The embryonic development of the bovine stomach revisited.

The adult anatomy and physiology of the bovine (Bos taurus) stomach have been investigated extensively. Despite the many studies, however, the early development of the stomach has not yet been fully elucidated. The goal of the present study, therefore, was to review the available literature, to visualize the embryonic and early foetal development of the bovine stomach and to shed light on unresolved issues. The stomachs of fifteen bovine embryos and eleven foetuses from 26 to 80 days of gestation were photographed both in situ and after exenteration and critical point drying. A series of photographs was obtained that yielded a contiguous and comprehensive view of all the developmental changes that occurred until the virtually final configuration of the stomach was attained. In addition, the serosal surface was studied by electron microscopy, thus revealing subtle regional differences in the lining of the peritoneal cavity. Our observations corroborate the contention that all the compartments evolve from the fusiform primordium and that no outgrowth at the level of the oesophagus occurs. The greater curvature as well as the attachment line of the dorsal mesogastrium shift to the left, which is similar to the process in monogastrians. The rumen and reticulum develop from separate protrusions, and further compartmentalization results from constrictions and bulges and not from folding. Between 55 and 60 days of gestation, the entire bovine stomach except for the abomasum eventually relocates to its final position. In summary, previously debated key issues were addressed and integrated with current findings.

Computed tomography study of the fetal development of the dairy cow stomach complex.

In the fetal development of animals, critical physiological and anatomical events influence the long-term health and performance of the offspring. To identify the critical growth phases of the fetal bovine stomach, we used computed tomography imaging on 30 German Holstein fetuses to examine the fetal bovine stomach in situ. Computed tomography allows the study of diverse parameters such as the volume of the stomach chambers in situ without the need for sophisticated filling preparation techniques. The absolute volume, relative volume, and monthly volume increase of each stomach chamber were determined. Computed tomography was a reliable method for in situ examination of the fetal bovine stomach complex from the third month of gestation onward. It was able to detect an abnormal position of the abomasum in 2 fetuses. The crown-rump length of the fetuses studied ranged from 9.5 to 89 cm (from 2.2 to 8.3 mo of gestation). Over this timeline, the changes in the relative volumes of the ruminoreticulum and abomasum were inversely related. Until mo 5 of gestation, the relative volume of the ruminoreticulum increased steadily, whereas that of the abomasum decreased. Thereafter, the relative volume of the ruminoreticulum became gradually smaller, and that of the abomasum became larger; by mo 8, the abomasum was larger than the ruminoreticulum. All stomach chambers had large increases in volume over the gestation period and we observed differences in development patterns and volume changes of the individual stomach chambers over this period. The largest monthly volume increase of the stomach complex was between mo 4 and 5 of gestation. In this period, the volume of the ruminoreticulum increased 43.8 times, that of the omasum 38.9 times, and that of the abomasum 30.03 times. Between mo 5 and 6 of gestation, the abomasum had another growth spurt, with a monthly volume increase of 10.4 times. These 2 time points in the gestation period may be critical phases of fetal development that should be considered in the management of pregnant cattle.

Amplicon sequencing of bacterial microbiota in abortion material from cattle.

Abortions in cattle have a significant economic impact on animal husbandry and require prompt diagnosis for surveillance of epizootic infectious agents. Since most abortions are not epizootic but sporadic with often undetected etiologies, this study examined the bacterial community present in the placenta (PL, n = 32) and fetal abomasal content (AC, n = 49) in 64 cases of bovine abortion by next generation sequencing (NGS) of the 16S rRNA gene. The PL and AC from three fetuses of dams that died from non-infectious reasons were included as controls. All samples were analyzed by bacterial culture, and 17 were examined by histopathology. We observed 922 OTUs overall and 267 taxa at the genus level. No detectable bacterial DNA was present in the control samples. The microbial profiles of the PL and AC differed significantly, both in their composition (PERMANOVA), species richness and Chao-1 (Mann-Whitney test). In both organs, Pseudomonas was the most abundant genus. The combination of NGS and culture identified opportunistic pathogens of interest in placentas with lesions, such as Vibrio metschnikovii, Streptococcus uberis, Lactococcus lactis and Escherichia coli. In placentas with lesions where culturing was unsuccessful, Pseudomonas and unidentified Aeromonadaceae were identified by NGS displaying high number of reads. Three cases with multiple possible etiologies and placentas presenting lesions were detected by NGS. Amplicon sequencing has the potential to uncover unknown etiological agents. These new insights on cattle abortion extend our focus to previously understudied opportunistic abortive bacteria.

Comparative analysis of the merino sheep and Iberian red deer abomasum during prenatal development.

The aim of this study is to describe differences in the ontogenesis of the abomasum in sheep (domestic ruminant) and deer (wild ruminant). Histomorphometric and immunohistochemical analysis were carried out on 50 embryos and fetuses of the sheep and 50 red deer from the first prenatal stages until birth. To compare similar periods of gestation in both species, we calculate the percentages of gestation. The appearance of the abomasum was earlier in the red deer (22% gestation) than in the sheep (25% gestation). Throughout development the epithelium happened sequentially, being of the types pseudostratified to simple cylindrical. This important modification was earlier in the red deer than the sheep. At 46% gestation in red deer and 50% in sheep, gastric pits were observed on the surface of abomasal folds. Our studies suggest a close link between the initial formation of these pseudoglandular structures and the clear separation of lamina propria and submucosa separated by de muscularis mucosae. At 54% gestation in red deer and at 60% in sheep, in the bottom of these pits the first outlines of glands were distinguishable. Finally, the presence of neuroendocrine and glial cells were detected in deer at earlier stages than in sheep.

Histomorphometric study of the goat stomach during prenatal development.

The aim of this study was to perform a morphometric analysis of the different structural tissue layers of the goat stomach to study their prenatal growth from mathematical models fitted to these morphometric data. A total of 90 embryos and fetuses were used, from the early stages of prenatal life until birth. The growth rate of the gastric wall was slower than that of body length; rumen was the stomach compartment displaying slowest growth. In the three non-glandular compartments, the epithelial layer grew faster than the gastric wall itself, while the growth rate of the abomasal epithelium declined in the early stages of development. A decline in growth rate was also observed for the lamina propria and submucosa in rumen and reticulum from the early embryonic stages, whereas in omasum and abomasum these layers continued to grow as gestation progressed. The tunica muscularis displayed consistent growth in all compartments, growing faster than the gastric wall. Serosa thickness increased as gestation progressed, displaying a decline in growth-rate only in the omasum. In conclusion, the dynamics of gastric wall growth were governed by the growth rate of each of the component tissue layers.

Histomorphometric and immunohistochemical study of the goat abomasum during prenatal development.

This study sought to chart the morphological changes taking place in the goat abomasum during prenatal development, using histomorphometric and immunohistochemical techniques. A total of 140 goat embryos and fetuses, from the first stages of prenatal life until birth. Differentiation of the abomasum as a separate compartment of the primitive gastric tube was observed at 35 days of prenatal life (CRL 3 cm, 23% gestation). Primitive abomasal folds were first observed at 38 days (CRL 4.3 cm, 25% gestation). The muscularis mucosae was visible by 64 days (CRL 13.5 cm, 43% gestation). Transformation of pseudostratified epithelium to simple cylindrical epithelium was also observed at this stage. Differentiation of gastric pits and glands first became apparent at 75 days (CRL 17.5 cm, 50% gestation) and 84 days (CRL 20 cm, 55% gestation), respectively. Neuroendocrine cells were detected by synaptophysin (SYP) at 64 days (CRL 13.5 cm, 43% gestation), while glial cell markers (glial fibrillary acidic protein - GFAP, and vimentin-VIM) were observed at 64 days (CRL 13.5 cm, 43% gestation) and 38 days (CRL 4.3 cm, 25% gestation), respectively. Neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) were detected at 75 days (CRL 17.5 cm, 50% gestation). Gastrin-immunoreactive cells first appeared in the abomasum at 76 days (CRL 18 cm, 50% gestation). In conclusion, prenatal development of the abomasum appears to take place somewhat earlier in goats than in sheep or cattle, but at a similar rate to that reported in wild ruminants such as deer.

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.

Bovine neosporosis: immunoblot improves foetal serology.

The purpose of the study is the comparative evaluation of the immunofluorescent antibody test (IFAT) and an immunoblot (IB) test for the examination of foetal fluids for specific antibodies against Neospora caninum. Peritoneal and pleural fluids as well as abomasal contents were analysed. The results of the serological examinations were compared to those obtained by histological, immunohistochemical, and PCR analysis of foetal tissues as well as to the results of maternal serological examinations. Fluids were used undiluted in the IB and reactions against six immunodominant antigens were recorded. When the recognition of at least two immunodominant antigens was regarded as positive, the agreement of the IB with other diagnostic methods was good to moderate as characterised by kappa-values of 0.76 (histology/immunohistochemistry), 0.69 (maternal serology) and 0.54 (PCR on foetal tissues). The IB results agreed better with the results of the other diagnostic methods than those of the IFAT. The higher relative sensitivity of the IB was regarded as the main reason for the better agreement. However, also the specificity of the IB was superior to that of the IFAT in relation to histology/immunohistochemistry, maternal serology and PCR.

Proliferation in the gastric epithelium of bovine abomasum during foetal development as revealed by Ki-67 immunocytochemistry.

For the first time, proliferative cells in the abomasal mucosa of 30 bovine foetuses with a crown-rump length of 34 to 1000 mm (approximately 45-250 days of gestation) were detected using the antibody MIB-1 against the (human) nuclear-associated protein Ki-67. At the beginning of mucosal development, the epithelium is stratified and MIB-1 positive cells are scattered all over the epithelium. With the formation of gastric pit/gland invaginations, the proliferative cells are mostly confined to the basal half or third of the gastric pit/gland unit, both in the regio glandulae propriae and in the regio glandulae pyloricae. From the 27th week in the regio glandulae propriae and the 30th week in the pyloric gland region, respectively, the proliferative cells move upwards and are situated in the bottom of the pit and the upper part of the gland, whereas the base of the gland is free of labelling.

An immunohistochemical study on the development of progastricsin-immunoreactive cells in the bovine abomasal mucosa.

The development of progastricsin-immunoreactive cells in the abomasal mucosa of cattle fetuses was studied by immunohistochemistry. Progastricsin-immunoreactive cells were detected first in the fundic and pyloric regions of 52 cm in crown-rump length (CRL) fetuses (about 180 days of gestation). The frequency of occurrence of progastricsin-immunoreactive cells and the intensity of their immunoreactivity increased with the progress of gestation, but most of these immunoreactivities were restricted to the basal portion of the fundic and pyloric glands. After birth, in the fundic mucosa, progastricsin immunoreactivities were found not only in the chief cells but also in the surface mucous cells of the gastric pits. In the cow, the immunoreactivity of the surface mucous cells was even stronger than that of the chief cells. In the pyloric mucosa, progastricsin immunoreactivities were found in the gastric pits and in the basal portion of the pyloric glands, but they were organized in small groups and showed a patchy distribution in the basal portions.

Maturational effects of cortisol on the exocrine abomasum and pancreas in fetal sheep.

The role of cortisol in the prenatal development of digestive enzymes in the abomasum (prochymosin and pepsinogen) and pancreas (amylase, trypsin, chymotrypsin) has been investigated in the fetal lamb during late gestation. The abomasum and pancreas were collected from 22 unoperated control fetuses (99-145 days gestation; term, 145 +/- 2 days), from seven pairs of twins infused with either saline or cortisol for five days preceding delivery at 127-133 days, and from four 139-143-day-old fetuses adrenalectomized at 120-123 days. Developmental increases (2-8-fold) occurred in protease concentrations in the fetal abomasum and in amylase and chymotrypsin contents in the fetal pancreas. These increases paralleled the normal prepartum rise in fetal plasma cortisol. In addition, the enzyme values were significantly higher in cortisol-infused than in saline-infused fetuses (with the exception of pancreatic amylase) and were significantly lower in adrenalectomized fetuses than in control fetuses at term. The pH of abomasal fluid remained neutral (pH 6.8-8.0) during late gestation and was not affected by cortisol treatment or adrenalectomy. The results suggest that cortisol stimulates the development of the exocrine abomasum and pancreas in fetal sheep and may, thereby, increase the digestive capacity in neonatal lambs. Compared with the pig, another long-gestation species, the sheep has an early development of gastric pepsinogen but a late development of gastric acidity and pancreatic protease activities.

Differentiation of oxyntic cells during early ontogenesis in the bovine.

The origin and the differentiation of oxyntic cells in fetal bovine abomasum were investigated using transmission electron and light microscopic methods. In the oxyntic gland region oxyntic cell precursors and immature oxyntic cells appear as early as at the end of the first trimester of gestation--much earlier than described in any other mammalian animal species. Immature oxyntic cells are characterized by long apical microvilli, by their triangular-shaped light cytoplasm rich in large and numerous mitochondria, by the existence of vesicular profiles and by the incipient invagination of the apical plasma membrane forming a primitive intracellular canaliculus expanding into central areas of the cell. The oxyntic cell represents the first exocrine cell type developing from secretory granule-containing cells in the base of the primitive gastric glands.

Histomorphometric analysis of the abomasum of the sheep during development.

Histomorphometric analyses were carried out on 64 embryos and fetuses and on 20 sheep (early postnatal to adult age). Histodifferentiation of the abomasum took place at 33 days of fetal life, with the appearance of abomasal villi at 53 days. By 64 fetal days, the epithelium had changed from pseudostratified to simple mucous cylindrical. Acidic glycoproteins appeared at 46 fetal days. Neutral glycoproteins did not appear until later stages of development, near birth. We believe that the configuration of a simple epithelium with acidic secretion is enhanced at birth by the secretion of neutral glycoproteins which act as a buffer against acidic substances, and particularly against the abomasal acidity during lactation. Growth curves and formulae were set out for each tissue layer.

Production of prochymosin, pepsinogen and progastricsin, and their cellular and intracellular localization in bovine abomasal mucosa.

A brief overview of research is presented on the production, cellular and intracellular localization of prochymosin, pepsinogen and progastricsin in bovine abomasal mucosa from fetus to adult. Prochymosin is produced early during gestation (10th week) and is significantly related to milk-feeding. Pepsinogen and progastricsin start to be produced later during gestation (20th week) and are produced in low amounts as long as the calf is fed milk. With age, pepsinogen becomes the dominating zymogen in the abomasal mucosa. Most of the cell types in the fundic gland have the ability to produce all three zymogens and are also found in the same individual secretory granules of these cells.

[The development of the bovine stomach: morphologic and morphometric analysis. II. Observations of the morphogenesis associated with the omasum and abomasum]. / El desarrollo del estómago del vacuno: análisis desde el prisma morfológico y morfométrico. Parte II: Observaciones morfogénicas asociadas al bloque omaso-abomásico.

This study represents a continuation of part 1 of a study of the development of the bovine stomach. Material and methods are more fully described in part i of the study. In this part 2 of the study further developmental stages and morphometric analysis is presented.

[The development of the bovine stomach: morphologic and morphometric analysis. III. Spatial and morphometric reconstruction]. / El desarrollo del estómago del vacuno: análisis desde el prisma morfológico y morfométrico. Parte III: Reconstrucción espacial y morfometría.

This work represents a summary of the study presented in parts I and II, previously published in this journal. The data collected in those studies is presented in three dimensional reconstructions of the stomach of the developing calf.

Histogenetic evolution of bovine gastric compartments during the prenatal period.

In order to carry out this study, 62 embryos and fetuses of Friesian cows were used, and every stomach was dissected and subjected to the usual laboratory procedures. After obtaining each preparation all the histological and metrical changes that took place in the wall of the viscus during its development are described. The compartments are histologically differentiated into three stages: in the first one there is no compartment differentiation: in the second one, the rumino-reticulum, omasum and abomasum have differentiated; and in the third one every compartment is differentiated. In group 16, (78 days of gestation), a decrease in thickness of the muscular layer of all compartments, parallel to the histological differentiation has been observed.

Histological development of bovine abomasum.

The histological development of the bovine abomasum during fetal and neonatal periods was studied. The abomasum in a fetus of 2.3 cm in length (estimated to be 1 month old) was a separated compartment situated to be caudo-ventral to the primordium on the median plane. On the later stages, the primitive stomach became distinctly separated into rumen, reticulum, omasum and abomasum. The epithelium of abomasum was pseudostratified histologically, consisting of cells low differentiation. The stomach in a fetus of 13-14 cm in length (estimated to be 3 months old) displayed the morphological feature with nearly same proportion as an adult stomach. In this stage, abomasal epithelium had a shape of simple column and also formed gastric pits. Pyloric gland cells could be recognized earliest of all the gastric exocrine cells in fetuses of 16-18 cm in length (estimated to be 3-4 months old). Mucous neck cells could be found in fetuses 43-45 cm in length (estimated to be 5-6 months old). Pyloric gland cell and mucous neck cell contained neutral and sialo- or sulfo mucosubstances in neonates. Chief cells, could be noticed in fetuses 58-65 cm in length (estimated to be 6-7 months old), and were devoid of demonstrable mucosubstance as well as parietal cells. Main abomasal gland cells began to develop to increase rapidly in number in the latter half period of gestation. All the types of gastric cells became to be present and mature in form at birth.