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The biological effects of simulated photoperiod and melatonin on the control of reproduction of guinea fowls (Numida meleagris) are not well understood. Herein, thirty (30) sexually mature guinea fowl cocks were randomly assigned to 1-6 groups (n = 5) and subjected to different photoperiodic regimes in the presence or absence of exogenous melatonin (Mel; 1 mg/kgBW/day, i/m) for eight weeks. Testes of the euthanized cocks were processed for gross morphology, histological, histochemical, and oxidative stress markers. Testosterone concentration was determined in serum samples using the enzyme-linked immunosorbent assay (ELISA) technique. We observed an increase in testicular size in the Mel and Non-Mel groups under long-day (LD) photoperiods, and in the Non-Mel group under short-day (SD) photoperiod. Conversely, the testicular size was drastically reduced in the Mel group for SD. Seminiferous tubules in the Mel and Non-Mel groups of the SD showed cytomorphological changes, including degenerated cells, focal vacuolations, and depletion of germinal epithelium. However, the germinal epithelium appeared to be complete and active in both the Mel and Non-Mel groups for the LD. In all groups, the testes showed positive staining for PAS with varying intensities. There was a significant difference in PAS-staining intensity between different photoperiodic regimes and exogenous melatonin. The study observed the interaction between photoperiods and exogenous melatonin on glutathione reductase (GSH), malondialdehyde (MDA), and serum testosterone. Overall, the results indicated that a long-day (LD) photoperiod, combined with exogenous melatonin, enhanced reproductive activity in male guinea fowl by increasing testicular size and serum testosterone concentration., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The purpose of this study is to describe, in detail, the ultrastructure of the infundibulum of the sexually mature and active female green iguana, Iguana iguana. The infundibulum of five iguanas was remarkably distinct from the uterus, and was also clearly demarcated into cranial (expanded v-shaped) and caudal (tubular) divisions. Tissue samples obtained from five portions (three from the cranial division and two from the caudal division) of the infundibulum were processed conventionally for light and electron microscopy. The epithelial lining of the most anterior, middle, and posterior, parts of the cranial division displayed nonciliated cells predominantly, and occasionally ciliated cells. The numerous secretory granules in nonciliated type 1 cell found in the fimbrial aspect of the infundibulum were homogenous and deeply electron-dense, but those in the other two regions were variants of this cell type because they contained variably electron-dense secretory granules. Two main types of nonciliated cells (type 2 and its variant, type 3, as well as type 4) occurred in the epithelial lining of the caudal division of the infundibulum, but they, clearly, showed no dense secretory granules. Whereas the nonciliated type 2 cell and its variant (type 3 cell) contained large glycogen deposits, the type 4 cell lacked these deposits but its apical part contained large lipid-like droplets and, remarkably, blebbed into the duct lumen. The nonciliated cells lining the mucosal tubular glands contained highly electron-dense secretory granules, which were similar to those found in the nonciliated type 1 cell in the epithelial lining of the fimbrial part of the cranial division of the infundibulum., (© 2023 Wiley Periodicals LLC.)

Phthalate esters, such as di(n-butyl) phthalate, (DBP), are synthetic chemical pollutants commonly used as plasticizers in the manufacture of plastics. In the present study, we investigated the effects of DBP in the testes of adult male quails (Coturnix cortunix japonica) exposed by oral gavage to variable doses of DBP (0 [control], 1, 10, 50, 200, and 400 mg/kgbw -d ), for 30 days during the prepubertal period, using histo-morphometric and ultrastructural techniques. Generally, significant decreases in seminiferous tubular diameter (STD) and epithelial height (SEH) were observed predominantly at the highest DBP doses (200 and 400 mg/kg), as compared to medium (50 mg/kg), and lowest doses (1 and 10 mg/kg) as well as the control group. Ultrastructurally, apparent dose-specific degenerative changes were observed in the Leydig cells. The lowest DBP doses (1 and 10 mg/kg) did not produce significant effects on Leydig cell ultrastructure, whereas, at the highest doses (200 and 400 mg/kg), the Leydig cells were remarkably conspicuous in the interstitium and appeared foamy. There was a preponderance of electron-lucent lipid droplets which crowded out the normal organelles of the cell, as well as increases in the number of dense bodies in the cytoplasm. The smooth endoplasmic reticulum (sER) was less obvious, compacted, and wedged between the abundant lipid droplets and mitochondria. Taken together, these findings indicate that pre-pubertal exposure of precocious quail birds to DBP, produced parameter-specific histometric tubular changes, as well as dose-dependent cyto-structural derangement of the Leydig cells; which consequently may lead to overt reproductive impairments in the adult bird in the environment., (© 2023. The Author(s).)

Avian species comprise more than half of all vertebrates yet there is a dearth of information regarding spermatid development in this class of animals. This report of spermiogenesis in the cattle egret, Bubulcus ibis, is the first in the order Pelecaniformes. Five sexually mature and reproductively active male cattle egrets were captured in the wild, humanely euthanized, the reproductive organs dissected out, and tissues from the testes routinely prepared for transmission electron microscopy. Twelve steps of spermatid development, using the step-wise system, were determined. Acrosomogenesis in the egret results in a relatively short, solid, bullet-shaped acrosome that ends bluntly anteriorly and flat posteriorly or basally. The nucleus displays remarkable morphological changes, with the anterior end of the mature spermatid becoming flat, lacking a rostrum and an endonuclear canal. A perforatorium does not develop. It is noteworthy that a longitudinal, but not a circular, manchette develops during spermiogenesis in this bird. The proximal centriole is attached to the nucleus, at the implantation fossa, by means of well-formed, electron dense struts of material. An amorphous fibrous sheath develops in the principal piece. The interesting development and peculiar features of the acrosome and nucleus, as well as the absence of the perforatorium and circular manchette in the spermatozoon of the cattle egret, may be of phylogenetic significance., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

It remains a major concern that sperm structure has continued to be poorly investigated and reported in avian species. To our knowledge, sperm structure in the order Pelecaniforme has not been reported. Although McFarlane (1963; Proceedings of the XIII International Ornithological Congress; Ithaca, NY; American Ornithologists' Union) reported the study of spermatozoa in two genera and two species of the family Ardeidae, he did not provide an account, or the names of the species examined. The present report on the sperm structure of the cattle egret, Bubulcus ibis, is, thus, the first in the order Pelecaniformes (this bird has been placed variably under the order Ciconiiformes, or the order Pelecaniformes). Five sexually mature and reproductively active male cattle egrets were obtained from the wild, humanely euthanized, the reproductive organs dissected out, and tissues from the ducti deferentia were prepared for transmission electron microscopy. The sperm structure of this bird is generally similar to that described for most non-passerine birds. However, the acrosome is a short, conical or bullet-shaped, blunt-ending organelle that lacks a perforatorium. The base of the acrosome is flat and makes contact with the nucleus along, a correspondingly flat plane. The nucleus, thus, ends anteriorly in a flat plane devoid of a concavity or a rostrum, and an endonuclear canal. The acrosomal and nuclear features of this bird are, therefore, main deviations from the situation in the non-passerine clade of birds., (© 2020 Wiley-VCH GmbH.)

Only a few studies on the development of the passerine spermatozoon are available, yet species variations in the conformation as well as structure of the generally helical acrosome have been reported. This study of spermiogenesis in the Carib grackle (Quiscalus lugubris) intended to provide a deeper understanding of the development of the sperm, and in particular to investigate the bi-partite nature and development of the acrosome as well as its relationship with the nucleus, in the absence of a perforatorium that is found in most non-passerine birds. The acrosomal vesicle already displays a bi-partite nature in the acrosomal granule within the Golgi complex, and the attachment of the dense granule (future acrosomal core) within the crest part (future acrosomal crest) establishes polarity as it approaches and attaches to the nucleus. Thereafter, they develop variably. The acrosomal crest leads the elongation and spiraling of the acrosome, and the core portion contributes significantly to the formation of the keel of the crest part. The rounded, core-bearing part of the base of the acrosome progressively indents and fits into the concavity, thus formed, at the anterior part of the nucleus. The possible homology of the acrosomal complex (including the perforatorium) and the nucleus between non-passerine and passerine birds was discussed. The centriolar complex comprises both the proximal and distal centrioles in all spermatids and spermatozoa. The mitochondria undergo a number of morphological changes, including size and electron-density, from the round spermatid through to the mature spermatid; changes that are probably influenced by their functional states in the different evolving phases of the spermatids., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The spermatozoon of the Carib grackle, Quiscalus lugubris, a member of the family Icteridae, is generally similar in organization to the passerine-type of spermatozoon, in being highly elongated and displaying a helical structure of the acrosome, nucleus and principal piece of the tail. There are subtle variations in acrosomal structural features between this organelle in the grackle and that in some of the very few passerine species of birds in which the spermatozoon has been studied. The proximal centriole is present, and, thus, the Carib grackle is the third passeridan bird in which this organelle, hitherto regarded as absent in passerine birds, has been described in the spermatozoon. The spermatozoon of this bird also possesses a granular helix, which feature has been found variably even in the scanty available reports on passerine spermatozoa. It is advocated that the spermatozoon be studied in many more species of this large clade of birds. This report provides a basis for the study of spermiogenesis in the Carib grackle, with the aim of exposing, inter alia, a number of developmental features and processes of certain organelles that have received attention, recently, in the spermatozoa of passerine birds., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In the present study, we have investigated the effects of 30-day dietary (pre-pubertal) exposure to different doses (0 (control), 1, 10, 50, 200 and 400 mg/kg bodyweight/day) of di(n-butyl) phthalate (DBP) on Leydig cells of adult male Japanese quails by quantifying the transcript levels for P450 side-chain cleavage (p450scc), P450c17 (CYP17), and 3β- and 17β-hydroxysteroid dehydrogenase (hsd) using quantitative (real-time) polymerase chain reaction (qRT-PCR). In addition, the plasma testosterone levels were analysed using radioimmunoassay (RIA) and testis was examined for evidence of gross pathology and histopathology. Our data showed that pre-pubertal exposure to DBP produced alterations in testicular architecture as evident by poorly developed or mis-shaped testis, and altered spermatogenesis due to tubular degeneration and atrophy of seminiferous tubules especially in the high DBP dose (200 and 400 mg/kg) treated groups. In addition, DBP altered several key enzymes involved in testicular steroidogenesis pathways in an apparent dose-dependent manner. For example, biphasic effects of DBP were observed for P450scc and 3β-hsd mRNA, that were generally increasing at low dose 10 mg/kg, and thereafter, an apparent dose-dependent decrease between 50 and 400mg/kg. The steroidogenic acute regulatory (StAR) protein was at the lowest detectable limits and therefore not quantifiable. These effects did not parallel the non-significant changes observed for plasma testosterone levels. The present data is consistent with previous reports showing that DBP modulates Leydig cell steroidogenesis in several species, with a potential negative effect on reproduction in those avian species that are vulnerable to endocrine disrupting chemicals., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Current knowledge on avian spermiogenesis, including strengths and weaknesses, has been reviewed. Information on avian spermiogenesis considerably lags behind that in mammals because of the paucity of reports in birds. Spermiogenesis in passerine birds has received even much less attention than in non-passerine birds. Mechanisms underlying morphogenesis of the acrosome and nucleus, and roles of microtubular assemblies are poorly understood. The proximal centriole found in non-passerine birds, but hitherto considered to be absent in passerine birds, has recently been described in spermatids and mature spermatozoa of 2 passeridan species, including the Masked weaver for which new and detailed spermiogenetic information is provided in this review. A great deal more studies on spermiogenesis, and spermatogenesis generally, in various avian species are required to considerably enhance knowledge of this phenomenon, contribute to comparative spermatology, provide a basis for appropriate applied studies, and contribute to understanding of phylogeny in this vast order of vertebrates.

Simple testicular cysts are rare in birds and mammals. However, the condition has recently been reported in the ostrich (Struthio camelus), an economically important farmed bird. The innervation of normal and cryptorchid testes, unlike the simple testicular cysts of birds and mammals, has received considerable attention. This study, therefore, immunohistochemically demonstrates the presence and the general distribution pattern of neuronal and glial markers in the simple intratesticular cyst, and its associated structures, of fourteen adult ostriches using antibodies to neurofilament protein, S-100, neuron-specific enolase and protein gene product 9.5. The LSAB+ Kit(DakoCytomation, Denmark) immunostaining protocol was used in this study. The normal seminiferous peritubular tissue showed few or no immunoreactive nerve fibres. A greater density of neurofilament protein, S-100, neuron-specific enolase and protein gene product 9.5 immunopositive nerve fibres were observed in the tunica albuginea adjacent to the cyst, as well as in the peritubular connective tissue of cystic seminiferous tubules. In addition, the tunica adventitia of blood vessels within the interstitial space of the cystic seminiferous tubules displayed neurofilament protein, S-100 and protein gene product 9.5 immunoreactive nerve fibres of varying intensity and pattern. Protein gene product 9.5 immunostaining was also observed in the multinucleated giant cells of both the normal and the cystic seminiferous tubules. The cystic portion of the testis appears to have a richer innervation than the normal portion of the same testis. The richer innervation of simple testicular cysts in the ostrich is similar to that observed in the cryptorchid testis of mammals.

The cellular composition of the testicular capsule, seminiferous peritubular tissue, the epithelia as well as periductal muscle cell layers of the excurrent ducts was studied, in sexually mature and active Masked Weaver (Ploceus velatus) birds of the passerine family, Ploceidae. Ultrastructure of the contractile cells in the testicular capsule, peritubular and periductal tissues showed that these cells were smooth muscles of typical morphological characteristics. Variability in the immunohistochemical co-expression of microfilaments and intermediate filaments in the different tissues was evident. Actin and desmin proteins were co-expressed immunohistochemically in the testicular capsule and seminiferous peritubular smooth muscle layer. Actin was singly and very weakly expressed in the rete testis epithelium while cytokeratins and desmin were co-expressed in the epithelium of the excurrent ducts. The periductal muscle layer of all ducts of the epididymis, the ductus deferens as well as the seminal glomus, strongly co-expressed actin and desmin. Vimentin was absent in all cells and tissue types studied. There is clear evidence that the tissues of the male gonad and its excurrent ducts in the Masked Weaver, as has been reported for members of the Galloanserae and Ratitae, contain well-formed contractile tissues whose function would include the transportation of luminal through-flow from the testis into, and through, its excurrent ducts. The microtubule helix in the head and of the mid-piece, of elongating spermatids, as well as of the mature spermatozoa in the various excurrent ducts, including some spermatozoa in the seminal glomus, also co-expressed these three proteins., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The fate of the proximal centriole in passeridan birds is an area of controversy and relative lack of knowledge in avian spermatogenesis and spermatology. This study examines, for the first time, spatiotemporal changes in the centriolar complex in various phases of spermiogenesis in a passerine bird, the Masked weaver (Ploceus velatus). It also describes the configuration of the centriolar complex and the relationship between it and the granular body in both intra- and extra-testicular spermatozoa. It is shown that the proximal centriole is retained and attaches, at its free end, to the granular body of spermatids in every step of spermiogenesis, as well as in mature intra-testicular and post-testicular spermatozoa, including those in the lumen of the seminal glomus. As the centriolar complex, along with its attached granular body, approaches the nucleus in the early spermatid, the proximal centriole articulates with the distal centriole at an acute angle of about 45°, and thereafter, both centrioles, still maintaining this conformation, implant, by means of their articulating proximal ends, at the implantation fossa of the nucleus. In the mature spermatid and spermatozoon, the granular body winds itself helically around the centriolar complex in the neck/midpiece region of the cell, and, thus, becomes the granular helix. The significance of this observation must await future studies, including possible phylogenetic re-evaluation and classification of birds., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The volumetric proportion of the various ducts of the epididymis of the emu and ostrich and the immunohistochemistry of actin microfilaments, as well as cytokeratin, desmin and vimentin intermediate filaments, were studied in the various ducts of the epididymis of the emu and ostrich. The volumetric proportions of various ducts, which are remarkably different from those of members of the Galloanserae monophyly, are as follows: the rete testis, 5.2 +/- 1.4% for the emu and 2.4 +/- 1.8% for the ostrich; efferent ducts, 14.2 +/- 2.3% (emu) and 11.8 +/- 1.8% (ostrich); epididymal duct unit, 25.8 +/- 5.8% (emu) and 26.1 +/- 4.1% (ostrich) and connective tissue and its content, 54.7 +/- 5.8% (emu) and 60.0 +/- 4.9% (ostrich). Unlike in mammals and members of the Galloanserae monophyly, only vimentin was immunohistochemically demonstrated in the rete testis epithelium of the emu, and none of the cytoskeletal protein elements in the ostrich rete testis. The epithelium of the efferent ducts of the emu co-expressed actin, cytokeratin and desmin in the non-ciliated type I cells, and vimentin in the ciliated cell component. The ostrich demonstrated only cytokeratin in this epithelium. The ratite epididymal duct unit is different from that of mammals in lacking actin (only weaky expression in the ostrich), desmin and cytokeratin, and a moderate/strong immunoexpression of vimentin in the basal cells and basal parts of the NC type III cell in the epididymal duct unit. Immunoexpression of the microfilaments and intermediate filaments varied between the two ratite birds, as has been demonstrated previously in birds of the Galloanserae monophyly, and in mammals.

The macroscopic features of the venous drainage of the reproductive system of the male ostrich were studied in six pre-pubertal and three sexually mature and active birds. Each testis was drained by one to four testicular veins. The right testicular veins drained the right testis and epididymis and its appendix to the caudal vena cava and to the right common iliac vein, whereas the left testicular veins drained the left testis and epididymis and its appendix exclusively to the left common iliac vein. A number of variations in the drainage pattern based on the point of entry and number of testicular veins were observed. The cranial aspect of the testis was also linked to the caudal vena cava or common iliac vein via the adrenal veins. The cranial, middle and caudal segments of the ductus deferens (and ureter) were drained by the cranial, middle and caudal ureterodeferential veins respectively, to the caudal testicular veins, the caudal renal veins and pudendal/caudal part of the internal iliac veins. In some specimens, the caudal ureterodeferential veins also drained into the caudal mesenteric vein. The surface of the phallus was drained by tributaries of the pudendal vein. The basic pattern of venous drainage of the reproductive organs of the male ostrich was generally similar to that described for the domestic fowl. However, important differences, including the partial fusion of the caudal renal veins, drainage of the cranial aspect of the testes via the adrenal veins, drainage of the caudal ureterodeferential veins into the caudal mesenteric vein and the presence of veins draining the surface of the phallus, were observed. Although significant, these differences may simply reflect variations in the normal pattern of venous drainage of the reproductive tract of birds which could be verified by studying more specimens and more species.

The presence, location and degree of immunoexpression of various microfilament (MF) and intermediate filament (IF) systems (actin, cytokeratins, desmin, vimentin) were studied in the excurrent ducts of the testis in sexually mature and active galliform (Japanese quail, domestic fowl, turkey) and anseriform (duck) birds. These proteins were variably expressed between the epithelia and periductal tissue (periductal smooth muscle cell layer and interductal connective tissue) types and between species. Variable heterogeneous co-expression of filament systems was also found in the various duct epithelia and periductal tissue types: co-expression of filament systems was the rule rather than the exception. In the duck, neither vimentin nor cytokeratin was present in any of the tissues, whereas actin and desmin (absent in the rete testis) were co-expressed in the efferent ducts and epididymal duct unit (comprising the ductus conjugens, ductus epididymidis and ductus deferens). Actin, desmin and vimentin were generally co-expressed in the rete testis, efferent ducts and epididymal duct unit of the quail, domestic fowl and turkey, with vimentin being more strongly immunoreactive than actin and desmin in the epididymal duct unit, but more weakly immunoexpressed in the efferent ducts. Cytokeratin was present and co-expressed with actin, desmin and vimentin in the rete testis, efferent ducts and epididymal duct unit of the domestic fowl and turkey, but not in the quail and duck. The periductal smooth muscle cell layer and interductal tissue co-expressed actin, desmin and vimentin variably in all birds. Luminal spermatozoa of both the turkey and duck were immunonegative for all protein systems, whereas those of the quail and domestic fowl co-expressed actin, desmin and vimentin moderately or strongly. The tissues of the reproductive tract of male birds thus contain cytoskeletal protein systems that are variably but mostly co-expressed and whose contractile ability appears necessary and sufficient for transportation through the various excurrent ducts of the voluminous testicular fluid and its high sperm content, characteristic features of male avian reproduction.

The epididymal duct unit, comprising the ductus conjugens, ductus epididymidis and ductus deferens, was studied histologically, ultrastructurally and immunohistochemically in five sexually mature and active birds. The main morphological features of the pre-dominant non-ciliated (type III) cell of the epithelial lining of this duct unit include, but are not limited to, a moderately abundant smooth or sparsely granulated endoplasmic reticulum, electron-dense secretory granules and numerous mitochondria in the supranuclear zone of the cytoplasm. A single, large heterogeneous lipid droplet, of unknown function, was characteristically situated immediately proximal to the nucleus. The epithelium is obviously secretory and specifically, of the merocrine, and not apocrine, type of secretion. The epithelium of the epididymal duct unit was only focally and weakly to moderately immunopositive to both actin MF and desmin IF, while the duct unit was immunonegative to cytokeratin and vimentin intermediate filaments. The peritubular muscular layer was moderately to strongly positive to both actin and desmin, and negative to cytokeratins and vimentin.

The testicular capsule and peritubular boundary tissue of the emu and ostrich, as typical representatives of ratite birds, were studied in sexually mature and active birds. The testicular capsule was much thicker (578.1+/-73.4 microm for the free surface of the ostrich testis, and 176.2+/-57.5 microm for the emu) than those of members of the Galloanserae. The cellular composition of both testicular capsule and peritubular tissue was similar generally to that of members of the previously studied Galloanserae and of mammals. The tunica albuginea of the testicular capsule mainly comprised smooth-muscle-like or myoid cells mostly running in one direction and occurring in one main mass. Unlike the Galloanserae, the tunica albuginea contained more collagen fibres than smooth muscle cells, especially in the ostrich. Peritubular tissue was similarly composed of smooth-muscle-like cells distributed in several layers. Actin microfilaments and desmin and vimentin intermediate filaments were variably immunoexpressed in these two tissue types in both birds, with a clear dichotomy in the peritubular tissue. Thus, taken together with studies of some members of the Galloanserae, avian testes clearly contain a morphological mechanism that is represented partly by the smooth muscle cells of the testicular capsule and peritubular tissue for transporting the testicular fluid, which is usually copious in birds, and its cellular content from the testis into the excurrent duct system; this mechanism is similar to that found in mammals.

The macroscopic features of the arterial supply to the reproductive system of the male ostrich was studied in 16 pre-pubertal and eight sexually mature and active birds. The left and right cranial renal arteries arise from the aorta, between the cranial divisions of the kidneys. These vessels supply the cranial divisions of the kidneys, the testes, the epididymides and the cranial segments of the ducti deferentia. Accessory testicular arteries which arise directly from the aorta are present in 45.8% of the specimens. They supply the testes and cranial parts of the ducti deferentia. They are variable in number and origin, and four variants are identified. A cranial ureterodeferential branch originates from the cranial renal artery, supplies the cranial portion of the ductus deferens and ureter, and runs caudally to anastomose with the middle renal artery. The sciatic artery arises laterally from the aorta, just caudal to the acetabulum, and gives rise, ventrally, to a common trunk, the common renal artery, which divides into the middle and caudal renal arteries. The middle renal artery gives rise to the middle ureterodeferential branch which supplies the middle part of the ductus deferens and ureter. A few centimetres caudal to the kidney, the aorta terminates in three branches, namely, the left and right internal iliac arteries and the median caudal artery. The internal iliac artery divides into the lateral caudal artery and the pudendal artery; the latter gives off caudal ureterodeferential branches that supply the caudal segments of the ductus deferens and ureter. In addition, the pudendal artery gives off vessels that supply the cloaca, some of which continue to the base of the phallus, where they form an arterial network. In conclusion, the pattern of the blood supply to the reproductive organs of the male ostrich is, in general, similar to that of the domestic fowl and pigeon, although there are a few highlighted distinctive features.

The testicular capsule was studied histologically, morphometrically, ultrastructurally and immunohistochemically in the Japanese quail, domestic fowl, turkey and duck (all members of the Galloanserae). The testicular capsule was, relative to mammals, thin, being 81.5 +/- 13.7 microm in the quail, 91.7 +/- 6.2 microm in the domestic fowl, 104.5 +/- 29.8 microm in the turkey and 91.8 +/- 18.9 microm in the duck. The orchido-epididymal border (hilus) of the capsule was much thicker than elsewhere in all birds (from 233.7 +/- 50.7 microm in the duck to 550.0 +/- 147.3 microm thick in the turkey). The testicular capsule, other than the tunica serosa and tunica vasculosa, comprised, in the main, smooth muscle-like or myoid cells running mainly in one direction, and disposed in one main mass. Peritubular tissue was similarly composed of smooth muscle-like cells disposed in several layers. Actin and desmin intermediate filaments were immunolocalized in the inner cellular layers of the capsule in the quail, domestic fowl and duck, but uniformly in the turkey. Vimentin intermediate filament immunoreaction in the capsule was moderately and uniformly positive in the testicular capsule of only the quail. Actin and desmin, but not vimentin (except very faintly in the turkey) or cytokeratin, were immunolocalized in the peritubular tissue of all birds. The results therefore establish, or complement, some previous observations that these birds have contractile cells in their testicular capsule and peritubular tissue, whose function probably includes the transport of testicular fluid into the excurrent duct system.

The efferent duct of the ostrich consists of two segments, the proximal efferent duct (PED) and the distal efferent duct (DED) that are continuous, as in some other birds. Both segments of the duct possess an epithelium comprising non-ciliated and ciliated cells in varying proportions between the two segments. The non-ciliated cell (type I) of the PED contains a well-developed, subapical endocytic apparatus of apical tubules and endocytic vacuoles, a solitary, large, heterogeneous lipid droplet, and numerous, oval, dense bodies in the supranuclear region of the cell. Mitochondria tend to concentrate in the basal part of the cell. Intercellular spaces between the non-ciliated cells are enlarged, especially in the basal half of the epithelium. Together, these morphological features confer on the PED an efficient fluid absorption capability. The DED epithelium displays the type II non-ciliated cell whose poorly developed subapical endocytic apparatus as well as the absence of dilated basal intercellular spaces indicate its limited fluid absorptive capacity.

Differences exist in reproductive physiology between African buffalo (Syncerus caffer), cattle (Bos taurus) and water buffalo (Bubalus bubalis). The aim of this study was to histo-morphologically compare the anatomy of non-pregnant and pregnant uteri of buffalo and cattle. Two non-pregnant uteri and placentae of six pregnant African buffalo were used. Early placentome formation (fetal crown rump length (CRL): 2-17.5 cm) in S. caffer and B. taurus was compared. The endometrium of buffalo uteri comprises round to ovoid, dome-shaped and gland-free caruncles. A predominantly simple columnar epithelium of non-ciliated cells covers caruncular tissue, while, additionally, ciliated cells occur in the epithelium of the intercaruncular areas and within the simple columnar or pseudostratified epithelium of the endometrial glands. During early gestation, multiple placentomes develop. Unlike the placentomes in cattle at similar CRL, buffalo placentomes do not develop a caruncular stalk. The sessile, dome-shaped buffalo placentome has simple, slightly conical villi branching less than in cattle, thus indicating different and less complex feto-maternal interdigitation than seen in the latter. A synepitheliochorial interhaemal barrier can be expected in the buffalo placenta, as the occurrence and ultrastructure of trophoblast giant cells resemble those described in cattle.

Carbendazim, a metabolite of benomyl which is widely used as a fungicide, has been found to cause testicular and epididymal damage in laboratory rats, mice and hamsters. No studies of the effects of this chemical on the reproductive organs of birds have been reported previously. This report is that of an investigation on the response of the testis of the Japanese quail to experimental administration of this chemical in sexually mature and active birds. A single dose (400 mg/kg body weight) of carbendazim was administered orally to 20 quails that were sacrificed thereafter at 5 h, 3, 8 and 13 days post-exposure, at five birds spatio-temporally. Five birds acted as control. Testis weights and seminiferous tubular diameter as well as epithelial height decreased significantly from 8 day post-exposure. Epithelial histology was remarkably disrupted, and cessation of spermatogenesis occurred at 13 day post-administration of the chemical. Degenerative changes were uniform in each testis, and not patchy or multi-focal, as previously reported in the rat. The observed histological changes in the testis, due to carbendazim, were capable of causing prolonged infertility in exposed birds.

The response of the epithelial cells of the various ducts of the avian epididymis, whose function is poorly understood, to intraluminal bacteria was evaluated by the injection of an avirulent strain of Salmonella gallinarium into the RT for 24 h. Ultrastructurally, bacteria and invading mononuclear cells were present in the lumina of the RT, proximal efferent ducts (PED) and distal efferent ducts. However, only the non-ciliated (Type I) cells of the PED epithelium ingested bacteria from the lumen. Fragments of bacteria also occurred in several intercellular spaces in the epithelium of the PED. Some mononuclear cells also contained fragments of bacteria. Neither cell death in the various epithelia nor mononuclear infiltration of the periductal tissue occurred. Therefore, in addition to the established function of absorbing most of the testicular fluid entering the epididymis, the Type I cells also appear capable of recognising and removing foreign particulate matter from the epididymal through-flow in the proximal part of the epididymis.

The epithelium of the rete testis of the ostrich is simple cuboidal to columnar in nature, unlike the more flattened epithelium encountered in other non-passerine birds. A solitary cilium projects into the ductal lumen. A unique lateral cell membrane modification, similar, in some respects, to a hemi-desmosome, occurs frequently along the length of this membrane. The lateral cell membrane is complexly folded, and the cell contains an abundance of intermediate filaments, particularly in the vicinity of the nucleus and the internal surface of the basal cell membrane. The Golgi complex and endoplasmic reticulum are moderately developed. Another unique feature of the cell is the presence of a solitary, large, heterogeneous lipid body/droplet situated in the immediate supranuclear region of the cell. Overall, the cell structure conveys the impression of a metabolically active cell, which has the capability of transporting material from the duct lumen to the basal part of the cell, and of protein synthesis, apparently much in excess of what is necessary for cellular maintenance and sustenance.

1. Ultrastructural changes that take place in the spermatids of the turkey were studied. Twelve steps of spermiogenesis are described. 2. Acrosomal formation is similar to that in the domestic fowl and quail. The perforatorium develops as a dense granule in a tubular invagination of the thickened part of the nuclear membrane during the acrosomal phase of spermatid development. 3. Patchy appearances of the circular (CM) and longitudinal (LM) manchette occur concurrently for a very brief period before the complete disappearance of the former and full establishment of the latter, an indication that the CM reorganises to become the LM. 4. A chromatoid body, as found in mammals, is present in the mid-piece of the turkey spermatid, and glycogen aggregations occur in the spermatid cytoplasm, but not in the mature spermatid or spermatozoon. 5. Mitochondrial alignment around the axoneme in the mid-piece takes place only after the dissolution of the LM, unlike in the Japanese quail. 6. The merits and disadvantages of the 4-phase and step-wise systems of spermatid classification and evaluation are discussed.

Simple testicular cysts are rare in mammals and of unknown aetiology, but dietary conditions have been implicated in this phenomenon in poultry. This study characterises the structural features of simple intratesticular cysts in the ostrich. Seven testes from a total of 15 sexually mature ostriches slaughtered during the breeding season were used. The testes contained multifocal, fluid-filled, unilocular cysts which were lined by a simple squamous or low cuboidal epithelium and surrounded by a thick layer of fibrous connective tissue. The cysts developed within seminiferous tubules and displayed morphological features similar to those reported in man and domestic poultry. The testis parenchyma revealed several foci composed of intermingled normal seminiferous tubules and variably sized intratesticular cysts. The atrophic tubules lay within a mass of hyperplastic, fibroblastic intertubular connective tissue in the proximity of large cysts and their formation appeared to result from hydrostatic pressure exerted by cysts. Morphological evidence supports a continuous process of cyst formation in the affected testis and a concomitant progressive loss of atrophic seminiferous tubules. A pathogenetic scenario of cyst formation and the effect of simple cysts on testicular histology has been proposed. Although the course of this phenomenon remains unknown, its impact on the fertility of this economically important bird deserves closer scrutiny.

The structure of the efferent ducts of animals is known to vary from one species to another, it even varies between segments of the ducts in the same species. In the grasscutter or greater cane rat ( Thryonomys swinderianus), there are three segments of the efferent duct, based on their content of non-ciliated or principal cell types. Type I non-ciliated cell is present exclusively in the long proximal part of the duct, and exhibits a well-developed subapical endocytic apparatus as well as numerous oval or pleomorphic dense bodies. The type II non-ciliated cell predominates in the middle part of the duct, displays a poorly developed subapical endocytic apparatus but contains large, numerous vacuoles and dense bodies, all of which fill about two-thirds of the cell height. The type III non-ciliated cell, found in the epithelium of the terminal part of the duct, is poorly endowed with a subapical endocytic apparatus and contains no conspicuous endocytic vesicles or vacuoles. Only a few, small, dense bodies are present, if at all. The efferent duct of the cane rat is thus similar to that of man, the bull, goat and dog, in containing three varieties or types of non-ciliated cells. This report is the first to describe multiple non-ciliated cells in the epithelium of the efferent ducts of a rodent and, indeed, of a mammal smaller than the dog.

The changes that take place in the efferent ducts during the major phases of the reproductive cycle of birds were studied morphologically using standard histological, morphometric, and ultrastructural methods in prepuberal, sexually mature and sexually active, and sexually mature but sexually inactive domestic fowl (Gallus domesticus), drake (Anas platyrhynchos), and guinea fowl (Numida meleagris). Profound structural and dimensional changes occurred in both segments (proximal and distal) of the efferent ducts and, in particular, in the nonciliated (Type I) cell of the proximal duct of sexually mature but inactive birds. The subapical tubulovacuolar system was markedly atrophic in nonciliated (Types I and II) cells and the numerous round dense globules of Type I cells that normally occurred in sexually active birds were replaced by fewer and more pleomorphic bodies containing lipofuscin granules in sexually resting birds. Lipid droplets, few and extremely large in inactive drakes but numerous and smaller in size in guinea fowls and domestic fowls, occurred in the Type I cell at both infra- and supranuclear levels of resting but not in prepuberal or sexually active birds. Ciliated cells in both segments of the ducts exhibited fewer and less profound phase-dependent changes ultrastructurally. Generally, the Type I cells of the proximal efferent duct appeared to be more sensitive to androgen deprivation than the Type II cell of the distal efferent duct or ciliated cells in both ducts. These morphologically phase-dependent features of the efferent ducts of birds may be used, together with or independent of testicular changes, in the determination of the status of the testis and epididymis of a male bird with regard to the reproductive cycle, especially in seasonally breeding species., (Copyright 2002 Wiley-Liss, Inc.)

As part of a series of studies designed to evaluate usual and potential functions of epithelial cells in the avian epididymis under varying circumstances, adult, sexually active male Japanese quails were vasectomised for 3, 6 and 8 weeks. The testes, epididymides and deferent ducts were studied histologically and ultrastructurally. Testicular lesions were minimal, but epididymal response was profound. Non-ciliated (type I) cells of the proximal efferent duct (PED) actively phagocytized spermatozoa and their fragments. An especially noteworthy and apparently novel observation was the proliferation of the lining epithelium in parts of the PED to form additional adluminal sheets of spermiophagic epithelium which did not lie directly on basal laminae. These sheets were composed exclusively of non-ciliated cells. Spermatozoal dissolution and macrophage activity were also important mechanisms for the disposal of superfluous and incarcerated spermatozoa in the epididymis of this bird., (Copyright 2002 Published by Elsevier Science Ltd.)

The epididymis of two species of domestic birds, the domestic fowl (Gallus domesticus), duck (Anas platyrhynchos), and of domestic and feral guinea-fowl (Numida meleagris) was studied during the three main phases of the reproductive cycle (prepuberal, sexually mature and active, and sexually mature but inactive or resting) with a view to identifying major histological and ultrastructural changes associated with and distinctive for each phase. Rete testis cells accumulated numerous variably sized lipid droplets in all birds, as well as large heterogeneous and lipofuscin-containing dense bodies in the guinea-fowl, during the resting but not in the other phases. The principal or Type III cells of the connecting and epididymal ducts exhibited profound structural changes, including, but not limited to, rarefied cytoplasm, inconspicuous and general loss of sparsely granular endoplasmic reticulum, loss of secretory vesicles in the drake, and an enhanced and conspicuous presence of lipid droplets in the guinea-fowl. The rete cells appeared to be less sensitive than the Type III cells to a reduced level or absence of lumenal androgens. These phase-dependent changes may help to prevent or minimize discrepancies in the interpretation of the normal structure of the epididymis in birds during the sexually active phase, as distinct from the other two phases and their intermediate phases.

Indigenous Sabi rams of Southern Africa were experimentally infected with Trypanosoma congolense for 8 and 16 weeks. Testes weights (g) were significantly (P<0.05) lower in the infected (249.7+/-26.4) compared to the control (372.63+/-19.4) animals. Histopathological and ultrastructural changes included seminiferous tubular atrophy and mononuclear infiltration in the testis, and lesions in the epithelium of the corpus epididymidis (middle segment) as well as spermatozoa in the cauda epididymidis. The gonadal lesions may have the capability to impair fertility in Sabi rams infected with Trypanosoma congolense.

The various ducts of the epididymides of four gallinaceous birds, the turkey (Meleagris gallopavo), domestic fowl (Gallus gallus), guinea-fowl (Numida meleagris) and Japanese quail (Coturnix coturnix japonica) were studied at the scanning and transmission electron microscopy levels. The tissues were fixed either by immersion or vascular perfusion, for comparative purposes. Each duct system, save for a few details, presented similar morphological features in all species. The epithelial surface of the rete testis was regular and each cell bore a single cilium, as well as numerous, or in some parts, very few, short, regular microvilli. Each of the Types I and II non-ciliated cells of the proximal and efferent ducts displayed abundant, moderately long and regular microvilli, and a solitary cilium. The ciliated cells exhibited tufts of cilia. The Type III non-ciliated cell of the connecting and epididymal ducts exhibited a solitary cilium, and numerous microvilli which were intermediate in length between those of the rete testis and those of the efferent ducts. Vascular perfusion of the avian epididymal tissue was the superior method of fixation because it minimised the developments of fixation artefacts. Apocrine secretion did not appear to occur in the epididymis of these birds as the apical blebs of Types I, II and III cells, which have previously been reported, only manifest in this study in inadequately fixed tissues, and were therefore viewed as being artefacts. The present findings suggest that the current terminology, as applied to the avian epididymis, be retained.

The structure of the ductus epididymidis, and its capacity to take up luminal particulate material (India ink) were studied, using scanning and transmission electron microscopy, in the domestic fowl, turkey and Japanese quail (Order Galliformes) and drake (Order Anseriformes). The epithelium appeared contractile and stable structurally, especially in the drake in which, among other cytoskeletal features, abundant perinuclear intermediate filaments radiated peripherally in the principal, nonciliated (Type III) cell. The Golgi complex was well developed, except in the turkey in which it was extremely inconspicuous. Sparsely granulated profiles were the main component of the elaborate endoplasmic reticulum, which was best developed in the drake and least in the turkey. The cell in the turkey was uniquely laden with large aggregates of lipid droplets. Generally, characteristics of a cell active in the synthesis and merocrine secretion of proteinaceous material were evident. There was no evidence of apocrine-like secretion in well-fixed tissues. The cell lacked the capacity to take up luminal India ink particles, and displayed very poorly developed endocytic structural features, which are also probably only selectively and minimally absorptive. Ultrastructural features varied between and within the orders studied, necessitating further studies.

In a study of the absorptive activities of the excurrent ducts of the testis of birds, six adult cocks of the Ovambo breed of domestic fowl were deeply anaesthetised and intravascularly perfused with buffered 3 per cent glutaraldehyde. Epididymal tissue was prepared conventionally for electron microscopy. In favourable sections in three of these birds avid ingestion of spermatozoa was revealed in the non-ciliated (Type I) cells of the epithelial lining of the initial part of the proximal efferent duct, at and a little beyond its junction with the rete testis. No obvious defects were detected in luminal spermatozoa. The testicular excurrent ducts were neither distended nor obstructed, and mononuclear cell mobilisation was absent in both the ductal and periductal tissue. The significance of this observation with regard to the specific segment of the efferent duct system involved and how certain spermatozoa are identified for elimination from the duct lumen, must await precise studies., (Copyright 2000 Harcourt Publishers Ltd.)

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.

The interstitial tissue of the testis was studied in gonadally active and gonadally inactive domestic fowl (Gallus gallus domesticus), guinea-fowl (Numida meleagris), duck (Anas platyrhynchos) and Japanese quail (Coturnix coturnix japonica). Gonadal inactivity in the fowl was induced by a single subcutaneous injection of 50 mg oestradiol benzoate. The structure of this tissue was similar in all the birds studied. Lymphatic vessels were mostly thin and meandered between the peritubular tissue peripherally and the centrally located blood vessels, Leydig cells and macrophages. The basal lamina rested on a closely associated homogeneous microfibrillar layer free of collagen fibres. The myofibroblast layer was several cells thick, and quite compact. The basal lamina of gonadally resting birds was highly irregular, relatively electron-dense, contained electron-lucent globules, and sent numerous finger-like processes or plicae into the seminiferous epithelium, particularly into the Sertoli cells. The Leydig cells were few but typical in structure. In gonadally inactive birds they accumulated lipid droplets, dense heterogeneous bodies, probably lysosomes, and appeared to degenerate. The avian testicular interstitium is similar to that of the human and cat in possessing a multi-layered myofibroblast component, and to that of the rodent in possessing a small number of Leydig cells, as well as in the location of the lymphatic vessels. Thus the bird combines characteristics of the interstitium found variably in mammals.

Parameters were obtained from the reproductive organs of ethanol-fed, gossypol-treated Sprague Dawley rats. The experimental animals were fed either on a normal (15.17%) or low protein (8.00%) diet. Measurements included reproductive organ weights, seminal characteristics, serum concentration of testosterone and histological, stereological and histomorphometric evaluation of the testis. The testis size, length and diameter of the seminiferous tubule had the least values in the protein-malnourished, gossypol-treated rats (3.01 +/- 0.26 g, 0.56 +/- 0.03 m, 281.34 +/- 11.30 microns), in comparison to corresponding animals which had received ethanol simultaneously with gossypol (3.40 +/- 0.25, 0.71 +/- 0.06 m, 314.42 +/- 11.61 microns). As gossypol and ethanol are both associated with reduced reproductive capacity, this unexpected but interesting finding lends support to the hypothesis that either a normal dietary protein or ethanol consumption may modify the action of gossypol on body tissues, including the testis. This effect, presumably mediated through changes caused to the bioavailability of gossypol, modifies its antifertility activity. The present observation further highlights the need to consider the concurrent administration of other drugs, such as alcohol, and the nutritional status in the evaluation of gossypol for various potential uses.

The testes of adult male Wistar rats that were variously protein malnourished, gossypol treated, and/or trypanosome infected (Trypanosoma brucei) were evaluated ultrastructurally. The findings included several necrotic germ cells in the seminiferous epithelium and focal degeneration of Sertoli cells. Leydig cells showed a remarkable paucity of smooth endoplasmic reticulum, and aggregation of mitochondria. These observations were prevalent in those animals that were simultaneously protein-malnourished/gossypol treated/ trypanosome infected, and were either absent or markedly reduced in the other groups. These structural alterations are probably related to increased gossypol availability to the testis and/or represent the additive effects of gossypol and trypanosomes. Such tissue changes could compromise spermatogenesis in affected animals, and suggest that trypanosome infections may exacerbate testicular lesions in the gossypol-treated rat.

The epithelium of the vesicular gland of the African giant rat was studied by the light and electron microscopes. The gland is compound tubuloalveolar, lined by a columnar epithelium, the apical surface of which is covered by a few short microvilli. The epithelial cells are the principal and basal cells. The principal cells have an abundance of rough surfaced endoplasmic reticulum (rER), which are often arranged in whorls. Basal cells are few, being wedged between adjacent principal cells in the basal region. They are fairly electron lucent and contain fewer organelles. Intraepithelial lymphocytes, which also occur mostly in the basal region are also present in the epithelium. The presence of these latter cells is noteworthy in that they are not a known feature of the epithelium of the vesicular gland of other species.

Some aspects of the toxicity of gossypol were investigated in ethanol-fed, gossypol-treated Sprague-Dawley rats, with and without protein malnutrition. Serum creatine phosphokinase activity was depressed in all gossypol-treated rats and gossypol caused greater decreases than ethanol in those animals that were fed the normal diet. Even though the adrenal glands were enlarged in all gossypol-treated rats, this was accompanied by depressed serum cortisol levels. Histopathological findings, including vacuolar degeneration of hepatocytes and dilation of the sinusoids in the liver, atrophy and leucocytic infiltration of cardiac myofibres, vacuolation of the renal glomerulus and cellular depletion of the spleen and thymus, were most severe in the protein-malnourished, gossypol-treated rats; these were generally absent in the other groups. These findings indicated that either adequate levels of protein in the diet and/or ethanol consumption have the effect of reducing gossypol toxicity, probably by reducing the amount of gossypol available for tissue exposure through enhanced metabolism and/or clearance from the body. However, an evaluation of the metabolic pathways involved in gossypol metabolism in ethanol-fed individuals might provide a better understanding of these observations.

The toxicity of gossypol, a compound occurring naturally in the cotton plant, was investigated in Trypanosoma brucei-infected, gossypol-treated rats, with and without protein malnutrition. The liver, heart, lungs, spleen and adrenal glands were enlarged in all gossypol-treated rats. Gossypol treatment or trypanosome infection, either alone or together, invariably caused significant reductions in the serum activity of creatine phosphokinase and amylase and in the serum concentration of cortisol. The serum biochemical changes, together with histopathological findings in various organs, indicated that the toxicity of gossypol and pathology of trypanosome infection, either alone or in concert, could be exacerbated by protein malnutrition. This finding suggests that the previously reported antiparasitic properties of gossypol may be of little ultimate benefit due to these serious side effects. The spleen in the protein-malnourished, trypanosome-infected and gossypol-treated animals exhibited only a slight decrease in the number of lymphatic nodules, but a marked cellular depletion, especially of cortical tissue, was observed in the thymus. These observations would seem to justify further study of the immune status of trypanosome-infected, gossypol-treated animals.

Reproductive parameters were investigated in gossypol-treated male rats that had been infected experimentally with Trypanosoma brucei and then place on one of two diets of differing (low and normal) protein content. The endpoints assessed were reproductive organ weights, semen epididymal sperm counts, serum testosterone and histological, stereological and histomorphometric evaluation of the testis and accessory reproductive organs. Most of the parameters studied were lowest in the protein-malnourished, gossypol-treated, trypanosome-infected animals, when compared to those obtained from the corresponding animals that were only gossypol-treated or trypanosome-infected. Mean testicular size and the diameter and the total length of seminiferous tubules were especially reduced, indicating that the overall volume of the seminiferous epithelium in these animals was smaller. These findings suggest that reproductive capacity could be impaired in protein-malnourished, trypanosome-infected animals fed on gossypol-containing products, even when there are no obvious clinical signs of disease. This could translate into increased production costs in a farm enterprise. Screening for haemoparasitism would also seem to be worthwhile prior to evaluation and/or use of gossypol (or perhaps other potential contraceptives) in human subjects, especially in communities that are confronted with severe food shortages.

Serum sodium (Na), potassium (K), chloride (Cl), phosphate (Po), and calcium (Ca) were assayed in adult male Wistar rats fed a low protein diet (LP) and a normal protein diet (control, NP) and administered gossypol (GP) and chloroquine (CQ) alone and in combination (GPCQ), for 4 and 8 weeks. Gossypol was administered at 20 mg/kg body wt/rat/day in Goya oil and chloroquine at 5 mg/kg body wt/rat for 3 days/week in distilled water. The data, analyzed by two-way ANOVA, showed that the interaction had significant effects on the serum levels of Na, K and Po (P < 0.001) but not on chloride and calcium (P > 0.05). Serum levels of sodium, potassium and phosphate in LP rats were significantly lower than in NP rats. In LP and NP rats, the levels of Na, K, and Po at 8 weeks were significantly higher than at 4 weeks; and GPCQ treatment produced significantly lower sodium levels in LP and NP rats, lower phosphate in LP rats and higher phosphate in NP-fed rats than GP or CQ administered alone. Potassium level in GPCQ was between levels in Gp- and CQ-treated rats. The administration of gossypol and chloroquine together may not adversely affect serum electrolytes in protein-malnutrition state any more than gossypol or chloroquine administered alone.

Gossypol and chloroquine were administered in goya oil and distilled water, respectively, to low protein-fed (LP) and normal protein-fed (NP, control) adult male albino Wistar rats for 4 and 8 weeks' duration. Gossypol (GP) and chloroquine (CQ) were administered separately and in combination (GPCQ). At the end of the treatment periods, the rats were anesthetized and blood drawn by cardiac puncture, centrifuged, and sera obtained for assay of total cholesterol and triglycerides levels. The data obtained were analyzed by two-way ANOVA. The results showed that the interaction of GP and CQ in LP and NP-fed rats produced significant effects (P<0.001) on the serum levels of cholesterol and triglycerides. Serum cholesterol level was lower at 8 than at 4 weeks in LP-fed rats but higher at 8 than at 4 weeks in LP fed rats. The administration of GP, CQ, and GPCQ resulted in lower serum cholesterol and triglycerides at 8 than at 4 weeks in LP fed rats but higher serum cholesterol at 4 than at 8 weeks in NP-fed rats. In LP and NP-fed rats, the levels of serum cholesterol and triglycerides were generally lower in GCPQ-treated than GP- or CQ-treated rats. The implications of the findings are discussed.

This study was designed to evaluate the interaction between protein malnutrition, gossypol treatment and blood parasitosis (Trypanosoma brucei) in the Wistar rat. Haematological and serum biochemical changes were evaluated in the rats, which were placed on two planes of nutrition--low protein (LP) and normal protein (NP)--and either treated with gossypol or infected with Trypanosoma brucei, or both. Higher parasitaemia occurred in gossypol-treated NP rats than in the corresponding LP group. Gossypol treatment and trypanosomal infection, either alone or in concert, caused an anaemia that was both macrocytic and hypochromic. Both treatments together also caused increases in serum alkaline phosphatase and alanine aminotransferase activities, which were accompanied by depressed serum albumin concentrations, suggestive of hepatic dysfunction in affected rats. These results suggest that, with adequate protein intake, the growth and infectivity of trypanosomes is not inhibited by gossypol but that protein malnutrition has a beneficial effect of reduced parasitaemia. Unfortunately, this beneficial effect is counteracted by gossypol enhancement of hepatic dysfunction caused by trypanosomes.

The effects of protein malnutrition on haematological and serum biochemical values were evaluated in gossypol-treated rats which were simultaneously fed with ethanol. Gossypol caused anaemia, leucopenia and thrombocytopenia in malnourished animals, suggesting a depression of bone marrow activity. Gossypol also caused a significant elevation of serum alkaline phosphatase and alanine aminotransferase activities and increases in the concentrations of Mg++ and Ca++ with reduced albumin, regardless of the nutritional status. These changes were more severe with malnutrition. Ethanol alone caused a thrombocytopenia but no other significant haematological changes. However, it appeared to cause derangement of lipid and protein metabolism as reflected in serum cholesterol and urea. The toxic effects seen in gossypol-treated rats were significantly reduced in animals simultaneously given ethanol. As the livers of gossypol-treated rats were significantly heavier than in these animals, it seems possible that ethanol consumption enhances the ability of the liver to metabolize gossypol, thereby reducing its accumulation and consequently its toxicity. However, further studies are needed to determine the mechanisms responsible.