Gross Anatomical Features of the Gastrointestinal Tract (GIT) of Blue‐and‐Yellow Macaws (Ara ararauna) – Oesophagus to Cloaca

Morphological studies of the gastrointestinal tract of blue‐and‐yellow macaws (Ara ararauna) are scarce. In view of the paucity of information regarding the digestive tract of macaws, this study aims to describe the gross anatomical features (oesophagus to cloaca) as part of a broad study of the gastrointestinal tract (GIT) of these birds. Three animals (two males and one female) adult macaws were anatomically dissected from the oropharynx to the cloaca to expose the GIT. The oesophagus was identified as a muscle‐membranous tube continuous with the crop, which was intimately attached to the skin. The internal longitudinal folds of the cervical oesophagus were sparser cranial to the crop and less evident compared to the portion caudal to the crop. The duodenum began in the pylorus and was grey‐coloured exhibiting a large lumen. The jejunum was formed by loops in a spiral‐fashion model supported by mesojejunum. The ileum was also composed by small loops and was continuous with the colo‐rectum forming the large intestine, because the caeca were absent. The large intestine was short, median in position, suspended in the dorsal wall of the abdominal cavity by mesentery and ended in the cloaca. The GIT was similar to the basic patterns in birds, in general, and also presented new unreported morphological data that might be important when studying nutrition and health of the macaws.     

Description of post‐implantation embryonic stages in European roe deer (Capreolus capreolus) after embryonic diapause

The embryonic stage of development is defined as the period between fertilization and the establishment of most of the organ systems by the end of this period. Development in this stage is rapid. In many mammalian species, particularly in humans, the interval between fertilization and implantation is exactly determined and continuous without intermission. However, European roe deer (Capreolus capreolus) embryos undergo a reversible retardation of development. This interesting reproduction strategy is called embryonic diapause (delayed implantation). After this period of embryonic arrest, development continues without further interruption. The aim of this study was to investigate embryonic development after diapause in European roe deer. Because of the embryonic diapause and the unknown date of fertilization, it was impossible to assign the embryos to a certain gestational age (days). This study describes normal stages of embryonic development mainly based on the external morphological traits of 56 well‐preserved post‐implantation roe deer embryos and attempts to assign the embryos to certain development stages. Carnegie stages of human embryos were used as an orientation for staging roe deer embryos. We observed a considerable range of variation of embryonic stages investigated until the end of January. We found post‐implantation stages of embryonic development already at the end of December and foetuses at the end of January. Moreover, assigning the embryos to a particular stage of development allows the comparison between pairs of twins and triplets. We showed that twins and triplets were always at the same development level, despite the discrepancy in inter‐twin and inter‐triplet size.     

Experimental Cryptosporidium infections in chickens: oocyst structure and tissue specificity

Oocysts of an avian isolate of Cryptosporidium were used to inoculate 21 chicks orally and 7 chicks intratracheally to determine the tissue specificity of this organism. Oocysts were passed in the feces 4 to 5 days after inoculation. Oocysts (6.8 by 5.0 microns) were fully sporulated and they were passed for at least 17 days by infected chicks. The mode of inoculation did not influence the distribution of cryptosporidia within the digestive tract. Cryptosporidia were found in the cloaca (100%), bursa of Fabricius (95.7%), terminal portion of the colon (26.1%), and cecum (4.3%) of chicks that were positive for developmental stages. Of 21 chicks inoculated orally, 4 had cryptosporidia in their trachea, whereas 6 of 7 chicks inoculated intratracheally had cryptosporidia in the trachea, bronchi, and air sacs. Cryptosporidium was found in the ducts of the salivary glands and nasal turbinates of chicks inoculated intratracheally that had clinical signs of respiratory tract disease. None of the chicks died or had intestinal disease.     

Recovery of Campylobacter from segments of the reproductive tract of broiler breeder hens

Three groups of >60-wk-old broiler breeder hens were assessed for the presence of Campylobacter within segments of the reproductive tract. In the first group, after stunned, the hens were bled, scalded, and defeathered, the reproductive tracts were aseptically excised from 18 hens, six from each of three adjacent floor pens that were feces positivefor Campylobacter. The reproductive tract segments (infundibulum, magnum-isthmus, shell gland, vagina, and cloaca) were pooled by pen. In the second group, 10 individual hens were sampled from the pens; the reproductive tract was divided into the following segments: magnum, isthmus, shell gland, vagina, and cloaca. For the third group, hens were obtained from two commercial farms that had been determined to be feces positive for Campylobacter, and the reproductive tract was divided into five segments, as described for the second group. Segments of the reproductive tract were placed into sterile plastic bags and suspended 1:3 (w/v) in Bolton enrichment broth, and serial dilutions were plated (0.1 ml) onto Campy-Cefex agar. The agar places were incubated at 42 C for 24 hr in a microaerobic atmosphere. In group 1, the pooled reproductive tract segments for hens from pen A were Campylobacter positive for the shell gland, vagina, and cloaca; hens from pen B were positive for the cloaca only; and hens from pen C were positive for the magnum-isthmus and doaca. In the second group, 9 of 10 cloaca samples were Campylobacter positive. Commercial hens in group 3 had campylobacter-positive cloaca samples (12/12), vagina (10/12), shell gland (7/12), isthmus (2/12), and magnum (4/12). Campylobacter colonization of the reproductive tract of the hen could enable vertical transmission of Campylobacter from the hen to the chick.     

Morphological changes of physeal cartilage and secondary ossification centres in the developing femur of the house mouse (Mus musculus): A micro‐CT based study

In mammals, long bones are formed by ossification of a cartilaginous mould during early stages of development, through the formation of structures called the primary ossification centre, the secondary ossification centres (SOCs) and the physeal cartilages (PCs). The PC is responsible for long bone growth. The morphology of the PC and the SOCs varies during different stages of femoral growth. In this respect, several details involving the process of murine femoral development are lacking. In the present study, a morphological characterization of femur development from the embryonic period to adulthood in mice was studied using micro‐computed tomography (micro‐CT). To achieve this aim, femora were collected at embryonic day (E) 14.5, E16.5 and E18.5 and at postnatal day (P)1, P7, P14, P35, P46 and P52. CT images were obtained using a micro‐CT scanner (X‐SkyScan 1172; Micro Photonics) and analysed using the micro‐CT 3D visualization software Mimics (Materialise NV, Leuven, Belgium) and NRecon (Micro Photonics). The results of the present study revealed that the femur and its PCs and SOCs undergo morphological changes during different stages of development, including changes in their shape as well as position and thickness. These changes may be due to the response of the femur to mechanical loads imposed by muscle surrounding the bone during these stages of development. The result of the present study is important to improve our knowledge related to ossification and growth patterns of mouse femur during development.     

Embryonic and fetal development in - pigmy rice rat - Oligoryzomys sp. (Rodentia, Sigmodontinae) and its significance for being a new experimental model

Oligoryzomys (Cricetidae, Sigmodontinae) is a common rodent genus from South America that includes a couple of very similar species. Related species have been used as experimental model for understanding several diseases for which these species are reservoirs. In order to provide a better understanding of the embryological aspects of this group, herein we showed data on the embryonic and fetal development in Oligoryzomys sp. Eight specimens of different stages of gestation were obtained from the Collection of the Zoology Museum of University of Sao Paulo, Brazil. Gestational ages were estimated by crown-rump-length according to Evans and Sack (1973). To address our analysis after examining the gross morphology, tissues from several organs were processed for light and scanning electron microscopy. Morphological data on the systems (nervous system, cardiorespiratory system, intestinal tract and urogenital system) were described in detail. Finally, the findings were compared with what is known about embryological aspects in other rodent species in order to establish similarities and differences during the organogenesis in different species.     

Expression of chicken LEAP-2 in the reproductive organs and embryos and in response to <Emphasis Type="Italic">Salmonella enterica</Emphasis> infection

In recent years host antimicrobial peptides and proteins have been recognised as key mediators of the innate immune response in many vertebrate species, providing the first line of defense against potential pathogens. In chickens a number of cationic antimicrobial peptides have been recently identified. However, although these peptides have been studied extensively in the avian gastrointestinal tract, little is known about their function in the chicken reproductive organs and embryos. Chicken Liver Expressed Antimicrobial Peptide-2 (cLEAP-2) has been previously reported to function in protecting birds against microbial attack. The aim of this study was to investigate the expression of cLEAP-2 gene in the chicken reproductive organs, as well as in chicken embryos during embryonic development, and to determine whether cLEAP-2 expression in the chicken reproductive organs was constitutive or induced as a response to Salmonella enteritidis infection. RNA was extracted from ovary, oviduct, testis and epididymis of sexually mature healthy and Salmonella infected birds, as well as from chicken embryos until day ten of embryonic development. Expression analysis data revealed that cLEAP-2 was expressed in the chicken ovary, testis and epididymis as well as in embryos during early embryonic development. Quantitative real-time PCR analysis revealed that cLEAP-2 expression was constitutive in the chicken epididymis, but was significantly up regulated in the chicken gonads, following Salmonella infection. In addition, expression of cLEAP-2 during chicken embryogenesis appeared to be developmentally regulated. These data provide evidence to suggest a key role of cLEAP-2 in the protection of the chicken reproductive organs and the developing embryos from Salmonella colonization.     

Prevalence of streptococcus bovis in racing pigeons

Summary

The prevalence of S. bovis in the intestinal tract of healthy racing pigeons was determined. Crop and cloaca swab samples obtained from 810 pigeons from 14 different lofts and from 122 pigeons that were presented for routine health control were examined for the presence of S. bovis. Pooled faecal samples were also obtained from pigeons in 82 different pigeon lofts. S. bovis was isolated from crop or cloaca samples of approximately 40 % of pigeons of all ages by direct culture and from 80 % of the pooled faecal samples by enrichment culture.

In a longitudinal study, crop and cloaca samples were collected every 3 months from pigeons in seven different pigeon lofts. The prevalence of S. bovis in these pigeons ranged from 0 to 100 %. The carriage rate was not related to the season or to the age of the pigeons.

The prevalence of S. bovis in organ lesions of pigeons examined at necropsy was investigated over a 35‐month period. S. bovis was isolated from 10 % of the birds examined. The incidence of S. bovis septicaemia was significantly higher in January to August than in September to December. It was concluded that S. bovis is an opportunistic pathogenic agent in pigeons.     

Gastrin and somatostatin enteroendocrine cells in the small intestines of ostrich (Struthio camelus var. domesticus) during pre -and post-hatching period

Studies on localization and distribution of enteroendocrine cells (EECs) are important for better understanding of their role in the ontogenetic development of intestines. Information about the distribution of the most important endocrine cells in the digestive tract of the ostrich is very limited; therefore, the aim of the present study was to identify gastrin and somatostatin EECs in the small intestine of the ostrich (Struthio camelus var. domesticus) chicks at different ages. Six embryos along with 42 ostriches of both sexes from hatching up to 60 days post-hatching, including six embryos, were obtained from an ostrich farm in Latvia. Duodenum, jejunum and ileum were investigated using routine histology and immunohistochemistry methods. Gastrin and somatostatin EECs were examined in 10 microscopic fields of the intestinal mucosa in each tissue sample. The cells were detected in all age groups as well as the embryos. The number of both types of EEC in the mucosa of the ileum was significantly lower (p < .01–.05) than in the duodenum. The present study suggested that the EEC may have a role in the development of the mucosa of the intestinal tract of ostriches with possible involvement in the development of the digestive functions.     

The development of the digestive tract in larval European catfish (Silurus glanis L.)

The European catfish, Silurus glanis L., has become an important aqua cultural fish in Croatia, and it is cultivated extensively in ponds in polyculture with carps. The development of the digestive tract in S. glanis was studied with the aim of improving intensive fish production. Research was carried out on S. glanis larval stadium from 1- to 19-day post-hatching (DPH). The main histological methods used were: haematoxylin and eosin staining, periodic acid Schiff staining (PAS), Alcian blue (AB) and toluidin blue staining (TB). A yolk sac was present during the first 5 days (1-5-DPH). During the initial 3-DPH period, there was no trace PAS and AB activity in the digestive tract. Differentiation of the digestive tract began at 3- to 5-DPH. The oesophagus was positive for AB at 5-DPH, PAS and TB after 7-DPH. Differentiation of enterocytes began at 5-DPG and the intestines were complete at 11-DPH. Development of liver and pancreas was also studied. The analysis of data obtained in this study suggests that after 5-DPH catfish larvae have morphologically completed digestive tracts.     

Thermosensitive period for sex determination of the tropical freshwater turtle Malayemys macrocephala

Many egg-laying reptiles possess temperature-dependent sex determination (TSD) in which outcome of gonadogenesis is determined by incubation temperature during a temperature-sensitive period of development. Prior studies on Malayemys macrocephala showed that incubation temperatures influence gonadal development and suggested that M. macrocephala exhibits TSD. However, information on the temperature-sensitivity period in this species was unknown until the current study. Turtle eggs were collected from rice fields in central Thailand from December 2016 to February 2017. In the laboratory, eggs were incubated at male-biased temperature (26 °C) and shifted to female-biased temperature (32 °C), or vice versa. Single shift experiments were performed systematically during embryonic stages 13–21. After hatching, sex of individual turtles was determined by histological analysis. We found that the sex determination of M. macrocephala is affected by temperature up to stage 16 of embryonic development.     

Seasonal changes in digestibility, passage rate and rumen fermentation of alfalfa hay in sika deer (Cervus nippon) under restricted feeding

To investigate seasonal variations in the digestive functions of sika deer, five female sika deer were provided with an amount of alfalfa hay cubes equivalent to voluntary food intake during winter. We measured the rate at which the food passed through the digestive tract, digestibility and rumen fermentation during the summer (August), autumn (November), winter (February) and spring (May). Total mean retention time in the digestive tract during summer and autumn was numerically longer than that in winter and spring, but the difference did not reach significance. Organic matter and fiber were less digestible in summer and autumn than in winter and spring (P < 0.05), whereas the digestibility of the dry matter tended to vary with the seasons (P < 0.1). Ruminal pH values seasonally changed (P < 0.01), and were the lowest in autumn. The concentration of ruminal ammonia‐nitrogen differed significantly among the seasons (P < 0.05), increasing in winter and decreasing during spring and summer. The numbers of protozoa changed significantly among the seasons (P < 0.05), being higher in autumn than in winter and spring, and intermediate in summer. The concentration of total volatile fatty acids was not seasonally affected, but the molar percentages of propionic acid and butyric acid significantly changed according to season (P < 0.05 and P < 0.01, respectively), and the ratio of acetic to propionic acid tended to change with the seasons (P < 0.1). The results of this study suggested that the digestive functions in sika deer, fed a commercial diet at a restricted level, differed notably among the seasons and these variations might partially be due to environmental effects.     

Anatomical parameters of the body and the digestive tract of Callithrix sp. marmosets under the influence of seasonality

Due to the ecological importance and given the scarcity of studies on the digestive morphology of primates, anatomical aspects of the body and digestive tract, as well as food content were analysed for marmosets Callithrix sp. (hybrids of exotic species) captured in forest fragments in Minas Gerais - Brazil, during the dry and rainy seasons, considering that seasonal variations affect the availability of food and quality of diet. Data such as body weight and length, and thoracic and abdominal perimeters were analysed, and no significant difference was found between dry and rainy seasons. In relation to the digestive tract, length, weight and diameter of the oesophagus, stomach, small intestine and large intestine were measured, in addition to the isolated caecum. There was a significant difference in the diameter of the ascending colon, which was larger in the animals in the dry season than in the rainy season. The difference found may be related to diet, and through analysis of the gastric and caecal contents, it was observed that gum was the main food item for the dry season marmosets, while in the rainy season the animals consumed mainly arthropods. Thus, the characteristic observed in the digestive tract of dry season marmosets is a reflection of an adaptive response to gum intake, since this food item is of low digestibility, necessitating that the food content be retained for a long time in this segment of the large intestine, helping the process of microbial fermentation and better energy utilization for these animals.     

STATIC AND DYNAMIC 18FDG‐PET IN NORMAL HISPANIOLAN AMAZON PARROTS (AMAZONA VENTRALIS)

Positron emission tomography (PET) is often used to stage and monitor human cancer and has recently been used in a similar fashion in veterinary medicine. The most commonly used radiopharmaceutical is 2‐Deoxy‐2‐[¹⁸F]‐Fluoro‐d ‐glucose (¹⁸F‐FDG), which is concentrated and trapped within cells that use glucose as their energy substrate. We characterized the normal distribution of ¹⁸F‐FDG in 10 healthy Hispaniolan Amazon parrots (Amazona ventralis) by performing whole body PET scans at steady state, 60 min after injection. Significant variability was found in the intestinal activity. Avian species are known to reflux fluid and electrolytes from their cloaca into their colon. To evaluate reflux as the cause of variability in intestinal distribution of ¹⁸F‐FDG, dynamic PET scans were performed on the coelomic cavity of six Hispaniolan Amazon parrots from time 0 to 60 min postinjection of radiotracer. Reflux of radioactive material from the cloaca into the colon occurred in all birds to varying degrees and occurred before 60 min. To evaluate the intestinal tract of clinical avian patients, dynamic scans must be performed starting immediately after injection so that increased radioactivity due to metabolism or hypermetabolic lesions such as cancer can be differentiated from increased radioactivity due to reflux of fluid from the cloaca.     

Oxygen Concentration and Cysteamine Supplementation During In vitro Production of Buffalo (Bubalus bubalis) Embryos Affect mRNA Expression of BCL‐2, BCL‐XL,MCL‐1, BAX and BID

This study examined the effects of O₂ concentration (5% vs 20%) during in vitro maturation (IVM), fertilization (IVF) and culture (IVC) or supplementation of IVM and IVC media with cysteamine (50 and 100 μm , respectively; IVM, IVF and IVC carried out in 20% O₂), on blastocyst rate and relative mRNA abundance of some apoptosis‐related genes measured by real‐time qPCR in immature and in vitro‐matured buffalo oocytes and in embryos at 2‐, 4‐, 8‐ to 16‐cell, morula and blastocyst stages. The blastocyst rate was significantly higher (p < 0.05) while the percentage of TUNEL‐positive cells was significantly lower (p < 0.05) under 5% O₂ than that under 20% O₂. The mRNA expression of anti‐apoptotic genes BCL‐2 and MCL‐1 was significantly higher (p < 0.05) and that of pro‐apoptotic genes BAX and BID was lower (p < 0.05) under 5% O₂ than that under 20% O₂ concentration at many embryonic stages. Following cysteamine supplementation, the blastocyst rate and the relative mRNA abundance of BCL‐XL and MCL‐1 was significantly higher (p < 0.05) and that of BAX but not BID was lower (p < 0.05) at many stages of embryonic development, although it did not affect the percentage of TUNEL positive cells in the blastocysts significantly. The mRNA expression pattern of these genes during embryonic development was different in 5% vs 20% O₂ groups and in cysteamine supplemented vs controls. At the 8‐ to 16‐cell stage, where developmental block occurs in buffalo, the relative mRNA abundance of BCL‐2 and MCL‐1 was highest under 5% O₂ concentration and that of BAX and BID was highest (p < 0.05) under 20% O₂ concentration. These results suggest that one of the mechanisms through which beneficial effects of low O₂ concentration and cysteamine supplementation are mediated during in vitro embryo production is through an increase in the expression of anti‐apoptotic and a decrease in the expression of pro‐apoptotic genes.     

Stereological analysis of metanephros from domestic cat (Felis catus,Linnaeus 1798) embryos and fetus

Kidney diseases are the most common illness for cats with a prevalence seven times higher than in dogs. Metanephros is the last of three renal systems to be formed during the embryonic period, which then becomes the permanent kidney. The current work aimed to analyse the morphology and to quantify the structures present in the development of metanephros from domestic cat (Felis catus) embryos and foetuses. For this purpose, the evaluation of the biometric parameters of metanephros from cat embryos and foetuses was performed in addition to the quantification of renal corpuscles and volume of cortical and medullary layers by stereological analysis. The evaluated biometric parameters were weight, width, height, thickness and volume. The values of the measured biometric parameters increased throughout the gestational stages. The quantity of renal corpuscles gradually increased following the embryo-foetal development, mainly during the middle of the gestational stage. It was during this phase that morphologically, a complete corticomedullary division was observed. Although the difference in the quantity of renal corpuscles between the middle and the end of the gestational stages was not statistically significant, there was an increase in the volume of the medullary layer and a decrease in the volume of the cortical layer between these two stages. These findings suggest that the metanephros presents a progressive growth with the renal corpuscles following this development until the middle of the gestational stage. Starting from this phase, the differentiation of the corticomedullary layers can be seen with a significant increase in the medullary layer.     

Developmental stages of the blue‐breasted quail (Coturnix chinensis)

Chickens and Japanese quail (Coturnix japonica) have traditionally been the primary avian models in developmental biology research. Recently, the blue‐breasted quail (Coturnix chinesis), the smallest species in the order Galliformes, has been proposed as an excellent candidate model in avian developmental studies owing to its precocious and prolific properties. However, data on the embryonic development of blue‐breasted quail are scarce. Here, we developed a normal developmental series for the blue‐breasted quail based on developmental features. The blue‐breasted quail embryos take 17 days to reach the hatching period at 37.7°C. We documented specific periods of incubation in which significant development occurred, and created a 39‐stage developmental series. The developmental series for the blue‐breasted quail was almost identical to that for chickens and Japanese quail in the earlier stages of development (stages 1–16). Our staging series is especially useful at later stages of development (stages 34–39) of blue‐breasted quail embryos as a major criterion of staging in this phase of development was the weight of embryos and the length of third toes.