The macroscopic and scanning electron-microscopic appearance and microhardness of the enamel, and the related histological changes in the enamel organ of erupting sheep incisors resulting from a prolonged low daily dose of fluoride.

The hypothesis that diffuse opacities in enamel result from a chronic, mild disturbance to ameloblast activities was tested using fluoride. Three sheep (HF) were dosed orally with 0.5, and 3 (LF) with 0.2 mg fluoride/kg body weight daily for 6 months. A control sheep (C) received no additional fluoride. The 7 sheep were killed at or close to the time of emergence of their permanent central incisors. One tooth from each sheep was sectioned longitudinally. The enamel related to the secretory (S) and maturation (M) phases of ameloblast activity at the start of fluoride dosing was determined from a tetracycline marker. The pattern of mineralization of the outer 150 micron of the cut labial enamel was assessed using microhardness testing. The SEM appearance of the acid-etched outer enamel was compared in S and M zones in 5 teeth. The enamel of the C tooth was translucent. Diffuse opacities, similar in appearance to human fluorosis, were present in all fluoride-treated teeth. Hardness values in the outer 70 micron of the enamel decreased as the fluoride dose increased and, in the HF teeth, were lower in the S zone than in the M zone. Fluoride given during the M phase induced a surface hypomineralization which increased in degree and depth when fluoride was also given during the S phase. The SEM appearance of M and S enamel was similar in 2 LF and 1 HF teeth but, in the other HF tooth, S enamel but not M enamel had a disordered prism structure and loosely-packed crystals in an abnormal organic matrix. Histological examination revealed that ameloblasts remained in only 4 of the 7 teeth and that their regression and the formation of the cementum adjacent only to the labial enamel were progressing abnormally.

Gingivitis around the deciduous teeth of sheep.

Two-hundred and eight sheep from two farms were killed mostly in age-matched groups of 6 between the ages of 6 and 141 weeks. Sheep from one farm (W) had excessive tooth wear whilst sheep from the other (G) had little. The four deciduous incisor teeth and the third premolar from one side of the mandible of each sheep were examined by light microscopy. The inflammatory reaction in the gingiva around the sulcus and sulcular epithelial proliferation were subjectively assessed. Sheep from both farms initially showed a neutrophilic reaction. From approximately 25 to 100 weeks of age, sheep from farm G had a more extensive reaction than those from farm W involving neutrophils and mononuclear cells. The latter showed a more frequent tendency to spread on farm G than on farm W. The length of the sulcus was also more often greater on farm G. In contrast, the proliferation of the sulcular epithelium in both width and length was not greater on farm G. It is suggested that the severity of the gingivitis was arrested at a less severe stage on farm W than on farm G and that the pathogenesis of sulcular epithelial proliferation may different from that of gingival inflammation.

The macroscopic appearance and associated histological changes in the enamel organ of hypoplastic lesions of sheep incisor teeth resulting from induced parasitism.

Twenty-one penned female sheep were infected with a high dose of nematodes (200,000 Trichostrongylus vitrinus and 25,000 Ostertagia circumcincta) during first incisor development when aged 7-8 months. Three sheep were infected with 200,000 T. vitrinus only; 10 sheep were untreated. Anorexia, lassitude and severe diarrhoea were seen in 14 of the infected sheep after 21-26 days. Anthelmintic was given to sheep showing undue distress to end the infection and to all remaining sheep on day 33. Animals were killed at 3 stages: during the infection, at intervals during later tooth development, and at the time of eruption of the incisor teeth, 7-8 months after the parasites were given. Hypoplasia of the labial enamel of 15 out of 19 teeth from sheep killed after recovery from the infection was classified according to the extent and depth as pits, grooves or larger areas of missing enamel with ledge-formation cervically. The amount of missing enamel related to the severity of the systemic disturbance which affected the secretory ameloblasts. During the period of active infection, some secretory ameloblasts showed vacuolization or were shorter than in controls; some cells were no longer adjacent to the organic matrix; some continued to function but abnormally and later became separated from the retained organic matrix by connective tissue. Cervically-placed ledges resulted either from the recovery and resumption of activity of some ameloblasts or from differentiation of new secretory cells after the infection ended.

Development, growth and eruption of permanent incisor teeth in Romney sheep.

The development of permanent incisor teeth of sheep was followed from the age of six weeks to 116 weeks. The teeth were observed through the bud, cap and bell stages, and the development of enamel and dentine. At the same time that amelogenesis started, the papillary layer developed from the intermediate layer and the outer dental epithelium began to form the cell rests of Malassez. The ameloblasts went through secretory, transitional and maturational phases, and in due course disappeared, after which cementum was laid down over the enamel surface. Initially the connective tissue of the follicle surrounding the growing tooth was very loose, but was replaced from the crown end as the tooth developed, by more dense connective tissue. Around the time of eruption, melanocytes were found on the lingual surface of the coronal end of the follicle. The root was formed under the influence of Hertwig's epithelium, and the cell rests of Malassez appeared to continue to develop from the outer enamel epithelium. Odontoblasts, which arose from the pulp connective tissue, were initially cuboidal but became fusiform as the tooth matured. At the time of eruption, amelogenesis was almost complete and cementum covered the majority of the tooth surface. The cell rests and melanocytes of the tootth follicle appeared to migrate with the tooth as it erupted. The deciduous tooth was resorbed from the lingual surface. The junctional epithelium of the erupted permanent tooth may have arisen from the cell rests, remnants of the dental lamina or gingival epithelium, depending on the presence or absence of inflammation.

Erosion of the non-occlusal surfaces of sheeps' deciduous teeth.

Major erosion of the labial or lingual surfaces was seen in histological sections of deciduous teeth from three of 208 sheep from two farms. In one, the erosion extended through both the enamel and the dentine and thus exposed the pulp cavity which had developed pulpitis. In the other cases the erosion affected the cementum and dentine of the tooth root, and did not penetrate as far as the pulp cavity. Dentine proliferation occurred on the adjacent pulpal surface, however, although pulpitis was not evident.

The pathogenesis of excessive wear in the permanent teeth of sheep.

The two central pairs of permanent incisor teeth (l1P, I2P) were compared post mortem in sheep from two flocks, one of which showed excessive wear. Sheep were usually killed in groups of six. Excessive wear occurred during the winter and spring after the eruption of l1P. When l1P erupted at an early age it was also subject to that season;s wear but when it erupted at a later age, it was subject to less wear according to the amount of time it was in apposition with the dental pad during the high wear season. A small amount of compensatory growth at the root of the tooth occurred in some sheep when cementum was deposited on the apex. Apparent eruption of the crown of the tooth was actually due to gum recession. Normal reparative dentine was laid down in the pulp cavity of the crown of the tooth in anticipation of wear, but where wear was excessive unstructured dentine was deposited. In two cases the rate of wear was so great that the reparative dentine could not keep up and infection was able to gain access to the pulp cavity. There was no evidence of difference in the rate of wear of the molar teeth between the two flocks.

Some aspects of thecal and granulosa cell function during follicular development in the bovine ovary.

The patterns of ovarian follicular development and the steroidogenic properties of individual follicles (greater than or equal to 2 mm diam.) were assessed in Angus cows from Day - 5 until Day + 1 of the oestrous cycle (oestrus = Day 0). Individual follicles were judged to be healthy or atretic using a new classification system incorporating assessments of thecal vascularity and colour, the number of granulosa cells, the presence or absence of debris in follicular fluid and the status of the oocyte. The results suggest that the theca interna of small antral follicles (less than 5 mm diam.) responds to LH and synthesizes androstenedione before the granulosa cells develop an appreciable ability to metabolize androgen to oestrogen. Regardless of follicle size, the output of thecal androstenedione per unit mass of tissue remained unchanged in healthy but not in atretic follicles. On a per cell basis, aromatase activity increased in granulosa cells from healthy but not from atretic follicles with increasing follicle size. Peak levels of aromatizing activity were consistently observed in dominant oestrogen-enriched follicles on Day 0 although similar activity was also observed in some healthy follicles (greater than or equal to 8 mm diam.) on other days of the cycle. Early atresia in bovine follicles was characterized by an absence or lowering of aromatase activity in granulosa cells which always preceded any reduction in the thecal steroidogenic response to LH. It was estimated that between 20 and 60 antral follicles (greater than or equal to 2 mm diam.) per cow may respond to LH by synthesizing androgen whereas only 1-3 follicles (greater than 5 mm diam.) have granulosa cells capable of metabolizing androstenedione or testosterone to oestradiol.

The pathogenesis of excessive wear in the deciduous teeth of sheep.

The teeth and various supporting and ancillary tissues were examined post mortem from two flocks of sheep, one of which had previously shown high deciduous incisor wear. Sheep were killed in groups selected on clinical signs at equivalent ages from birth to 116 weeks of age. At the end of this time the lengths of clinical crowns of the remaining deciduous incisor teeth in the high wear flock were not measurable, whereas the other flock had up to 5.5 mm in visible crown. Using the cingulum as a marker, measurements were made from extracted teeth of the attrition of the crown and size of the root. The tooth root did not grow substantially longer after the 13th week of the sheep;s life. Wear facets were just visible in some sheep at six weeks. More rapid wear occurred three times in the high wear flock. The first of these was relatively small and occurred between the 13th and 27th week of age in I1 and I2. The second was larger and occurred between the 38th and 57th week of age in the winter/spring period. A third period of wear, which was not measured from the cingulum, because this structure had by then been worn away, was seen in the remaining deciduous teeth between the ages of 91 and 110 weeks of age, i.e., during the second winter and spring. In the low wear flock the first of these periods of wear did not occur, the second was of shorter duration and smaller size, and the third equivocal. Eruption of the teeth after the 6th week was principally passive. There was no difference in the degree of wear of the molar teeth from the two flocks. Histological examination showed that the wear was compensated for by the deposition of reparative dentine in the pulp cavity. This usually gave the appearance of an ordered process. However, when wear was excessively rapid, a different, less orderly form of reparative dentine was laid down. In a few cases the reparative process was not fast enough, and a connection developed between the mouth and the pulp causing pulpitis. There was no histological evidence that the supportive tissues of the teeth, or the upper dental pad, contributed to the excessive wear.

Histological, macroscopic and microhardness observations of fluoride-induced changes in the enamel organ and enamel of sheep incisor teeth.

When aged 8.5 months, 10 sheep born in the same week were given 4 mg fluoride (F)/kg body weight orally for 26 days. Three sheep received no F. Sheep were killed at the end of the treatment period and later at selected stages of tooth development. The macroscopic changes in the enamel of one incisor were related to the cellular changes in the enamel organ of the contralateral tooth. A break in enamel continuity, hypoplasia, was seen on the labial enamel of 9 of the 10 F-treated sheep. Pitting of the enamel was associated with shortening of some ameloblasts and aggregations of cysts affecting cells late in their secretory phase in the first-killed sheep. In sheep killed later, these changes were associated with cells which had progressed into their maturation phase. A more extensive absence of enamel with ledge formation cervically, seen in one sheep, was associated with displacement or death of almost all the cells in their secretory phase during F treatment and consequent retention of the organic matrix. The hypoplastic lesions resulted from secretory-cell reaction during the period of F dosing. Diffuse patchy opacities, characterized by an irregular hypomineralized surface zone, were only apparent in the enamel of the later-killed sheep and were associated in one sheep with abnormal ameloblast regression in the contralateral tooth. These defects possibly resulted from the long-continued release of F stored in the bones during the period of F dosing.

Effects of long-term treatment with LH on induction of cyclic ovarian activity in seasonally anoestrous ewes.

The aim of the present study was to establish whether cyclic ovarian activity could be induced and then maintained in anoestrous Romney ewes by the long-term administration of regular intravenous pulses of LH (10 micrograms ovine LH i.v. once every 1 or 2 h for 29-91 days). The LH pulse regimen was designed to generate plasma profiles of LH that were comparable to those experienced during the luteal and follicular phases of the oestrous cycle. The results showed that the LH treatments were capable of inducing cyclic ovarian activity, as assessed from the concentrations of progesterone in plasma, but that the treatments were inadequate for sustaining cyclic activity beyond two consecutive progestational phases. After 35-56 days of treatment, the plasma concentrations of FSH declined significantly (P less than 0.05) relative to those in the untreated animals. These data suggest that FSH supplementation as well as LH might be required for the long-term maintenance of cyclic ovarian activity in seasonally anoestrous ewes.

Changes in gonadotrophin secretion and ovarian antral follicular activity in seasonally breeding sheep throughout the year.

Overall, significantly more antral follicles greater than or equal to 1 mm diameter were present in Romney ewes during anoestrus than in the breeding season (anoestrus, 35 +/- 3 (mean +/- s.e.m.) follicles per ewe, 23 sheep; Day 9-10 of oestrous cycle, 24 +/- 1 follicles per ewe, 22 sheep; P less than 0.01), although the mean numbers of preovulatory-sized follicles (greater than or equal to 5 mm diam.) were similar (anoestrus, 1.3 +/- 0.2 per ewe; oestrous cycle, 1.0 +/- 0.1 per ewe). The ability of ovarian follicles to synthesize oestradiol did not differ between anoestrus and the breeding season as assessed from the levels of extant aromatase enzyme activity in granulosa cells and steroid concentrations in follicular fluid. Although the mean plasma concentration of LH did not differ between anoestrus and the luteal phase of the breeding season, the pattern of LH secretion differed markedly; on Day 9-10 of the oestrous cycle there were significantly more (P less than 0.001) high-amplitude LH peaks (i.e. greater than or equal to 1 ng/ml) in plasma and significantly fewer (P less than 0.001) low amplitude peaks (less than 1 ng/ml) than in anoestrous ewes. Moreover, the mean concentrations of FSH and prolactin were significantly lower during the luteal phase of the cycle than during anoestrus (FSH, P less than 0.05, prolactin, P less than 0.001). It is concluded that, in Romney ewes, the levels of antral follicular activity change throughout the year in synchrony with the circannual patterns of prolactin and day-length. Also, these data support the notion that anovulation during seasonal anoestrus is due to a reduced frequency of high-amplitude LH discharges from the pituitary gland.

The prevalence and microbiology of pneumonia in a flock of lambs.

A flock of New Zealand Romney lambs on a property in Hawkes Bay was examined from August 1978 to June 1979. In November, the lambs in the flock were allocated to groups as follows: 600 lambs from which groups were selected for slaughter by commercial criteria each month (selected groups); 700 lambs which were subdivided into 8 groups (random groups), one of which was sent for slaughter each month; and 25 lambs to monitor serological responses to viral infections (surveillance group). At slaughter, lesions in the anterior lobes of the lungs were classified into small, large or mottled lesions while those in the posterior regions of the diaphragmatic lobes comprised a separate group. The prevalence of the pneumonic and pleural lesions was recorded in all groups of lambs. Pneumonic lesions in lambs from the random groups were further classified after microscopic examination. The prevalence of infections with parainfluenza virus type 3, adenoviruses, Mycoplasma spp., and Pasteurella haemolytica was also recorded in the random groups. Average carcase weights of all groups of lambs were also recorded. The prevalence of large lesions in the anterior lobes, pleural lesions and devaluation of carcases due to pleural lesions was significantly lower in the selected than in the random groups. In the random groups, the prevalence of small lesions in the anterior lobes of the lungs was high and was not always associated with infections with the viruses, P. haemolytica or Mycoplasma spp. Microscopically, these lesions could he divided into four types as previously described. The prevalence of large lesions in the anterior lobes was low in November but increased to peak levels in February and March and then declined to intermediate levels. The prevalence of large lesions was associated with that of infections with the micro-organisms and significantly more large lesions contained P. haemolytica and Mycoplasma spp. than did small lesions or normal lungs. The prevalence of large lesions also appeared to be inversely related to the average carcase weights of the lambs. Most large lesions were similar on microscopy to those of "enzootic" or "atypical" pneumonia but were divided into four types on the basis of exudative and proliferative features. The prevalence of mottled lesions in the anterior lobes was low throughout the period of observation but peaks were observed in January and May. The peaks were associated with adenovirus infection and elevated prevalence of infection with P. haemolytica and Mycoplasma spp. Significantly more mottled lesions contained P. haemolytica than did small lesions or normal lungs. Microscopically, most mottled lesions were similar to exudative large lesions. Lesions in the posterior diaphragmatic lobes were most numerous in November and from April to June. Their prevalence appeared to be related to anthelminthic treatment and environmental conditions. Microscopically, these lesions were typical of those following infection with Dictyocaulus filaria.

The production of developmental defects of enamel in the incisor teeth of penned sheep resulting from induced parasitism.

Thirty-four sheep were infected with nematode parasites (Trichostrongylus spp. and Ostertagia circumcincta) during central incisor formation. The tooth enamel was examined for defects of mineralization at eruption and compared with that of 14 control sheep. The type and number of parasites given, the duration of the infection and the age of the sheep were varied. Sheep receiving high doses of parasites e.g. 200,000 T. vitrinus + 20,000 O. circumcincta showed severe systemic effects (debilitation, scouring and weight loss) which were of short duration (7-10 days) as the infections were terminated by anthelmintic treatment. Hypoplastic enamel (a break in continuity) was induced in all 5 sheep infected at 8 1/4-9 months of age and demarcated opacities (an alteration in the translucency) in 3 out of 4 sheep infected at 10 months of age. Sheep given 150,000 or 40,000 T. colubriformis showed less obvious systemic effects although the weight loss continued in some sheep for up to 54 days. Although opacities of the enamel in the form of parallel lines, sometimes with localized increase in width or with a diffuse opacity superimposed, were seen by microscopic examination (X 10) of teeth in this group, many of the teeth from the control sheep showed similar lesions. Only the hypoplastic lesions can be attributed with confidence to the experimental manipulations.

Oat cells in the pathology of ovine pneumonia-pleurisy.

Light microscope observations on oat cells in the ovine pneumonia-pleurisy complex are presented. This study is based on the experimental production of the disease by viruses and Pasteurella haemolytica. Oat cells appeared only in necrotic lesions associated with large numbers of P. haemolytica in the pneumonic lung. It is suggested that oat cells originate from blood monocytes, which transform into the oat shape when developing in the necrotic, hypoxic environment created by P. haemolytica. They were not, however, observed to be phagocytic. Oat cells are characteristic of pneumonic pasteurellosis but are not pathognomonic because they can also be found in extrapulmonary locations and in other pathological conditions of the lungs.

Induction of cyclic ovarian activity in seasonally anoestrous ewes with exogenous GnRH.

Anoestrous ewes (N = 3) were treated with a 500 ng GnRH pulse administered via a jugular cannula every 2 h for 40 to 80 days. Plasma concentrations and therefore presumed ovarian activity changed cyclically with each progestational cycle (n = 10) lasting 14.0--18.5 days. It is concluded that, by increasing the frequency of GnRH secretory episodes from an apparent endogenous level of one episode per 3.6 h to at least one every 2.0 h, cyclic ovarian activity can be restored to seasonally anoestrous sheep.

Accumulation of luteinizing hormone, oestradiol and androstenedione by sheep ovarian follicles in vivo.

The temporal relationship between the levels of LH in peripheral plasma and in follicular fluid of ovarian follicles in anaesthetized sheep were investigated for a 10-h period after a single i.m. injection of LH releasing hormone (LH-RH; 100 microgram). The ovarian secretion rates of oestradiol and androstenedione and the levels of these steroids accumulating in different sized follicles at varying time-intervals after the LH-RH injection were also compared. The data show that the rates at which pituitary LH enters and leaves the intrafollicular fluid-filled spaces are substantially slower than those of peripheral blood. Two hours after LH-RH injection the levels of LH in plasma had increased from 1 to 200 ng/ml, whereas in the follicle the levels remained at approximately 2ng/ml. Ten hours after the LH-RH injection, the levels of LH in plasma had returned to basal values (approximately 1.4 ng/ml) but in both small and large follicles the levels of LH (approximately 20 ng/ml) were comparable to those present in similar sized follicles 4h earlier. The data also indicate that more than 90% of the oestradiol produced by a large antral follicle (greater than or equal to 5 mm diameter) probably enters the bloodstream without first accumulating within the follicular antrum. Finally it is concluded that the clearance of the small amount of oestradiol which does accumulate in the follicular antrum is negligible compared with the clearance of this hormone from peripheral plasma.

Evidence that changes in luteinizing hormone secretion regulate the growth of the preovulatory follicle in the ewe.

The aim of the present study was to gain evidence that the level of LH secretion preceding the preovulatory LH surge is an important determinant of follicular maturation and corpus luteum function in the ewe. In addition it was hoped to establish whether the pattern of LH delivery to the ovary (pulsatile v. constant) is a critical factor in the maturation of a preovulatory follicle. To accomplish this, progesterone-primed anoestrous ewes were repeatedly injected i.v. with LH or luteinizing hormone-releasing hormone (LH-RH), or given an i.v. infusion of LH, over a 72 h period. These animals, together with the appropriate controls, were exposed to a sexually active ram so that oestrous activity could be recorded. All ewes were subjected to intensive blood sampling regimes so that the plasma levels of LH and progesterone could be determined and compared to those which occurred in the same breed of sheep during the oestrous cycle. Collectively the data suggest that the plasma levels of LH preceding the preovulatory LH surge are an important determinant of follicular maturation as judged by subsequent corpus luteum function. Moreover, they show that follicular maturation can be achieved with widely differing patterns of LH delivery to the ovary during the preovulatory period and that a strict pulsatile delivery of LH may not be an absolute requirement.