The shell gland in laying and natural moulting commercial egg-type chickens: A histomorphological and ultrastructural study

The purpose of the present study was to determine the histological and ultrastructural changes in the luminal epithelium of the shell gland associated with natural moulting. Samples of the shell gland from laying (32 weeks old) and moulting (75 weeks old) hens were studied using histological, histochemical and electron microscopic techniques. In addition, TUNEL was used to demonstrate the distribution of apoptotic cells in the luminal epithelium of the shell gland. Autophagy, characterized by the presence of autophagosomes and autolysosomes, was evident in the early stages of degeneration in non-ciliated, ciliated and mitochondrial cells. The intermediate and advanced stages of regression in non-ciliated as well as mitochondrial cells occurred via apoptosis, while both apoptotic and necrotic ciliated cells were observed during the later stages of degeneration. The results of the present study suggest that a synergy of autophagy, apoptosis and necrosis is involved in the involution of the shell gland during natural moulting.     

Ultrastructure of the tubular glands in the isthmus region of the oviduct in laying and natural moulting commercial egg‐type chickens

The study investigated the ultrastructural characteristics of tubular gland and duct cells, as well as luminal gland cells in the isthmus region of the oviduct of laying and natural moulting hens. Tubular glands in laying birds were composed of type 1 and 2 cells. Based on the preponderance of each cell type, in relation to the location of a developing egg in the oviduct of the domestic fowl, these gland cells may represent different functional states of the same cell. The findings of the study on natural moulting birds suggest that autophagy is a process confined to the early stages of degeneration, while necrosis occurs in the terminal stages.     

Morphological features of the luminal surface of the magnum in the sexually immature ostrich (Struthio camelus)

Summary Observations were made, using scanning electron microscopy, of the surface features of the magnum in the immature ostrich during periods of ovarian inactivity, activity and regression. In birds with inactive ovaries the luminal surface of the magnum was lined with non-ciliated cells, which were densely covered by microvilli. In contrast, the magnum in birds with active ovaries was composed of ciliated and non-ciliated cells. The distribution of ciliated cells was not uniform, with clumps of cilia occurring next to non-ciliated areas. Samples collected from birds with regressing ovaries, during periods of decreasing daylength, revealed that the magnum was undergoing involution. The deciliation of ciliated cells and the presence of short microvilli on non-ciliated cells characterized magnal regression. These results suggest that ovarian activity and changes in daylength have a profound effect on the surface features of the magnum in the immature ostrich.     

Histological Characteristics of the Equine Oviductal Mucosa at Different Reproductive Stages

The objective of this work was to study cellular changes in the epithelium of the mare’s oviduct. Oviductal samples were taken from mares at different reproductive stages for optical microscopy and Hoechst 33258 staining. Glandular-like structures were observed in 100% of the oviducts. These structures were of the tubular type and were formed by ciliated and nonciliated epithelial cells arranged in a way similar to the epithelial surface. The amount of structures decreased progressively from the ampulla to the isthmus, but did not change through the different reproductive stages. Histological changes in the epithelium of the oviduct were observed associated with the reproductive stages. In the ampulla, the amount of ciliated cells decreased in the anovulatory phase compared with other reproductive stages. Cords of connective tissue lined by epithelium (trabeculae) and dividing completely the lumen of the oviduct were found in 50% of the oviducts. Epithelial cells projected toward the lumen as large vesicles of cytoplasm, sometimes containing a nucleus. The amount of cells presenting nuclear protrusion varied throughout the oviduct, with highest incidence in the ampulla, decreasing progressively toward the isthmus (P < .05). In addition, nuclear protrusions were higher in number during the anovulatory and luteal phases than in the other reproductive stages (P < .05). These nuclear protruding cells appeared to be extruding from the epithelium and showed no signs of apoptosis based on the histological and fluorescent stains used. The existence of these gland-like structures in the oviductal mucosa should be considered when studying the oviductal physiology in mares.     

Experimental bovine respiratory syncytial virus infection in conventional calves: ultrastructural respiratory lesions

The morphogenesis and repair of airway and alveolar injury induced by bovine respiratory syncytial virus (BRSV) was studied ultrastructurally in conventional calves to characterize pulmonary cell types susceptible to viral infection and cytopathologic changes associated with infection. Viral nucleocapsids and budding virions were present in tracheal and bronchial ciliated and nonciliated epithelial cells and mucous cells 3, 5, and 7 days after inoculation and in bronchiolar ciliated and nonciliated epithelial cells 5 days after inoculation. Mild interstitial pneumonia was observed 5 days after inoculation and was characterized by swelling of type 1 and type 2 alveolar epithelial cells, interstitial edema, and infiltration by lymphocytes and macrophages. Viral assembly and release in tracheal and bronchial epithelial cells was associated with loss of cilia from ciliated cells, formation of syncytial epithelial cells, swelling of mitochondria and endoplasmic reticulum, and cell necrosis. Neutrophils, lymphocytes, and macrophages were present in close association with the viral-infected and damaged epithelial cells. There was intercurrent hyperplasia of basal epithelial cells that, in association with other epithelial lesions, resulted in the loss of normal ciliated epithelium in these airways 5 and 7 days after inoculation. Regeneration of airway epithelium was largely completed by 10 days after inoculation, except in 1 of 4 calves that had failure of epithelial repair and that developed secondary bacterial pneumonia. Pulmonary ultrastructure in BRSV-inoculated calves 30 days after inoculation was indistinguishable from that in controls. The results demonstrated that BRSV can induce reversible alterations in airway epithelium, which may cause depression of mucociliary clearance and thereby enhance susceptibility to bacterial infection.     

Evaluation of cytopathologic changes induced in chicken tracheal epithelium by Mycoplasma gallisepticum in vivo and in vitro

Changes in tracheal epithelial surfaces induced by Mycoplasma infection in vivo and in vitro included release of mucous granules followed by exfoliation of ciliated and nonciliated epithelial cells. Light microscopy, scanning electron microscopy, and transmission electron microscopy confirmed that the loss of cilia from individual cells was infrequent. Epithelial cells typically lost their intercellular connections, rounded up, exfoliated, and then lysed--giving rise to a population of cellular organelles, such as mitochondria and cilia intermixed with mucus to form the exudate found within the tracheal lumen. Repair of the epithelial surface was effected by basilar epithelial cells differentiating and filling in the spaces formed by exfoliated cells. These cells were hypertrophied, nonciliated at 14 days after infection in vivo, and covered with microvilli. In sectioned material obtained during the infection, there was increasing epithelial thickness due to cellular infiltration and edema. Tracheal rings in vitro showed similar changes to those seen in vivo, except that exfoliation was more severe and occurred earlier. In addition, there were no cellular infiltration due to the lack of a vascular supply and only a small amount of mucus due to the smaller number of mucous cells available to release into the tracheal lumen.     

Interaction of Tritrichomonas foetus and the bovine oviduct in an organ culture model

Tritrichomonas foetus is an extracellular parasite of the reproductive tract in cattle. The mechanism by which T. foetus causes abortion in cattle is largely unknown. There are no studies of infection in the cow oviducts, almost all published papers are related to vagina infection and few articles focusing on the uterus. The aim of the present study was to establish a working model of bovine oviduct epithelial cells and submit these cells to Tritrichomonas foetus interaction. Twenty bovine oviducts were obtained from cows at a commercial abattoir and T. foetus was injected through the isthmus into the oviduct lumen. The whole oviduct was analyzed by scanning and transmission electron microscopy. The results reported here demonstrate that: (1) fresh whole oviducts can be used as a good model to study parasite-host cell interaction; (2) cow oviduct epithelium has been shown to consist of two cell types: ciliated and nonciliated secretory cells, and T. foetus displayed great specificity for the nonciliated cells localized in the deeper oviduct folds; (3) T. foetus adheres as single separate cells, and maintains the flagella externalized; (4) differently from T. vaginalis, T. foetus does not change its shape during the adhesion process; and (5) oviduct cells exhibited morphological characteristics of apoptosis after trichomonadal interaction.     

Localisation of MHC class II,lymphocytes and immunoglobulins in the oviduct of laying and moulting hens

1. Our aim was to determine the presence and numbers of immunocompetent cells in the oviduct of laying and moulting hens. Immunocompetent cells were localised by immunocytochemistry throughout the entire length of the oviduct.

2. In laying birds, MHC class II + cells were observed in the subepithelial and middle part of the stroma of all oviducal segments and the mucosal epithelium of the infundibulum and vagina. CD3 + cells were also localised in subepithelial and middle part of stroma as well as in mucosal epithelium of each oviducal segment. Bu‐lb + and IgG + cells were also observed in the epithelium and subepithelial and middle part of the stroma of all oviducal segments, though stroma of the magnum, isthmus and uterus contained few Bu‐lb + cells. IgA + cells were observed only in the mucosal epithelium of the magnum in small numbers.

3. In moulting hens, there were few numbers of immunocompetent cells in the mucosal epithelium of each oviducal segment, although CD3 + cells were observed in the infundibulum and vagina. In the subepithelial stroma, the populations of MHC class II + cells in the infundibulum, magnum and uterus, CD3 + cells in the infundibulum and vagina, as well as IgG + cells in each oviducal segment except for isthmus were smaller than in laying hens. In contrast, the number of immunocompetent cells in the middle part of stroma of moulting hens were equal to or greater than in laying hens.

4. These results suggest that the oviducal immune function is active in the surface tissues of the mucosa in laying hens, whereas it is reduced in moulting hens.     

Morphometric and ultrastructural changes in ciliated cells of the oviductal epithelium in prolific Chinese Meishan and Large White pigs during the oestrous cycle.

There are some differences in reproductive features between the Chinese Meishan (CM) pig and Large White (LW) pig. The aim of the present study was to investigate the quantitative changes and ultrastructural features of ciliated cells in the various regions of the CM and LW pig oviduct during the follicular and luteal phases of the oestrous cycle. In the fimbrial and ampullar epithelia at the follicular phase, the ciliated cells were more plentiful than in the isthmic region in both pigs. In the CM pigs, there was a striking decrease in the percentage and cell height of ciliated cells in the luteal phase compared with the follicular phase. Although similar quantitative changes were observed in the LW pig oviduct, these changes were less dramatic than that in the CM pig oviduct. In both pigs, the percentage and cell height of ciliated cells in the isthmus were unchanged between the follicular and luteal phases. The ultrastructure of ciliogenic and ciliated cells was observed. In the fimbrial and ampullar epithelia during the follicular phase, most of the ciliated cells showed normal morphology, having many elongated cilia and mitochondria, but in the CM pig oviduct during the luteal phase many of the ciliated cells possessed immature cilia and swollen mitochondria. Cells undergoing ciliogenesis were frequently observed in the fimbriae and ampulla, and occasionally in the isthmus. Cytoplasmic protrusions containing variable numbers of ciliary axonemal complexes occurred in the fimbrial and ampullar epithelium in the CM pigs at the luteal phase, suggesting that deciliation of cells occurs by membrane-bound cilia packets forming at the apices of cells and pinching off. These results demonstrate that there are regional variations in the cyclic changes associated with the oviductal ciliated cells of the pigs, while there are marked morphometrical and ultrastructural changes in oviductal ciliated cells of the CM pigs compared with that of the LW pigs.     

Ultrastructural pathology of Bordetella avium infection in turkeys

One-day-old turkeys were infected intranasally with Bordetella avium, and tracheas were examined by scanning and transmission electron microscopy at 1 to 5 weeks post-inoculation (PI). The predominant ultrastructural lesions were progressive loss of ciliated epithelium with replacement by nonciliated cells, bacterial colonization of ciliated cells, membrane-bound crystalline inclusions in cytoplasma of epithelial cells, depletion of mucous granules, and distortion of tracheal rings and the mucosal surface. Tracheal surface exudates consisted of mucus, necrotic cells, heterophils, and fibrin. Ciliated cells were replaced by immature cuboidal cells characterized by abundant rough endoplasmic reticulum with small numbers of electron-dense mucous granules in the apical cytoplasm. Bacterial surfaces were rough and contained numerous pleomorphic, knob-like structures, 20-50 nm in diameter. Other changes included enlarged mucosal gland openings, cell extrusion marks, pleomorphic microvilli, and cells with small numbers of short cilia.     

Light and Scanning Electron Microscopic Studies of Oviductal Epithelium in Thai Swamp Buffalo (Bubalus bubalis) at the Follicular and Luteal Phases

The purpose of this study was to investigate the morphological changes in the epithelium of Thai swamp buffalo oviducts at the follicular and luteal phases by histological technique and scanning electron microscopy. The samples from the infundibulum, ampulla, isthmus and uterotubal junction (UTJ) of the oviduct were taken immediately after slaughter at the local abattoir. Noticeable cyclic changes were observed on the epithelial surface of the infundibulum and ampulla, but few changes were present in the isthmus and UTJ. At the follicular phase, the epithelium of infundibulum and ampulla were densely covered with ciliated cells whose cilia concealed the apical processes of the secretory cells. In contrast, the secretory cells dominated in the epithelium at the luteal phase and most of the ciliated cells were hidden by the bulbous processes of these cells. In the isthmus and UTJ at the follicular and luteal phases, the secretory cells were almost flat or gently rounded and covered with numerous microvilli at their apical surface. In conclusion, the histological and ultrastructural observation of Thai swamp oviduct epithelium revealed marked cyclic changes in the cellular differences associated with the main functions of segmental variations.     

Prenatal differentiation of bovine oviductal epithelium: an electron microscopic study

In this study, we investigated the ultrastructural changes during the prenatal differentiation of oviductal epithelium in 16 bovine embryos and fetuses from CRL of 18.0 cm to a CRL of 94.0 cm. Ciliated and secretory cells of bovine uterine tube, a derivative of the Müllerian duct, differentiate to distinct development stages in the prenatal period. The typical cellular pattern, which is generally characteristic for the adult bovine oviduct, is also obtained during fetal life. In the early stages (CRL 18.0/20.4 cm), the bovine oviductal epithelium appears mostly undifferentiated. The epithelial cells show only a few mitochondria, some cisternae of rough endoplasmic reticulum (rER) and a small Golgi-complex. Most of the cytoplasm is filled with a large amount of glycogen, which decreases during later development. Interspersed between the undifferentiated epithelial cells, a few cells undergoing ciliogenesis can be observed. Ciliogenesis increased significantly during the later prenatal developmental stages. At a CRL of 55.0 cm, ciliated cells appear fully differentiated with mature cells covering their luminal surface. Formation of cilia usually use the acentriolar pathway. Fibrous granules occurred initially in association with the Golgi-apparatus and r(ER) in the supranuclear cytoplasm. Fibrous granules later fuse with deuterosomes and give rise to procentrioles, which are translocated to the luminal plasma membrane. There they become arranged in a line just beneath the apical cell membrane and further differentiate to basal bodies from which the formation of cilia and striated rootlets take place. Clear signs of differentiation of secretory cells were first seen in our material in fetuses with a CRL of 51.0 cm and 64.0 cm. These cells contain a well developed rER and Golgi-apparatus with dilated cisterns. In the supranuclear cytoplasm, the number of secretory granules continuously increases during later development and the cells adapt to the morphology of mature secretory cells at the CRL 94.0 cm.     

Ultrastructural features of the uterus in the sexually immature ostrich (Struthio camelus) during periods of ovarian inactivity and activity

The ultrastructure of the surface epithelium and tubular glands of the uterus in the immature ostrich is described. In ostriches with inactive ovaries the uterus is lined by a non-ciliated simple columnar epithelium, with basally located heterochromatic nuclei. Scanning electron microscopy revealed that these non-ciliated cells have a dense microvillous cover. A simple columnar to pseudostratified columnar epithelium, comprised of non-ciliated and ciliated cells, lines the uterus in birds with active ovaries. The ciliated cells possess a wide luminal region, which contains a nucleus and various organelles. An accumulation of secretory granules was observed in the apical regions of the non-ciliated cells, as well as in a few ciliated cells. In addition to non-ciliated and ciliated cells, a cell type with rarefied cytoplasm was also identified. These cells appear to correspond to calcium secreting cells identified in other avian species. The results of this study indicate that, although uterine differentiation is present in immature ostriches with active ovaries, the production of secretory product appears to occur mainly in non-ciliated epithelial cells.     

Anatomicohistological Characteristics of the Tubular Genital Organs of the Female Red Brocket Deer (Mazama americana) in the Peruvian Amazon

This study examined the anatomical and histological characteristics of tubular genital organs of 51 adult female red brocket deer in the wild in different reproductive stages, collected by rural hunters in the north‐eastern Peruvian Amazon. The infundibulum was characterized by a large diameter and the presence of a highly folded and ciliated epithelium, and the isthmus has a growing secretor epithelium and a thicker muscular layer. Whereas ciliated cells are more frequent in the infundibulum, epithelial secretory cells showing abundant apical secretory blebs are more frequent in the isthmus. In non‐pregnant females in luteal phase, the endometrium transforms from a proliferative to a secretory type, showing a significant proliferation of endometrial uterine glands. The red brocket deer has four large circular folds in the cervix. The epithelium of the cervix is composed primarily of secretory cells. In pregnant females, the lumen of the endocervical canal is occupied by abundant mucous secretion. All pregnant females had one embryo or fetus, with a fetal sex ratio of 54.0% females to 46.0% males. This species has a cotyledonary, syndesmochorial and partially deciduate placenta, with 6–7 dome‐shaped caruncles per female. The red brocket deer does not present a true cornification of the vaginal epithelial cells, and no vaginal epithelial pattern was determined according the reproductive state of the female.     

Macroscopic and microscopic anatomy of the oviduct in the sexually mature rhea (Rhea americana)

The morphological characteristics of the oviduct of 12 sexually mature rheas (Rhea americana) were studied. Only the left oviduct is developed as a long tube with a length of 122 +/- 23.1 cm, and is subdivided into infundibulum (15.2 +/- 4.0 cm), magnum (63.3 +/- 9.4 cm), isthmus (5.6 +/- 3.1 cm), uterus (16.0 +/- 4.2 cm) and vagina (11.5 +/- 1.4 cm). The mucous membrane of the oviduct, as a whole, possesses luminal folds covered by ciliated columnar epithelium with secretory cells. The infundibulum part presents a cranial opening with thin and long fimbriae with few tubular glands in caudal tubular portion. In the magnum, the largest portion of the oviduct, the folds are thicker and are filled with tubular glands. The isthmus is short and presents less bulky folds and a few tubular glands. A bag-shaped uterus in the cranial area shows thin folds, and in the caudal region (shell gland) more ramified folds with few tubular glands. The vagina has long luminal folds and a thick muscular tunic; no glands with sperm-storage characteristics have been observed. In conclusion, the oviduct in sexually mature rhea has morphological similarities with the other species of birds already described; however it presents its own characteristics to produce a big egg.     

Scanning electron microscopic study of the lower respiratory tract in calves and adult cattle

The surface characteristics of the lower respiratory tract of two groups of cattle were studied with the scanning electron microscope. Group A comprised six one-week-old calves and group B four adult cows. None of the animals had overt respiratory disease or gross morphological evidence of pulmonary lesions. The trachea, bronchi, bronchioles and alveoli of the cranial and the caudal lobes of the right lung were examined. In both groups the luminal surface of the trachea and large bronchi were completely covered by cilia, apparently forming an efficient mucociliary escalator. In the adult animals there were some patchy areas in the trachea and large bronchi devoid of ciliated cells; these were considered abnormal. In the bronchi, non-ciliated cells, mainly mucus-secreting, were not easily identified unless they were discharging secretion. In small bronchi, non-ciliated cells were more evident and easily seen. The bronchioles had many non-ciliated cells and very few ciliated cells capable of forming a complete carpet for a mucociliary escalator. Types 1 and 2 alveolar epithelial cells and alveolar macrophages were identified in both groups. Pores of Kohn were found in the alveolar walls in all animals.     

Ultrastructural demonstration of cilia and ciliary rootlets in mammalian uterine tube epithelium in different functional states.

Tissue from the infundibulum region of the uterine tube (oviduct) of guinea pigs, cattle, sheep, and swine was examined by electron microscopy. In all specimens, cilia and ciliary rootlets were present in variable numbers in the ciliated cells during both the follicular and the luteal phases of the estrous cycle. True degeneration of cilia was not evident during luteal phase or pregnancy. The ciliary rootlets in ruminant and nonruminant species had structural similarities to those described in people and rhesus monkeys. These organelles measured approximately 1 mum long in most species studied, but in the cow, the rootlets, extending downward into the cytoplasm from the proximal end of the basal body, reached a length of 2 mum. The rootlets had a cross-striation of thick and thin bands, the period measuring 40 to 60 nm. Mitochondria were closely associated with the rootlets. The rootlets usually formed a small angle to the axis of the cilium. These organelles probably function as anchoring or stabilizing structures for the motile cilia. Results indicated that the rootlets are more widely distributed in mammalian uterine tube cilia than previously postulated. Polyribosomes, microfilaments, microtubules, and electron-opaque fibrous granules were frequently seen in the cytoplasm of the ciliated cells. The presence of fibrous granules in close association with the basal bodies indicate that these granules have a role in the development of cilia and rootlets. Cilia and precursor fibrous granules were also seen in porcine fetal uterine tube epithelial cells.     

An Immunohistochemical Study of the Oviduct in the Domestic Fowl (Gallus domesticus)

This study describes the distribution of vimentin, desmin, smooth muscle actin (SMA) and laminin in the oviduct of the laying domestic fowl. Vimentin immunostaining was localised in the luminal epithelium of the infundibulum, magnum, magnum–isthmus junction and isthmus. The luminal epithelium of the shell gland regions displayed weak vimentin immunostaining. Vimentin immunostaining was demonstrated in the glandular grooves of the tubular infundibular region. In contrast, gland cells in the magnum, isthmus and shell gland regions were vimentin immunonegative. Fibroblasts and vascular endothelial cells in the lamina propria of the oviductal regions studied exhibited vimentin immunostaining. Strong desmin and SMA immunostaining were present in the smooth muscle cells of the tunica muscularis and vascular tunica media. In this study, basement membranes underlying the luminal and glandular epithelia were immunopositive for laminin. In addition, basement membranes associated with smooth muscle cells exhibited laminin immunostaining. The results of the study indicate that the immunolocalisation of desmin, SMA and laminin in the oviduct of the domestic fowl is similar to that in the mammalian uterus. The immunolocalisation of vimentin in the domestic fowl varies depending on the oviductal region.     

The surface features of the epithelial lining of the ducts of the epididymis of the ostrich (Struthio camelus)

The luminal appearance of the various ducts of the epididymis of the ostrich was studied by scanning electron microscopy in tissues fixed by immersion in glutaraldehyde. The ductal types were similar to those previously described for some other species of birds. Numerous short microvilli, as well as a single cilium, projected from the apical surface of the rete testis cell. The ciliated cells of the efferent ductules projected tufts of cilia into the ductal lumen, while the non-ciliated cells bore short microvilli. The connecting and epididymal ducts were lined by a columnar cell type whose apical surface bore uniformly distributed microvilli and a single, centrally situated cilium. The spermatozoa found in all ducts of the epididymis bore a distal cytoplasmic droplet. This observation has implications for the maturational process in the ostrich spermatozoon in the epididymis. The surface features of the ducts, except for a few noteworthy differences, were generally similar to those previously described for the male domestic fowl, turkey and duck.     

Pulmonary lesions induced by 3-methylindole and bovine respiratory syncytial virus in calves

Our objectives were to describe the ultrastructural morphogenesis of pulmonary lesions induced by 3-methylindole in 30- to 45-day-old Holstein calves and to determine whether toxic exposure to 3-methylindole exacerbates pulmonary lesions induced by bovine respiratory syncytial virus. Administration of 3-methylindole (0.25 g/kg) to calves resulted in interstitial edema and ultrastructural swelling of type-I alveolar epithelial cells and nonciliated bronchiolar epithelial cells as early as 4 to 6 hours after intraruminal administration. More severe alveolar edema containing protein was associated with swelling of capillary endothelial cells at 2 days after administration. Proliferation of type-II alveolar epithelial cells was first observed at 2 days after 3-methylindole administration, and marked hyperplasia of type-II epithelial cells and nonciliated bronchiolar epithelial cells was evident by 4 days after administration. Pulmonary cytochrome P-450 monooxygenase concentrations decreased significantly (P less than 0.001) by 12 hours after administration and did not increase significantly again by 8 days after administration. Calves were inoculated with bovine respiratory syncytial virus 3 days after administration of 3-methylindole, and pulmonary lesions were assessed 5 days after viral inoculation. Viral replication was demonstrated by fluorescence microscopy for viral antigen or by transmission electron microscopy in ciliated and nonciliated airway epithelial cells. Viral antigen was identified infrequently in alveolar macrophages and in type-II alveolar epithelial cells. 3-Methylindole exposure in calves did not result in more widespread distribution of viral antigen in alveolar tissue of respiratory syncytial virus-inoculated calves or in significant enhancement of viral pneumonia.(ABSTRACT TRUNCATED AT 250 WORDS)