A case of histologically diagnosed tick infestation on the scalp of a Korean child

A scalp mass surgically excised from a 4-year-old Korean boy was identified as a tick through histological observations. In sections of the mass, characteristic features of a tick, including its gross contour, cuticular structures, well developed musculature and salivary glands, and the capitulum, were discovered. In particular, the capitulum is anteriorly protruded, which strongly suggests that the specimen be a hard tick of family Ixodidae. However, the present histological features were not enough to determine the genus and species of the tick, because information on sectional morphologies of different tick species is unavailable. This is a rare case of tick infestation on the scalp diagnosed in histological sections.     

Gonochorism and seasonal variations in the gonads of the labrid Symphodus (Crenilabrus) ocellatus (Forsskål)

A histological study of gonadal organization of 354 Symphodus ocellatus caught in the Bay of Naples from June 1985 to December 1986 demonstrated the absence of sexual inversion in this species. Four phases of gonadal development, related to the seasonal cycle, are described. During the reproductive period the ovaries showed the asynchrony in oocyte development that is typical of serial spawners. The testes of three types of males present in this species (territorial, satellite and sneaker males) were examined: the only difference found was in the gonadal growth pattern during the reproductive period.     

Early development of the chondrocranium in Salmo letnica(Karaman, 1924)(Teleostei: Salmonidae)

The ontogenetic development of the chondrocranium of Ohrid trout Salmo letnica was studied from hatching until 92 days post‐hatching (dph). Most of the samples were in toto trypsin cleared and stained, some specimens were used for serial histological sectioning. The serial histological sections of fish specimens at the age of 92 dph were used for a graphical reconstruction of the cartilaginous neurocranium. A chronological evaluation of the formation of the cartilaginous skull in the early development of S. letnica was performed. In order to investigate to what degree the ontogeny of the Ohrid trout is unique, the results were compared with data of the development of other salmonids, as well as some non‐salmonid teleosts. The development of the cartilaginous structures of the Ohrid trout was found to be similar to that of other salmonids. Most of the cartilage structures of the neurocranium and the viscerocranium are present at the moment of hatching of this species. A fully developed chondrocranium was observed at the age of 92 dph, when the first signs of cartilage resorption could also be observed.     

Born among the ice: first morphological observations on two developmental stages of the Antarctic silverfish Pleuragramma antarcticum, a key species of the Southern Ocean

The Antarctic silverfish Pleuragramma antarcticum is a keystone species in the Southern Ocean ecosystem, providing one of the major links between lower and higher trophic levels. Despite the importance of this species, surprisingly little is known of its early development. The first spawning area for the silverfish has been recently identified in the near-shore of Terra Nova Bay (Ross Sea). Evidence indicates that spawning and embryo development occurs in the cryopelagic environment, below the seasonal pack-ice. In order to contribute to the knowledge of the life cycle of this very important Antarctic species, we carried out the first histological characterization on pre-hatching embryos and newly hatched larvae. Embryonated eggs and larvae of P. antarcticum were collected between late October and November 2005 at TNB through holes drilled into the sea ice. Embryonic stage just before hatching and the first post-hatching stage were the most abundant within our samples and thus were analysed using both macroscopic and histological approaches. Early life stages of the Antarctic silverfish revealed interesting features: the sensory system, foraging apparatus and heart appeared well developed, whereas the liver and gills were underdeveloped. Morphological details of the organogenesis were performed, providing the first substantial information on the development of P. antarcticum and representing a further steps towards the knowledge of the life cycle of this important Antarctic key species. An erratum to this article can be found at     

Gonad morphology,oocyte development and spawning cycle of the calanoid copepod <Emphasis Type="Italic">Acartia clausi</Emphasis>

Information on gonad morphology and its relation to basic reproductive parameters such as clutch size and spawning frequency is lacking for Acartia clausi, a dominant calanoid copepod of the North Sea. To fill this gap, females of this species were sampled at Helgoland Roads from mid March to late May 2001. Gonad structure and oogenesis were studied using a combination of histology and whole-body-analysis. In addition, clutch size and spawning frequency were determined in incubation experiments, during which individual females were monitored at short intervals for 8 and 12 h, respectively. The histological analysis revealed that the ovary of A. clausi is w-shaped with two distinct tips pointing posteriorly. It is slightly different from that of other Acartia species and of other copepod taxa. From the ovary, two anterior diverticula extend into the head region, and two posterior diverticula extend to the genital opening in the abdomen. Developing oocytes change in shape and size, and in the appearance of the nucleus and the ooplasm. Based on these morphological characteristics, different oocyte development stages (OS) were identified. Mitotically dividing oogonia and young oocytes (OS 0) were restricted to the ovary, whereas vitellogenic oocytes (OS 1–4) were present in the diverticula. The development stage of the oocytes increased with distance to the ovary in both, anterior and posterior diverticula. Most advanced oocytes were situated ventrally, and their number varied between 1 and 18, at a median of 4. All oocyte development stages co-occur indicating that oogenesis in A. clausi is a continuous process. These morphological features reflect the reproductive traits of this species. In accordance with the low numbers of mature oocytes in the gonads, females usually produced small clutches of one to five eggs. Clutches were released throughout the entire observation period at intervals of 90 min (median) resulting in mean egg production rates of 18–28 eggs female⁻¹ day⁻¹.     

Juvenile bisexuality in the red sea bream,Pagrus major

The histology of the gonad of the red sea bream,Pagrus major, was examined in order to study the early gonadal development, sexual maturation and sex ratio in a natural population. A total of 1,117 fish between the ages of 4 months and 8 years were examined. Gonads of 4-month-old fish were either sexually undifferentiated with a central cavity, or ovarian in form. Gonads of 12- and 18-month-old fish were ovaries or bisexual gonads, while those of 2-year-old fish were ovaries, bisexual gonads or testes. Fish aged between 3 and 8 years had ovaries or testes, except for a few bisexual gonads found in 3- and 4-year-old fish. The chronological appearance of females, hermaphrodites and males in that order, and histological evidence, suggested that the testis originates from the ovary via a bisexual gonad in the juvenile stage. The sex ratio of females to males at the age of 2 years and over was about 1:1, suggesting that hermaphroditic red sea bream appear in about 50% of the juvenile population. The sexual pattern in this species, therefore, is concluded to be gonochorism with a bisexual juvenile stage.     

Histological development of the digestive system of the Amazonian pimelodid catfish Pseudoplatystoma punctifer

The organogenesis of the digestive system was described in the Amazonian pimelodid catfish species Pseudoplatystoma punctifer from hatching (3.5 mm total length, TL) to 41 days post-fertilization (dpf) (58.1 mm TL) reared at 28°C. Newly hatched larvae showed a simple digestive tract, which appeared as a straight undifferentiated and unfolded tube lined by a single layer of columnar epithelial cells (future enterocytes). During the endogenous feeding period, comprised between 20 and 96 h post-fertilization (3.5 to 6.1 mm TL), the larval digestive system experienced a fast transformation with the almost complete development and differentiation of most of digestive organs (buccopahrynx, oesophagus, intestine, liver and exocrine pancreas). Yolk reserves were not completely depleted at the onset of exogenous feeding (4 dpf, 6.1 mm TL), and a period of mixed nutrition was observed up to 6 to 7 dpf (6.8 to 7.3 mm TL) when yolk was definitively exhausted. The stomach was the organ that latest achieved its complete differentiation, characterized by the development of abundant gastric glands in the fundic stomach between 10 and 15 dpf (10.9 to 15.8 mm TL) and the formation of the pyloric sphincter at the junction of the pyloric stomach and the anterior intestine at 15 dpf (15.8 mm TL). The above-mentioned morphological and histological features observed suggested the achievement of a digestive system characteristic of P. punctifer juveniles and adults. The ontogeny of the digestive system in P. punctifer followed the same general pattern as in most Siluriform species so far, although some species-specific differences in the timing of differentiation of several digestive structures were noted, which might be related to different reproductive guilds, egg and larval size or even different larval rearing practices. According to present findings on the histological development of the digestive system in P. punctifer, some recommendations regarding the rearing practices of this species are also provided in order to improve the actual larval rearing techniques of this fast-growing Neotropical catfish species.     

Morpho-histological and ultrastructural study on direct somatic embryogenesis of Capsicum chinense Jacq. in liquid medium

Somatic embryo-like structures were produced from the hypocotyls of aseptic plants of Capsicum chinense. Different concentrations of 2,4-dichlorophenoxyacetic acid (0, 4.5, 9.05 μM), several exposure times of the explant to this auxin (15, 30, 45, 60 days) and the development of somatic embryos cultured in a solid and/or liquid medium were evaluated. As a result, a novel system of regeneration via direct somatic embryogenesis in liquid medium was established, with an efficiency of 1.77 × 10⁴ somatic embryos per liter of medium. Critical stages of embryogenesis, including cellular acquisition of morphogenetic competence, suspensor formation, and development and maturation of somatic embryos, were identified by histological analysis and scanning electron microscopy. Our results show a promising new outlook on the in vitro regeneration of this species. Contrary to what has been reported to date for the Capsicum genus, it is a species of plants with higher embryogenic potential in vitro.     

Histomorphological study on embryogenesis of the honeybee Apis cerana

As important pollination species, honeybees play substantial impacts on the balance of global ecosystem, including two best-known honeybees Apis mellifera and Apis cerana. Embryogenesis is a fundamental stage of honeybee development and plays important roles in supporting the whole-life developmental process. However, few studies were reported on honeybee embryonic morphology using egg section, possibly due to the fragility of honeybee eggs and the difficulty of making embryonic sections. In this study, we reported a simply equipped method of frozen sectioning and PI (propidium iodide) staining to show the inner structure and cell distribution of A. cerena embryos at the different embryonic developmental stages. We found that the stages of A. cerena embryogenesis could also be typically classified into ten developmental stages, which are similar with the sister honeybee species, A. mellifera. To be noted, besides the cell distribution in the whole egg, we clearly observed the migration route of embryonic cells during the early embryonic development in A. cerena. This study provides a new insight into the whole process of honeybee embryogenesis from the perspective of egg sectioning, a histological basis for genetic manipulation using A. cerena eggs, and a reference method for egg sectioning for other insect species.     

Growth and development of Haemonchus contortus in jirds, Meriones unguiculatus.

Growth and development of Haemonchus contortus were examined in jirds and were compared to these processes in lambs. Number, sex, size, and stage of development were determined for worms recovered at necropsy at various times postinoculation (PI) from immunosuppressed jirds inoculated with approximately 1,000 exsheathed infective larvae (L3) of H. contortus. In addition, gastric tissue samples from jirds were examined histologically. Parallel studies were done in lambs inoculated with approximately 7,500 L3. Typically, 5-30% of the inoculum established and survived in jirds at reasonably stable numbers to day 14 PI. By day 21 PI, worm numbers in jirds decreased dramatically. Although the parasite was similar in size and development on day 4 PI in jirds and lambs, from day 7 PI on, worms were significantly smaller and less developed in jirds. On histological examination, the parasite was found only in the glandular portion of the stomach of jirds (anatomically similar to its predilection site in the abomasum of lambs), and histological changes were consistent for both host species. Although growth and development of H. contortus are slower and incomplete in jirds, the parasite establishes, grows, and develops (at anatomically comparable sites in both hosts) in this model. Thus, the model appears to provide a useful laboratory host to study H. contortus.     

Study of Trypanosoma cruzi in wild reservoirs in the ecological reserve of El Zapotal, Chiapas, Mexico

Reservoirs of T. cruzi were studied in the ecological reserve "El Zapotal", in the Tuxtla Gutiérrez municipality located in the state of Chiapas. The parasite transmission cycle exists in that area. A total of 73 specimens of 8 different mammal species were trapped alive. Xenodiagnosis tests showed that 11% of them were positive for T. cruzi. Blood concentration exam, blood culture, serology (CIEP) and histological exam were carried out in 65 specimens from the total captured. T. cruzi positive results in these tests were 1.5, 3.6, and 6.2% respectively. None of the 73 specimens were positive in the histological exams. Amastigote nests were found in cardiac tissue of mice, which were inoculated with triatomine bugs dejections collected from those specimens used from xenodiagnosis testing or by the inoculation of parasites grown in axenic blood cultures. These results indicated that there is a sylvan cycle which is maintained by mammal species, which are plundering this geographic area.     

Follicular development in cryopreserved Common Wombat ovarian tissue xenografted to Nude rats

The Northern Hairy-nosed Wombat (Lasiorhinus krefftii) is a highly endangered marsupial species and every possible option for sustaining the species needs to be explored. One important approach may be the development of assisted reproductive technologies in the non-endangered Common Wombat (Vombatus ursinus) and Southern Hairy-nosed Wombat (Lasiorhinus latifrons) for application in breeding the Northern Hairy-nosed Wombat.In this study, it was examined whether cryopreserved Wombat ovarian tissue would develop following xenografting to immunologically deficient rats. Ovarian tissue was collected from Common Wombats (n = 3) and cryopreserved as small cortical pieces. After thawing the cortical pieces were grafted underneath the kidney capsule of Nude rats (n = 16). The grafts were recovered at 2, 4, and 10 weeks after transplantation and their gross and histological appearance investigated. Two weeks after grafting (n = 2), the tissue was revascularized and healthy primordial follicles were present. At week 4 (n = 2), some follicular development was present. At week 10, six rats received human chorionic gonadotrophin (hCG) to trigger follicle and oocyte maturation while another six rats were not given any treatment. The administration of hCG did not induce preovulatory follicles and oocyte maturation although type 5 follicles were present in ovarian tissue collected 10 weeks posttransplantation in both treated and untreated groups. This study demonstrates for the first time that Wombat ovarian tissue can survive and function when grafted into immunocompromized rats and that Wombat ovarian follicles can be recruited to growth and development in an ovarian xenograft. This model system has the potential to produce mature oocytes from endangered species for use in assisted reproductive technologies such as in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), and mature oocytes from non-endangered species for nuclear transfer which may be necessary for the preservation of critically endangered species.     

Synthetic peptides derived from the Listeria monocytogenes p60 protein as antigens for the generation of polyclonal antibodies specific for secreted cell-free L. monocytogenes p60 proteins.

All species of the genus Listeria secrete a major extracellular protein called p60. A comparison of the deduced amino acid sequences of all listerial p60 proteins previously indicated there were only a few regions which were unique to the pathogenic, food-borne species Listeria monocytogenes. Two of these p60 regions were chosen for the development of antibodies specific for the facultative intracellular species L. monocytogenes. Initially, these regions were characterized via epitope mapping, and this led to the development of two different synthetic peptides. Rabbits immunized with these synthetic peptides generated polyclonal antibodies that were then used in Western blot (immunoblot) analyses. Antiserum against peptide A (PepA) recognized the p60 protein in the supernatants collected from most L. monocytogenes serotypes except for several strains belonging to serotypes 4a and 4c. No p60-related protein was detected in the supernatants from other Listeria species with this anti-PepA antiserum. Antibodies raised against peptide D (PepD) reacted with p60 from all L. monocytogenes serotypes, including all 4a and 4c strains that were tested, and also showed no cross-reactivity with supernatant proteins from other Listeria species. Both antisera also detected p60 in supernatants of a large number of environmental isolates of L. monocytogenes. Besides Western blot analyses, these antisera to PepA and PepD reacted with secreted p60 in an enzyme-linked immunosorbent assay, indicating recognition of the native antigen in addition to the denatured form. These data suggest that synthetic peptides derived from the variable region of the L. monocytogenes p60 protein may be useful for the development of an immunological diagnostic assay.     

A comparative study of the urinary bladder and the intramural portion of the ureter.

The bladder belonging to eleven mammalian species were investigated, and as a result the following observations were made: (1) a submucous ureter was found in the case of most of the species examined; (2) histological investigation revealed three types of intramural ureters, and (3) downward extension of ureteric musculature, separate from bladder musculature, to the seminal colliculus in the male, or to the lower end of the urethra in the female, was found. A suggestion is presented to explain the manner by which the ureteric openings change their relations to those of the mesonephric ducts development. It seems that 'after absorption of the common segment of the Wolffian duct, breaking down of the ventral wall of ureter' is the most likely process.     

Correlation between ultrastructural differentiation of bacteroids and nitrogen fixation in alfalfa nodules.

Bacteroid differentiation was examined in developing and mature alfalfa nodules elicited by wild-type or Fix- mutant strains of Rhizobium meliloti. Ultrastructural studies of wild-type nodules distinguished five steps in bacteroid differentiation (types 1 to 5), each being restricted to a well-defined histological region of the nodule. Correlative studies between nodule development, bacteroid differentiation, and acetylene reduction showed that nitrogenase activity was always associated with the differentiation of the distal zone III of the nodule. In this region, the invaded cells were filled with heterogeneous type 4 bacteroids, the cytoplasm of which displayed an alternation of areas enriched with ribosomes or with DNA fibrils. Cytological studies of complementary halves of transversally sectioned mature nodules confirmed that type 4 bacteroids were always observed in the half of the nodule expressing nitrogenase activity, while the presence of type 5 bacteroids could never be correlated with acetylene reduction. Bacteria with a transposon Tn5 insertion in pSym fix genes elicited the development of Fix- nodules in which bacteroids could not develop into the last two ultrastructural types. The use of mutant strains deleted of DNA fragments bearing functional reiterated pSym fix genes and complemented with recombinant plasmids, each carrying one of these fragments, strengthened the correlation between the occurrence of type 4 bacteroids and acetylene reduction. A new nomenclature is proposed to distinguish the histological areas in alfalfa nodules which account for and are correlated with the multiple stages of bacteroid development.     

A histological study of the organogenesis of the digestive system in bay snook Petenia splendida Günther, 1862 from hatching to the juvenile stage

In bay snook (Petenia splendida) larvae the histological development of the digestive system and swim bladder, and their relative timing of differentiation were studied from hatching to 45 days post‐hatch (dph) at 29°C. Newly hatched larvae showed a simple digestive tract, which appeared as a straight undifferentiated tube lined by a single layer of columnar epithelial cells (future enterocytes). The anatomical and histological differentiation of the digestive tract and accessory glands was a very intense, asynchronous process, proceeding from the distal to the anterior part. The intestine was the first region to differentiate (9 days post‐hatch – dph, 6.5 mm SL), and the oesophagus the last (21 dph, 8.4 mm SL). At the onset of feeding, the digestive system was organized into different functional and histologically differentiated sections, such as the buccopharynx, oesophagus, glandular stomach, and anterior and posterior intestine. This organization resembled that of the juveniles, with the exception of pharyngeal teeth and buccopharyngeal as well as oesophageal goblet cells, which proliferated later during the mixed feeding period. Histological observations revealed that bay snook larvae retained endogenous yolk reserves until 24 dph (8.9 ± 0.4 mm SL), which might be helpful for weaning this species onto a compound diet. The important lipidic accumulation observed in the intestinal mucosa, liver, and pancreas in fish fed a compound trout diet indicated that although fish were able to digest and absorb lipids, the diet formulation did not fit the nutritional requirements of early juveniles of this species. The ontogeny of the digestive system followed the same general pattern as in most cichlid species described to date. However, we detected species‐specific differences in the timing of differentiation that were related to their reproductive guild. According to the histological results, some recommendations regarding the intensive culture of this species are also provided.     

Evidence of testicular dysgenesis syndrome in the dog

Human male reproductive disorders comprising testicular dysgenesis syndrome (TDS) have become more prevalent during the last 50 years. These disorders include cryptorchidism, hypospadias, decreased semen quality, and the development of seminomas. Based on experimental evidence, it has been suggested that environmental pollutants with oestrogen-like or anti-androgenic activities play a role in the pathogenesis of TDS. In humans, histological lesions associated with TDS have been well characterized; this includes seminomas as well as their precursors, carcinoma in situ (CIS) lesions. CIS are seminiferous tubules lined by gonocytes and are a sign of testicular maldevelopment. Such CIS have recently been described in canine species, and an increased frequency of testicular tumours in dogs has also been reported. In this study, we investigated the presence of TDS lesions in canine testes submitted to routine histological examination. Histological features considered typical of human TDS were observed in 8/38 dogs examined; as in humans, individual dogs presented with various TDS features with a range of severity. In all eight dogs, CIS and at least one of the histological feature of TDS was observed in combination with seminoma. These findings suggest that as in humans, TDS may predispose canines to develop testicular cancer. A larger study is needed to better evaluate the actual incidence of TDS in canines, its clinical consequences, and the possible underlying pathogenic factors.     

Reproductive biology of the shore crab Carcinus maenas (Decapoda,Portunidae): a macroscopic and histological view

A histological study of the reproductive cycle of male and female shore crabs, Carcinus maenas (Linnaeus), was performed monthly on the South West coast of Ireland (from December 2006 to July 2008). The calculated sex ratio deviated from equality, 1:0.53, revealing a strong male bias. A system was devised, based on screening of tissue sections, to describe and stage gametogenic development. Histological examinations revealed that ovarian development occurred biannually, with a primary winter cycle in which the larger crabs reproduced and a secondary summer cycle, when smaller crabs reproduced. An association was observed where more of the larger specimens were caught in the summer months and the smaller specimens in the winter months, which inversely correlated with the segregated breeding cycles. There was strong evidence that mature male crabs could potentially copulate year round since all mature specimens, collected throughout the year, contained viable spermatozoa. Developmental stages of oogenesis and spermatogenesis were described to develop a practical gonadal index for this portunid crab, providing information on the biology of this species, which will be of benefit for fisheries management.