Cm Crown-rump Length Research Articles

Transplacental transmission of Neospora caninum in moose (Alces alces).

A 5-years-old moose (Alces alces) cow kept in a zoo in the German Federal State of Brandenburg aborted a female foetus of 44cm crown rump length (CRL). Pathohistological analysis revealed several Neospora (N.) caninum infected cells and cysts, as well as multifocal gliosis, necrosis, haemorrhages, dystrophic mineralisation and haemosiderosis in the brain, predominantly in cerebrum and brainstem. In addition, mild lymphocytic meningitis was present. Together with the fresh foetus, a mummified foetus of 16cm CRL was expelled. Neither focal necrosis, nor inflammation was detected in the brain of the mummified foetus. By two polymerase chain reactions (PCR) targeting the pNc5 gene of N. caninum (i.e. an end point PCR and a real-time PCR), by two serological methods (immunofluorescence test and immunoblot), by histological and immunohistochemical analyses, transplacental N. caninum infection was confirmed in the fresh foetus and interpreted as possible cause of abortion. Infection with other agents causing abortion including Bovine Herpesvirus 1 (BHV1), Bluetongue Virus (BTV), Bovine Virus Diarrhoea Virus (BVDV), Brucella spp., Chlamydia spp., Coxiella burnetii and Toxoplasma gondii were excluded. Our findings show that control measures may be necessary to protect captive moose against accidental N. caninum infection. Further studies are needed to explore the importance of neosporosis in wild and captive moose.

Prenatal development of the bovine epididymis: Light microscopical, glycohistochemical and immunohistochemical studies

Prenatal development of the epididymis was studied in bovine fetuses ranging from 10 to 90cm crown-rump length (CRL) (75–285 pcd). The studies aimed to apply both glycohistochemistry and immunohistochemistry for the detection of the differentiation of the developing prenatal epididymis. Both conventional histological and histochemical techniques were applied on paraffin sections of the epididymis from different fetal stages. Establishment of the urogenital junction between the extra-testicular rete testis and the mesonephric duct, via the growing efferent ductules (ductuli efferentes) was first evident in fetuses with 10cm CRL. At the fetal age of 110 pcd (24cm CRL), the mesonephric duct began to lengthen and coil forming three distinct regions (caput, corpus and cauda). In addition to the macroscopical modifications in the extra-testicular excurrent duct system, histological differentiation involved both the tubular epithelial and the peritubular mesenchymal cells. The epithelium lining the efferent ductules was differentiated into ciliated and non-ciliated columnar cells. The simple epithelium of the epididymal duct increased in height and developed stereocilia on the apical surface. Additionally, some basal cells first appeared at 185 pcd (56cm CRL), within the epithelium lining the cauda only. Lectin histochemistry (WGA, PNA, GSA-I) showed early immunostaining in epithelium of the efferent ductules and in peritubular mesenchymal structures. Immunoreactivity for different proteins (S-100, fibroblast growth factor-1 and factor-2, angiotensin converting enzyme, laminin, alpha-smooth muscle actin) was evident, both in the epithelial and in the peritubular mesenchymal cells as early as at 75 pcd. On the basis of our histochemical observations, we conclude that both glycohistochemistry and immunohistochemistry are useful tools to demonstrate that the differentiation in the peritubular structures and efferent ductular epithelium begins earlier than other components.

Quantification, Morphology and Ultrastructure of Preantral Follicles of Buffalo (Bubalus bubalis) Foetuses

The objective of this study was to determine the number, morphology and ultrastructure of preantral ovarian follicles of buffalo (Bubalus bubalis) foetuses at different ages. Quantification revealed number of primordial, primary and secondary follicles of 48,857 ± 17,506, 26,000 ± 20,452, 18,428 ± 10,875 and 18,375 ± 19,690, 225 ± 349, 326 ± 288 at 12-34 cm and 35-60 cm crown rump length (CRL), respectively. Follicular diameter values were 28.9 (± 3.4), 34.7 (± 5.9) and 59.4 (± 12.6) μm; oocyte diameters were 21.7 (± 2.8), 24.3 (± 3.4) and 33.0 (± 7.7) μm, and the numbers of follicular cells in the follicle equatorial section were 7.1 (± 1.4), 12.0 (± 2.4) and 13.8 (± 2.4) for primordial, primary and secondary follicles, respectively. The primordial follicle consisted of an oocyte surrounded by a layer of flattened follicular cells with a normally eccentric oocyte nucleus. Dispersed Golgi complex, smooth endoplasmic reticulum, rounded mitochondria and several lipid vesicles were observed in the cytoplasm and cell junctions between the follicle cell membranes and the oocyte. This work describes the number, morphometry and ultrastructure of preantral follicles of buffalo foetuses, concluding that folliculogenesis is established between 8 and 34 cm CRL and that follicle number varies individually and according to age and that further studies are needed in this species.

Morphometric and immunohistochemical study of the abomasum of red deer during prenatal development

The red deer is well suited to scientific study, given its economic importance as an animal to be hunted, and because it has a rich genetic heritage. However, there has been little research into the prenatal development of the stomach of ruminants in general, and none for the red deer. For this reason, we undertook histological evaluation of the ontogenesis of the abomasum in red deer. Histomorphometric and immunohistochemical analyses were carried out on 50 embryos and fetuses from the initial stages of prenatal life until birth. The animals were divided for test purposes into five experimental groups: group I [1.4-3.6 cm crown-rump length (CRL); 30-60 days, 1-25% of gestation]; group II (4.5-7.2 cm CRL; 67-90 days, 25-35% of gestation); group III (8-19 cm CRL; 97-135 days, 35-50% of gestation); group IV (21-33 cm CRL; 142-191 days, 50-70% of gestation) group V (36-40 cm CRL; 205-235 days, 75-100% of gestation). In the organogenesis of the primitive gastric tube of red deer, differentiation of the abomasum took place at 67 days, forming a three-layered structure: the epithelial layer (pseudostratified), pluripotential blastemic tissue and serosa. The abomasal wall displayed the primitive folds of the abomasum and by 97 days abomasal peak areas were observed on the fold surface. At 135 days the abomasal surface showed a single mucous cylindrical epithelium, and gastric pits were observed in the spaces between abomasal areas. At the bottom of these pits the first outlines of glands could be observed. The histodifferentiation of the lamina propria-submucosa, tunica muscularis and serosa showed patterns similar to those described for the forestomach of red deer. The abomasum of red deer during prenatal life, especially from 67 days of gestation, was shown to be an active structure with full secretory capacity. Its histological development, its secretory capacity (as revealed by the presence of neutral mucopolysaccharides) and its neuroendocrine nature (as revealed by the presence of positive non-neuronal enolase cells and the neuropeptides vasoactive intestinal peptide and neuropeptide Y) were in line with the development of the rumen, reticulum and omasum. Gastrin-immunoreactive cells first appeared in the abomasum at 142 days, and the number of positive cells increased during development. As for the number of gastrin cells, plasma gastrin concentrations increased throughout prenatal life. However, its prenatal development was later than that of the abomasum in sheep, goat and cow.

Uterine Torsion in a West African Dwarf Ewe in Ibadan, Nigeria

Keywords: Uterine torsion, West African Dwarf Ewe, Ibadan A case of uterine torsion in a 21/2 year old pluriparous West Africa Dwarf (WAD) ewe raised semi intensively with adequate veterinary care before the death of the dam and the lamb is presented. The dam had been off feed for 3 days and was found straining a night before her death. The lamb was a male of 37cm crown rump length (CRL), weight, 3.0kg and showed other features consistent with a full term lamb. The post mortem findings and the predisposing factors are discussed. Trop. Vet. Vol. 20(4) 2002: 201-203

Entwicklung, Zytoarchitektonik und Feinstruktur des mesencephalen Trigeminuskerns bei Hauswiederkäuern *

The ontogenetic development and cell differentiation of the mesencephalic trigeminal nucleus (Ntm) is lightmicroscopically examined in 58 bovine embryos and fetuses ranging from 2.4 to 80 cm Crown-Rump-Length (CRL). The cytoarchitecture and fine structure in adult cattle, sheep, and goats are investigated with the aid of light- and electronmicroscopy. At 2.4 cm CRL, the proneurons of the Ntm are detectable for the first time within the ventricular zone of the alar plate, possessing one drop-like cytoplasmic protrusion, whereas at 5 cm CRL, two cell types with differing sizes appear. Up to a CRL of 11.5 cm, the nucleus shows advanced maturation processes and has reached his final position at the border of the mesencephalic central grey. From 26 cm CRL onward, three cell types, and at 34 cm CRL four cell types, are discernible based on their nissl-granule arrangement. The cytomorphological differentiation and the maturation of the cells proceeds until 56 cm CRL, at which point the topographical and cytological characteristics of the Ntm are comparable with those of adult animals. In adult cattle, sheep and goats the Ntm consists of large (40-60 microns) and scarce medium-sized (30-40 microns) neurons with round and oval shapes. Scarcer small (20-25 microns) round and medium-sized multipolar neurons occur. The Nissl bodies are scattered throughout the pericaryon of the large neurons in a dust-like pattern and in the medium-sized neurons in a grained form. Within the cytoplasmic streets, which are situated between the membranes of the rough ER, numerous neurofilaments and mitochondria are detectable. Large Golgi complexes are placed in a perinuclear position. The neurons are also characterized by some somatic spines, and by a moderate distribution of axosomatic synapses, in which axon-endings with flattened synaptic vesicles predominate.

Distributions of Extracellular Matrix and Carbonic Anhydrase-III during Bovine Palatine Ridge Development.

The present study describes histological alterations and immunohistochemical distributions of extracellular matrices (ECMs) and the carbonic anhydrase isozyme-III (CA-III) during the period of bovine palatine ridge formation. Morphogenesis of bovine palatine ridges was preceded by epidermal placodes and the mesenchymal condensation (MC). During the early stages of less than 44 cm crown rump length (CRL), fibronectin (FN) was distributed densely in the MC. Strong reactions against type I collagen (C-1) were detected outer to the FN positive site. In the stages of more than 44 cm CRL, FN and C-1 were distributed diffusely in subepithelial mesenchyme. Laminin (LN) and type IV collagen were distributed in the epithelial and endothelial basement membranes (BMs) in all of the stages examined, except in the stage of 7 cm CRL, where LN was not detected only in the BM just beneath the epidermal placode. CA-III was detected in basal epithelial cells except for palatine ridge rudiments in the stages of more than 21 cm CRL. It is suggested that the expressions of LN and CA-III might play a role in the spatial determination of rudiments of bovine fetal palatine ridges.

Distributions of Fibronectin, Laminin, Type I and IV Collagens and Carbonic Anhydrase Isozyme III during Bovine Ruminal Epithelial Development

The present study describes the immunohistochemical distributions of extracellular matrices and the carbonic anhydrase isozyme III (CA-3) in the bovine ruminal epithelium during fetal development. Fibronectin (FN), laminin and type I and IV collagens were distributed in the ruminal subepithelial mesenchymal regions with nonspecific regionality during the gestational periods. At stages after about 59 cm crown-rump length (CRL), FN of the epithelial basement membrane disappeared, and CA-3 appeared in the basal epithelial cells of the root of the ruminal papillae, suggesting that the functional differentiation of the ruminal epithelium might start at around 59 cm CRL in bovine fetuses.

Studies on the Ovarian Development in Zebu Cattle (Bos indicus)

In a study of 71 female foetuses, gonadal blastema was observed at 1.5 cm crown rump length (CRL). Oogonial cells entered the meiotic prophase at 3.5-6.0 cm CRL, which was arrested at the dictyotene stage to produce primary oocytes which formed primordial follicles. Primordial follicles were observed at 6-8 cm CRL. All germinal cells were at dictyotene by 20-24 cm CRL and follicles developed to primary and secondary stages. Folliculogenesis dominated further ovarian development and reached a peak between 32 and 35 cm CRL. In seven of the 12 foetuses measuring between 41 and 72 cm CRL, many follicles were atretic and some luteinized. The luteal bodies were composed of hypertrophied theca and granulosa cells with homogeneous and eosinophilic cytoplasm.

Correlation of placental erythrocyte morphology and gestational age.

A series of uncomplicated pregnancies presenting during the first trimester for elective termination was assigned a gestational age using accepted ultrasonographic nomograms. Following termination by suction curettage, the proportion of nucleated erythrocytes (NRBC) and anucleated erythrocytes (RBC) present in the placental circulation was estimated from hematoxylin and eosin-stained slides of the chorion frondosum using a 10 point scale. The score correlated tightly with sonographically assigned gestational age assessment between 15 and 45 cm crown-rump length (CRL) (48 and 83 days of gestation). These findings suggest the following: (1) the maturation of RBCs follows a course that is tightly correlated with gestational age; (2) an assessment of RBC morphology and, thus, an estimate of gestational age can be provided using routine histopathologic techniques and study of the placenta only; (3) this method may prove to be particularly useful in cases of unexplained spontaneous losses as a means of investigation when cessation of fetal development occurred.

The ontogeny of the thyrotropin-thyroid axis in early bovine embryos.

T4 and T3 formation and their response to TSH, cAMP, and prostaglandin E2 (PGE2) stimulation were studied by RIA in cultured bovine fetal thyroids from 130 embryos of 1.2-25.0 cm crown-rump length (CRL). T4 and T3 were found in all of the freshly isolated glands studied, and their concentrations increased significantly (P less than 0.05) during a 24-h incubation of glands from fetuses with a CRL greater than 8.0 cm. The release of T4 (nanograms per mg tissue), but not of T3, increased consistently with CRL (r = 0.64; P less than 0.05). The addition of TSH (0.5 mU) to the culture medium induced a 2- to 3-fold increase in the secretion of T4, but not of T3, by glands of fetuses with a CRL of 3.0-25.0 cm (P less than 0.05). Dibutyryl cAMP (10(-4) M) and PGE2 (10(-4) M) had comparable effects. A combination of TSH and theophylline (1 mM) or of TSH and dibutyryl cAMP significantly enhanced the T4 released by cultured tissue into the medium (P less than 0.05) over that induced by either agonist, but the combined effect was not fully additive. The total PGE2 in the tissue and medium was not changed by the addition of 0.5 mU TSH, and indomethacin had no effect on TSH-induced T4 secretion. The data show that thyroids of fetuses that have reached a CRL of 3.0 cm have the enzymatic capacity to produce both T4 and T3, and T4 is the dominant product formed. While exogenous PGE2 at high concentrations stimulates fetal T4 secretion, it does not mediate the actions of TSH and cAMP on the fetal thyroid.

Estradiol-17 beta biosynthesis by the early bovine fetal ovary during the active and refractory phases.

Estradiol-17i3 biosynthesis was studied by radioimmunoasssy during the “active” phase of ovaries of early bovine fetuses of 5-8 cm crown-rump length (CRL) (n = 40) and during the “refractory” phase of ovaries of fetuses of 10-20 cm CRL (n = 85). The mean value of estradiol17f3 secreted by the incubated ovaries of the early active phase was 1.5 ± 0.4 ng/ovary/24 h (mean ± SEM), and in the presence of bovine LH estradiol-1713 secretion was significantly enhanced (4.2 ± 0.6, P<0.OO1). In the presence or absence of LH, estradiol-17�3 was undetectable in the incubated ovaries of the refractory phase. Addition of testosterone to the incubation medium significantly enhanced estradiol-17(3 production (P<0.001) by ovaries of both the active and refractory phase. In contrast, neither progesterone, DHT, nor 17n-hydroxyprogesterone affected estradiol secretion. In the time-course studies of estradiol-17j3 production in response to testosterone, the incubated active phase ovaries from fetuses of 5-8 cm CRL responded to testosterone within 30 mm, whereas for the refractory phase ovaries a lag period of 10 h was needed. Addition of 8-Br-cAMP caused a significant stimulation after 5 h of both testosterone (P<O.O01) and estradiol (P<0.02) production by the incubated active ovaries, whereas when the inactive ovaries were used only progesterone secretion was significantly enhanced (P<0.0O1). The data suggest that the lack of ovarian aromatizable androgens may play a role in the change from the active to the refractory phase. That ovariesfrom bovine fetuses of 10-20 cm CRL become refractory suggests that the high estrogen level found at the time of the differentiation of the primordial gonad to ovary isof functional significance.

The early differentiation of the testis and interstitial cells in the fetal pig, and its duplication in organ culture.

AbstractThe unusual histogenesis of the fetal pig testis was examined as part of a study of its role in reproductive biology. Three types of interstitial cells were defined and their numbers estimated per unit area of testicular tissue in 6 μ‐serial sections. Statistical analysis of the percentage frequency of each cell type showed that the mature Leydig cells (C‐type) reached a peak at about 4.5 cm crown‐rump length (CRL) fetus and then fell to a low level, rising again sharply from 20 cm CRL until near birth. It was postulated that the first mature Leydig (C‐type) cells arose from mesenchymal (A‐type) cells which differentiated first into immature Leydig (B‐type) cells, and the process was repeated in the last part of fetal life.Mixtures of cock plasma, chicken embryo extract and balanced salt solution allowed maintenance and differentiation of whole fetal testes or testicular slices as organ cultures in Maximow slide assemblies for 6 to 14 days. Medium 199 with 10% bovine serum allowed maintenance and differentiation in some explants for at least six days. Indifferent gonads developed into recognizable testes or ovaries. Testes from 2.5, 3.0, and 3.5 cm CRL fetuses differentiated at a fairly normal rate, and some of their interstitial cells underwent the transformation postulated from the in vivo study. It was concluded that organ cultures from indifferent gonads and early testes provided living, functioning systems which could be used to test for androgenic activity with biological indicators.