing rubber implants to study the effect of maintained progesterone levels on preovulatory events. Following luteolysis progesterone levels were 0.55 ± 0.13 ng/ml in control ewes and 0.62 ± 0.10 ng/ml, 0.99 ± 0.09 ng/ml and 1.85 ± 0.04 ng/ml in the 3 groups of progesterone-treated ewes. Preovulatory surges of LH and FSH occurred in $\text{5}\text{5}\text{,}\text{5}\text{5}\text{;}\text{4}\text{5}\text{,}\text{3}\text{5}\text{;}\text{4}\text{5}\text{,}\text{3}\text{5}$ and $\text{0}\text{5}\text{,}\text{0}\text{5}$ ewes in these 4 groups respectively. Eleven days after implant insertion, all ewes responded to a GnRH challenge. The height of all FSH peaks was depressed by progesterone treatment (P<0.05). Three groups of ewes were ovariectomized at day 6 of a cycle and treated with estradiol-17β and progesterone-containing implants. After 8 days of treatment, progesterone implants were removed in sections to give 3 different rates of decline in serum progesterone levels. The gonadotropin surges occurring following progesterone removal were delayed by the slower rates of progesterone decline. Another group of ewes was treated as in experiment 2, but the progesterone implants were all removed together after 8 days. Subsequent replacement of progesterone implants, after 12 or 18 hours, blocked the gonadotropin surges in all, or 2 of 6 ewes respectively. When replaced after 24 hours, implants producing low progesterone (1.38 ± 0.22 ng/ml) did not block gonadotropin surges, but in 2 of 5 ewes high progesterone did (2.87 ± 0.22 ng/ml). Removal of progesterone implants, 12 or 24 hr after replacement, produced secondary gonadotropin surges of smaller magnitude than the initial peaks (P<0.01).     

Regional differences in concentrations of LHRH receptors,LH and FSH in the bovine adenohypophysis

Pituitaries from intact luteal phase (INT) and ovariectomized (OVX) cows were collected at slaughter to determine whether differences exist among regions of the bovine adenohypophysis in the concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and receptors for luteinizing hormone releasing hormone (LHRH). Each adenohypophysis was divided into three paired regions (anterior, AT; medial, M; posterior, PT) by first making a midsagittal cut followed by two transverse cuts of approximately equal size. Values for all variables were similar between paired regions. Mean LHRH receptor, LH and FSH concentrations were greater in OVX than INT adenohypophyseal regions. Receptor and gonadotropin concentrations differed among all three regions and were greatest in the AT, intermediate in the M and lowest in the PT regions of the adenohypophysis. There were significant correlations between LHRH receptor concentrations and concentrations of LH and FSH among the three adenohypophyseal regions for both INT and OVX cows. Therefore, to accurately characterize LHRH receptors from the bovine adenohypophysis, a midsagittal-half of the gland should be used for analysis.     

In vitro release of LHRH from the bovine pituitary stalk-median eminence: Alterations during the postpartum period

Superfusion techniques were developed to investigate in vitro release of luteinizing hormone releasing hormone (LHRH) from the bovine pituitary stalk-median eminence (SME) obtained from suckled beef cows during the postpartum period. Mature beef cows were allotted at random to be slaughtered at either day 5 (n=5), 10 (n=6), 20 (n=6) or 30 (n=5) after calving. After isolation from the brain, the pituitary SME was divided mid-sagittally with one-half of the tissue placed in a vessel containing ice-cold superfusion medium and transported to the laboratory for superfusion. Krebs-Ringer phosphate solution supplemented with BSA, glucose and bacitracin served as the superfusion medium. A 10-min pulse of high K⁺ Krebs-Ringer phosphate medium was passed through the superfusion chamber at two different times to provoke membrane depolarization causing release of LHRH. Treatment of SME tissue with high K⁺ media increased LHRH release over basal pre-K⁺ levels attesting to the viability of the preparation. The maximum K⁺-induced peak release of LHRH at the first K⁺ pulse was greater on day 5 than day 20. These results indicate that: Dour superfusion system can be used to evaluate releasable pools of LHRH from the bovine pituitary SME, and 2) in vitro release of LHRH in response to K⁺ depolarization is reduced by day 20 postpartum possibly indicating increases in in vivo release between day 5 and day 20.     

The possible effects of gastro-intestinal nematodes upon intestinal enzymes of calves

Observations were made on the effect of combined infections of gastro-intestinal nematodes on small intestinal disaccharidases and alkaline and acid phosphatases in 4–5-month old Friesian calves. The tests were carried out 3 weeks following exposure of these calves to infected pasture in a wet tropical environment.Sucrase activity, at a very low level, was detected in all parts of the small intestine of infected and uninfected calves. There was no significant difference in maltase, trehalase, cellobiase, sucrase, lactase, acid β-galactosidase, acid and alkaline phosphatase activities between uninfected control calves and all infected calves in which Cooperia species predominated. In heavily infected calves with worm burdens greater than 20 000 Cooperia species significantly elevated (P < 0.05) duodenal maltase and acid phosphatase levels were recorded.     

The effects of different maternal dietary iodine concentrations on Japanese quail II. Thyroid function in embryos and hatchlingsi

Embryos (day I4 of the 16.5 day incubation period) and I day old chicks of Japanese quail (Coturnix japonica) were used to study the effects of egg I on thyroid development. The maternal diets were supplemented with 0–1200 μg I/kg of purified diet. Thyroid function, of embryos and chicks (before feeding), adapted to a wide range of egg I availability.We measured body weights, thyroid weights, thyroidal ¹²⁵I uptakes, stable I concentrations in thyroid glands and triiodothyronine (T₃) and thyroxine (T₄) concentrations in the serum.With low egg I (maternal diets with <50 μg I/kg) embryonic and chick thyroid glands were hypertrophied, thyroidal I content was very low and thyroidal radioiodine uptakes were high. Serum hormone concentrations were not significantly different from those of embryos and chicks from eggs with higher I contents. Thyroid weights were lowest in young from eggs of hens on the 150 μg I/kg diet, and the other variables studied also indicated this maternal diet provided sufficient egg I for the thyroid function of embryos and hatchlings.With high egg I (maternal diets with 300–1200 μg I/kg) thyroidal radioiodine uptakes were low but thyroidal I incorporation was related to egg I content. Serum hormone concentrations were not significantly altered by high I availability except that serum T₄ concentrations differed in a comparison of the extremes (0 vs 1200 μg I/kg in the maternal diet). As expected, from the consistency of serum hormone concentrations, embryonic growth was not affected by the I content of the eggs.In agreement with earlier studies on quail hens, our results indicate that feed supplementation of 150 μg I/kg is sufficient to meet needs without requiring alterations in thyroid function to maintain serum hormone concentrations. Developing thyroid glands adapt effectively to egg I concentrations of 0.2 to 4.1 μg I/g yolk which result from maternal diets with 0–1200 μg I/kg.     

The effects of different maternal dietary iodine concentrations on Japanese quail I. Thyroid status of hens

Japanese quail Coturnix japonica were used as a model system for studying the effects of different levels of dietary iodine (I) supplementation (0–1200 μg I/kg of purified diet) on thyroid function and egg-laying in adult galliform birds. Quail have the ability to adapt their thyroid function to a wide range of dietary I intakes.We measured thyroidal ¹²⁵I uptakes, stable I concentrations in serum and thyroid glands, and triiodothyronine (T₃) and thyroxine (T₄) concentrations in thyroid glands and serum. Body weights of the hens as well as egg production, egg hatchability, and yolk I content were monitored regularly throughout the 56 week study.With low dietary I (0–50 μg I/kg) and consequent low serum I, there is marked stimulation of thyroidal I uptake. Thyroidal stores of I and T₄ are reduced, but thyroidal T₃ and serum concentrations of both T₃ and T₄ are maintained.On dietary I regimes of 150–1200 gg I/kg, thyroidal I stores are regulated at essentially the same level despite a linear relationship between dietary I and serum I concentrations. Thyroidal hormone content and serum hormone concentrations also do not differ between dietary I regimes with >I50 μg I/kg.Egg production and egg hatchability are maintained on all dietary I intakes, a result consistent with the maintenance of serum T₃ and T₄ concentrations independent of dietary and serum I. In contrast to the thyroid, the ovaries do not appear able to regulate I in relation to I availability, and egg yolk I is proportional to serum I concentrations.These studies suggest that feed supplementation of 150 μg I/kg is sufficient to meet the needs of Japanese quail hens without requiring alterations in thyroid function. Quail adapt effectively to dietary I from 0–1200 μg I/kg; maintenance of thyroid status and egg production are favored while egg I stores vary with I availability.     

Branched Mycobacterium-like organisms from swine lymph nodes

Branched, weakly acid fast organisms were isolated from 5 of 112 caseous lymph nodes derived from slaughter-house pigs. In two cases they were associated with typical Mycobacterium avium strains. The bacteria differed from all other mycobacteria known at present. Limited experiments in pigs with one strain failed to disclose a possible pathogenic role in the host species although this strain was pathogenic for mice. Tests with this and three other strains showed that all four were apathogenic for chickens.     

Analysis and in vivo assay of B. abortus agglutinating and non-agglutinating antibodies

Studies were made of physicochemical and immunochemical characteristics of Brucella abortus agglutinating and non-agglutinating antibodies in the sera of cattle repeatedly injected with living B. abortus (Strain 1119). Both agglutinating and non-agglutinating antibody were shown to be IgG1, and by immunodiffusion against rabbit anti-cattle gamma-globulin, agglutinating antibody gave a precipitation line of identity with that given by non-agglutinating antibody. Whilst agglutinating antibody increased clearance of antigen from the blood of passively protected mice, non-agglutinating antibody did not enhance clearance. Determination of the spleen infection index in mice pre-treated with agglutinating and non-agglutinating antibody showed that in animals passively immunized with non-agglutinating antibody the number of living (infecting) bacteria was approximately 4 times higher than in the case of agglutinating antibody. The possible potentiation of chronic B. abortus infection by non-agglutinating antibody is discussed.     

Dairy farm wells in southeastern Minnesota: The relation of new water sources (new wells) to milk and milk fat production

Milk and milk fat production per cow were studied on 17 Fillmore County, Minnesota, Dairy Herd Improvement Association dairies before and after the source of drinking water for cows was changed by the drilling of a new well. New wells were in deeper aquifers which were more protected from waterborne pollutants; the former wells were in shallow, easily polluted aquifers. Seventeen other dairies with swallow wells served as controls, matched for pre-well milk production per cow, dairy location, and breed of cow. The changes in milk and milk fat production per cow after the drillingof a new well were not statistically significant.     

Dairy farm wells in Southeastern Minnesota: The relation of water source to milk and milk fat production

Some farmers in Fillmore County, Minnesota, U.S.A., considered that contaminated well water was associated with lowered milk production in their dairy herds. Wells in shallow limestone-dolomite aquifers were easily contaminated by surface water. Fifty Dairy Herd Improvement Association dairies were classified by whether well water originated in deep, protected aquifers or in shallow, easily contaminated aquifers. No statistically significant differences in milk fat production were demonstrated between the two groups before or after the use of linear multiple regression models to control for differences in management practices.     

Dairy farm wells in Southeastern Minnesota: The relation of water source to calf mortality rate

Some farmers in Fillmore County, Minnesota, U.S.A., considered that contaminated well water was associated with lowered milk production in their dairy herds. Wells in shallow limestone-dolomite aquifers were easily contaminated by surface water. Fifty Dairy Herd Improvement Association dairies were classified by whether well water originated in deep, protected aquifers or in shallow, easily contaminated aquifers. No statistically significant differences in milk fat production were demonstrated between the two groups before or after the use of linear multiple regression models to control for differences in management practices.     

Growth hormone secretion in chicks during dietary calcium deficiency

The role of calcium in the secretion of growth hormone (GH) has been examined in vivo in immature domestic fowl. Chicks reared on a low calcium (0.16% calcium in the feed) diet showed a reduced growth rate, compared with those on a normal (0.86%) calcium diet and had lower basal levels of plasma GH 1, 5, 10 and 15 d after calcium deprivation, but not after 20, 25, 30 or 35 d of calcium deficiency. No consistent changes in the concentration of immunoreactive somatomedin C were observed during calcium restriction. In both the control and low calcium fed birds the plasma concentrations of GH were elevated by the intravenous administration of human pancreatic GH releasing factor (hpGRF) and by thyrotropin releasing hormone (TRH). The GH responses to these provocative stimuli were not reduced in magnitude by calcium deficiency. It is suggested, therefore, that the effect of calcium deprivation on the secretion of GH is mediated via the reduced release of stimulatory hypophysiotrophic factors from the hypothalamus.

Pharmacological alterations in calcium status also suggest that calcium deprivation inhibits GH secretion. Plasma concentrations of GH were acutely depressed in young chicks following the administration of a calcium chelator (ethylene diaminotetraacetic acid) a calcium channel blocker (verapamil) and after calmodulin inhibition (by chlordiazapine and trifluorpenazine administration). These data therefore demonstrate the importance of calcium in the stimulation of GH secretion in the domestic fowl.     

Comparison of mycoplasma ovipneumoniae isolates using bacterial restriction endonuclease DNA analysis and SDS-PAGE

Sixteen isolates of Mycoplasma ovipneumoniae recovered from the nasal tract or lungs of sheep from different flocks in New Zealand were examined by bacterial restriction endonuclease DNA analysis (BRENDA) using EcoR1 and by sodium dodecyl sulphate-polycrylamide gel electrophoresis (SDS-PAGE). All isolates gave BRENDA patterns which differed entirely from one another. Following 20 serial passages (corresponding to approximately 67 generations) of an isolate, no charge was detected in the BRENDA pattern.

When eight isolates were examined by SDS-PAGE most bands were common but, nevertheless, each isolate was unique in the sense that they differed from one another in one or more bands. The marked heterogeneity of patterns observed when strains of M. ovipneumoniae are compared by BRENDA, together with the stability of such patterns over many generations, will enable this approach to be used to study the epidemiology of individual strains of M. ovipneumoniae with a flock.     

Induction of ovulation in anestrous mares with pulsatile administration of gonadotropin-releasing hormone

Four seasonally anestrous mares (Standardbred), housed under a nonstimulatory photoperiod of 8 hours light:16 hours dark, were administered gonadotropin-releasing hormone (GnRH) in a pulsatile pattern (50 or 250 micrograms of GnRH/hour) for 8 to 18 days during February and March 1985. Treatment with GnRH, irrespective of dose or month, induced an increase in serum luteinizing hormone from a mean pretreatment value typical of anestrus (0.58 +/- 0.02 ng/ml +/- SE) to 10.84 +/- 1.27 ng/ml on day 8 of GnRH treatment. Ovulation in the 4 mares occurred 8.8 +/- 0.7 days after the initiation of pulsatile GnRH administration. In each instance, ovulation was followed by a functional corpus luteum, as indicated by a luteal phase (defined as the number of days on which serum levels of progesterone were greater than 1.0 ng/ml) which lasted 14.5 +/- 0.6 days. These results indicate that infusion of GnRH in a pulsatile pattern is effective in inducing follicular development and ovulation in anestrous mares in the absence of a stimulatory photoperiod.     

Colonisation of the respiratory tract of lambs by strains of Mycoplasma ovipneumoniae

The age and time of year when colonisation of the nasal cavity of lambs by Mycoplasma ovipneumoniae occurs; the persistence of the organism, and its prevalence in the lungs at slaughter were examined in 2 flocks of sheep in New Zealand. No colonisation had occurred at the time of weaning at 6–7 weeks, but M. ovipneumoniae was recovered from most lambs on at least one occasion before they were slaughtered when about 8 months old. In most cases, colonisation of the nasal cavity by M. ovipneumoniae was a transient phenomenon. At slaughter M. ovipneumoniae was recovered from the lungs of 89% of the lambs of one flock and 80% of the other flock.

Bacterial restriction endonuclease DNA analysis (BRENDA) of 34 nasal isolates from one flock showed that it was possible to identify 7 “groups” each with markedly different BRENDA patterns. Lambs initially colonised by one strain, often lost that strain, and if recolonisation occurred it was with a different strain.

M. ovipneumoniae was recovered at slaughter from the lungs of most lambs, both normal and pneumonic. The isolates from one flock were examined by BRENDA, and approximately 90% of them gave similar or identical patterns. The predominant strain isolated from the lungs had been recovered from the nasal cavity of many of the lambs about 3 weeks earlier. This suggests that the nasal and lung isolates do not represent independent populations. However, nasal strains may differ in their ability to colonise the lungs.     

Kinetics of infection with Theileria parva (East Coast fever) in the central lymph of cattle

During the course of a lethal infection with Theileria parva in susceptible cattle, the dissemination of the parasite was examined in central lymph efferent from superficial lymph nodes in the thoracic duct. From the regional node, lymphocytes containing macroschizonts of T. parva were detected in efferent lymph 8 days after challenge where their appearance coincided with a dramatic increase in the output of lymphoblasts. The number of infected cells reached a maximum around Day 14, when 60-65% of efferent lymphocytes were parasitized. A severe reduction in the total cell output occurred after Day 14, at the time when widespread lymphocytosis was observed in the parent lymph node. A similar pattern of cellular kinetics was observed in the thoracic duct and in lymph efferent from lymph nodes distant from the site of challenge, although in the latter, the parasitosis reached only 10% of total cells. There was no selective depletion of parasitized cells from central lymph during the third week of infection, although the comparative parasitosis between lymph and lymph node cells indicated that infected cells entered central lymph less readily during this period. Macroschizonts appeared in cultures of lymphatic lymphocytes sampled between 5-9 days after challenge. These results, together with the failure of ablation of the regional lymph node 2, 3 or 5 days after challenge to delay the onset of the disease, indicated that dissemination of the infection from the site of challenge occurred within the first 2-3 days after the inoculation T. parva.     

Dirofilariasis in wild canids from the Gulf coastal prairies of Texas and Louisiana,U.S.A.

Heartworms, Dirofilaria immitis, were recovered from 17 out of 24 (71%) coyotes, Canis latrans, 38 out of 46 (83%) coyote × red wolf hybrids and all of 8 (100%) red wolves, Canis rufus gregoryi, collected from the Gulf coastal prairies of southeast Texas and southwest Louisiana. Intensities of infection ranged from 1 to 176 ($\text{x}\text{=25}$) worms per host. There was a significantly (P<0.05) higher intensity of infection in red wolves. Prevalence of heartworms increased significantly with increasing age. There were no significant differences between coyotes, hybrids and red wolves or between different host sexes in terms of prevalence. The female to male ratio of heartworms was close to unity (1.2:1) and was not correlated with worm burdens. Nematodes were primarily localized in the right heart, frequently extending into the pulmonary artery and the pulmonary arterial tree in the lungs. In 13 instances, 1–4 adult heartworms were recovered from the venae cavae. Pathological responses in the right heart were variable, depending on the intensity of infection. In severe infections, there were small areas of infarctive necrosis with mild to severe interstitial edema in the myocardium. Lesions in the pulmonary artery and pulmonary arterial trunk varied from mild focal hyperplastic intimal changes to extensive exudative villous endarteritis. The latter was characterized by a hyperplastic collagenous stroma with numerous histiocytes, plasma cells and eosinophils. Lung pathology varied from patchy to extensive areas of congestion, edema, hemorrhage, interstitial pneumonitis and infarction. In cases with heartworms in the venae cavae, hepatic changes were minimal and associated with liver changes such as passive congestion and centrolobular necrosis seen in cases without adult worms in the venae cavae. In heavily infected animals, hemosiderosis of the liver, spleen and kidneys was pronounced. A microfilaremia was noted in 46% of the infected wild canids. Microfilariae were observed in tissue sections of the myocardium, lungs, liver, kidney, spleen, lymph nodes, pancreas and appendix. Wild canids from this area are regarded as natural definitive hosts and primary reservoirs for heartworms and it appears that this infection is an important factor in the morbidity and mortality of these hosts.