Depletion study and estimation of the withdrawal period for enrofloxacin in pacu (Piaractus mesopotamicus)

Defining the pharmacokinetic parameters and depletion intervals for antimicrobials used in fish will help in the development of important guidelines for future regulations by Brazilian agencies on the use of these substances in fish farming. This paper presents a depletion study for enrofloxacin (ENR) and its main metabolite, ciprofloxacin (CIP), in pacu (Piaractus mesopotamicus) fillets. The depletion study was carried out under monitored environmental conditions, with the temperature controlled at 27 °C to mimic the fish farming conditions in Brazil. ENR was administered orally via medicated feed for 10 consecutive days at daily dosages of 10 mg/kg body weight (b.w.). The fish were slaughtered at 6, 12, and 24 h and 2, 3, 5, 8, 12, 17, and 24 days after the medication period. Considering a maximum residue limit of 100 μg/kg for the sum of the ENR and CIP residues in the fillet, the results obtained in the depletion study allowed the estimation of a half‐life for ENR of 2.75 days and a withdrawal period of 23 days. The results obtained in this study are important for the farming of pacu in tropical regions.     

Structure and Vascularization of the Ventricular Myocardium of Piaractus mesopotamicus and Clarias gariepinus (Teleostei)

The structural characteristics of the ventricular myocardium and the arterial distribution of the ventricular wall were studied in two freshwater teleost fish species (Piaractus mesopotamicus and Clarias gariepinus). The ventricular myocardium consists of two distinct layers, an external compact layer and an internal spongy layer. In Piaractus mesopotamicus the compact layer contains fibres with an external longitudinal arrangement and an internal circular arrangement. The arterial vascularization of the compact layer is provided by subepicardial vessels and their ramifications in both species.     

Morphological,morphometric and histochemical characterization of the gastric mucosa of the camel (Camelus dromedarius)

Mucosal morphology, morphometry and mucin histochemistry of the stomach were studied in the one-humped camel. The lining of the stomach was divided into eight grossly identifiable regions. The first region was non-glandular, occupied the body of the first compartment of the stomach and constituted 53.2% of the gastric mucosa. The other seven regions were lined by a glandular mucosa. Histological, histochemical and morphometric investigations have shown that glandular mucosa comprises pseudo-cardiac, cardiac, fundic and pyloric regions. The pseudo-cardiac region was characterized by widely separated short tubular serous glands. It constituted 36.2% of the gastric mucosa; it extended over the entire lining of the second compartment and parts of the first and third compartments. The cardiac region was confined to the initial zone of the third compartment amidst the psuedo-cardiac region but contiguous with the distal end of the gastric groove. It constituted 3.4% of the gastric mucosa and was characterized by neutral and acid mucin positive glands. The fundic and pyloric regions occupied the distal distended part of the third compartment. The fundic region constituted 4.3% of the gastric mucosa. It was packed with typical fundic glands characterized by chief cells, parietal cells and acid mucin positive neck cells. The pyloric region constituted 2.9% of the gastric mucosa. Its glands were positive to acid mucins except for their bases that were positive to neutral mucins. Differences in volume densities of the mucosal components and reactivity of the surface epithelium and gastric pits to mucin stains were noted in the different regions of the stomach.     

A comparative histochemical and morphometric study of canine skeletal muscle.

The purpose of this study was to determine whether there were differences in skeletal muscle properties in the hindlimb muscles of different types of dogs. Muscle samples were obtained from the gracilis, sartorius cranial head, sartorius caudal head and tibialis anterior muscles of mixed-breed and hound-type dogs and Beagles. Fiber type, fiber size and capillary morphometry determinations of each muscle from each dog were made from sections stained for myofibrillar ATPase activity. Individual animals were bilaterally symmetric for all measured variables. Fiber type, fiber size and capillary geometry varied between dogs of a given type and muscles within a given dog. There were no differences between dog types for fiber type or fiber size; significant variation in log(muscle)/log(body) mass ratios between dog types was observed for all muscles. The results indicate that for a given muscle, significant variation can occur in skeletal muscle characteristics between different types of dogs and that these differences can be independent of differences in exercise history.     

Mixed mesodermal and mesenchymal origin of myotomal muscles in pike (Esox lucius: Teleostei)

During the myotomal myogenesis in pike (Esox lucius) two phases of muscle differentiation can be distinguished. In the first phase, the somite cells-derived stock, the primary myoblasts (of mesodermal origin), fuse to form multinucleate myotubes. Participation of myotomal cells of mesodermal origin is insufficient for further muscle development. In the second stage mesenchymal cells migrate, via myosepts, into the myotome between myotubes. Immunocytochemical detection of proliferating cell nuclear antigen (marker of S phase of cell cycle) showed their mitotic activity. Transmission electron microscope analysis revealed that the differentiation of these cells depends on their position. Cells remaining in the myosepts develop into fibroblasts and produce collagen fibres, while those that have migrated into the myotomes transform into secondary myoblasts. Mesenchymal cells in the studied species are believed to participate in hypertrophy and hyperplasy of muscle fibres. Thus the muscle fibres in pike (E. lucius) are of mesodermal-mesenchymal origin.     

Goblet Cell Types in Intestine of Tiger Barb and Black Tetra (Cyprinidae,Characidae: Teleostei)

Histochemical properties of goblet cells in intestine of a stomach‐less teleost, tiger barb (Puntius tetrazona), and a stomach‐containing teleost, black tetra (Gymnocorymbus ternetzi), are described and compared. The intestine goblet cells were mostly wide in both species, but in tiger barb, some of them were markedly thinner. In black tetra, all the intestine goblet cells displayed magenta colour after PAS, whereas in the tiger barb, only the thinner goblet cells displayed such affinity. The latter cell type was coloured strongly magenta when the tissue was treated with alcian blue (pH 2.5) followed by PAS, whereas the wide goblet cells in tiger barb and all goblet cells in black tetra displayed mainly a blue colour after such treatment. Further, the goblet cells in both species were coloured cleanly blue after high iron diamine followed by alcian blue (pH 2.5). The intestine goblet cells in both species displayed a moderate affinity to WGA and concanavalin A lectins and no affinity to DBA. Most of the goblet cells displayed no affinity to PNA, but some of them in the tiger barb displayed a moderate or strong affinity to this lectin. The affinity to WGA was somewhat strengthened after pre‐treatment with neuraminidase. These results suggest that tiger barb contains two types or variants of intestinal goblet cells: high numbers of wide cells filled by acidic, non‐sulphated mucin and some thinner cells filled by neutral mucin. The intestine goblet cells in black tetra were filled by variable amounts of neutral and acidic mucin, but the total number of such cells is much less than in tiger barb. The present lectin and neuraminidase results suggest that the intestinal mucins in both species contain significant amounts of N‐acetylglucosamine, sialic acid and glucose/mannose, but seem to lack N‐acetylgalactosamine. However, some of these cells in tiger barb contain moderate to large amounts of galactose. Together, these results suggest significant species‐specific features of the intestine goblet cells and mucin types in tiger barb and black tetra. In conclusion, the present results suggest that the diet and feeding habits in stomach‐less teleosts compared with stomach‐containing teleosts, greatly influence the number of intestine goblet cells and type of mucin in these cells.     

Differentiation and growth of myotomal muscles in a non-model tropical fish Pterophyllum scalare (Teleostei: Cichlidae)

Somite differentiation, muscle fibres formation and growth were analysed in a non-model tropical fish Pterophyllum scalare. In this study, it was found that during somite differentiation, a primary myotome appears. The primary myotome is filled with multinucleated myotubes that constitute the major part of the somite. Subsequently, Pax-3 (paired-box protein)-positive cells, located externally to the myotomes, appear. In post-hatching stages, mononucleated proliferating cell nuclear antigen-positive cells are observed in the inter-myotomal spaces and within the myotomes. The mononucleated cells, situated in the myotomes, first express desmin in their cytoplasm and then Pax-7 (paired-box protein) in their nuclei. Expression of desmin indicates that they will enter myogenic pathway, whereas expression of Pax-7 suggests their role of satellite cells. We assume that mononucleated intramyotomal cells are myogenic precursors involved in muscle growth. In advanced (post-hatching) stages of myogenesis, myotomes contain both primary and new muscle fibres. Morphometric analyses show that in Pterophyllum scalare, growth of muscle fibres is mainly a result of hypertrophy.     

Morphological, histological, and ultrastructural studies of the ovary of the cattle-tick Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae)

This study presents the morphology of the ovary, as well as the dynamics of the vitellogenesis process in oocytes of the cattle-tick Boophilus microplus. The ovary of these individuals is of the panoistic type; therefore, it lacks nurse cells. This organ consists of a single tubular structure, continuous, and composed of a lumen delimitated by a wall of small epithelial cells with rounded nuclei. In this tick species, the oocytes were classified into six stages varying from I to VI and according to: cytoplasm appearance and presence of the germ vesicle, yolk granules, and chorion. Oocytes of various sizes and at different developmental stages remain attached to the ovary through a cellular pedicel until completing stage V. Afterwards, they are liberated into the lumen and from there to the exterior. Some oocytes (classified as type VI) showed an atypical appearance indicating that some of the cellular components would be undergoing a degenerative process and/or reabsorption.     

Comparative efficacy of tricaine methanesulfonate and clove oil for use as anesthetics in red pacu (Piaractus brachypomus)

OBJECTIVE: To compare the anesthetic efficacy and physiologic changes associated with exposure to tricaine methanesulfonate and clove oil (100% eugenol). ANIMALS: 15 adult cultured red pacu (Piaractus brachypomus). PROCEDURE: Fish were exposed to each of 6 anesthetic concentrations in a within-subjects complete crossover design. Stages of anesthesia and recovery were measured, and physiologic data were collected before and during anesthesia. RESULTS: Interval to induction was more rapid and recovery more prolonged in fish exposed to eugenol, compared with those exposed to tricaine methanesulfonate. The margin of safety for eugenol was narrow, because at the highest concentration, most fish required resuscitation. Mixed venous-arterial PO2 consistently decreased with anesthesia, while PCO2 consistently increased with anesthesia in all fish regardless of anesthetic agent. The increase in PCO2 was accompanied by a decrease in pH, presumably secondary to respiratory acidosis. Anesthesia was associated with increased blood glucose, potassium, and sodium concentrations as well as Hct and hemoglobin. Fish anesthetized with eugenol were more likely to react to a hypodermic needle puncture than fish anesthetized with tricaine methanesulfonate. CONCLUSIONS AND CLINICAL RELEVANCE: Anesthesia induced with tricaine methanesulfonate or eugenol contributes to hypoxemia, hypercapnia, respiratory acidosis, and hyperglycemia in red pacu. Similar to tricaine methanesulfonate, eugenol appears to be an effective immobilization compound, but eugenol is characterized by more rapid induction, prolonged recovery, and a narrow margin of safety. Care must be taken when using high concentrations of eugenol for induction, because ventilatory failure may occur rapidly. In addition, analgesic properties of eugenol are unknown.     

The effect of testosterone on gastrocnemius muscle fibres in growing and adult male and female rats: a histochemical,morphometric and ultrastructural study

In this study, the effect of testosterone on gastrocnemius muscle fibres in growing and adult rats (male and female) was examined using histochemical, morphometric and ultrastructural techniques. After physiological saline (PS), olive oil (OvO) or olive oil + testosterone (OvOT) injections on 72 rats (growing and mature, 36 male and 36 female), the sample tissues of fibre types of the gastrocnemius muscle taken were examined by histochemical [alkaline adenosine triphosphatase (alk-ATPase), acid ATPase (ac-ATPase)], morphometric and ultrastructural techniques. In PS-injected control groups, the gastrocnemius muscle of both sexes contained all the fibre types studied [slow-oxidative muscle fibres (type I), fast-oxidative glycolytic muscle fibres (type IIA) and fast-glycolytic muscle fibres (type IIB)]. The type I fibres had the smallest diameter, type IIA had a medium diameter and type IIB fibres had the largest diameter. In OvO-injected groups, it was observed that the OvO had little effect on the gastrocnemius muscles of either sex, although there was significant enlargement of type IIB fibres. After the injection of OvOT, hypertrophy of muscle fibres was determined by morphometric study. The biggest increase in diameter was on type I fibres. In addition, degenerations on some mitochondria, accumulation of lipid droplets on type I and type II fibres, an increase in glycogen particles, bifurcation of myofibrils, an increase in the number and diameter of units resembling T tubules and an increase in ribosomal content were also observed in the same group by transmission electron microscope. Consequently, it was determined that testosterone can induce protein synthesis in gastrocnemius muscle fibres, and induces changes in shape and size, and also can change the appearance and the number of fibres.     

Morphological and morphometric features of knee joint in wild boar (Sus scrofa)

In the study, it was aimed to reveal the structure of the knee joint anatomically and morphometrically in the wild pig known as the ancestor of the domestic pig. In the study, the left and right knee joints of mature 10 wild boars were used as material. For this purpose, the materials were fixed with 10% formaldehyde solution. Macroanatomical and morphometrical examinations of bones, menisci and ligaments involved in joint formation were performed after fixation. Trochlea and condyles of femur were examined, and various measurements were made. Trochlear labia were found to be the same size, and the condyles did not show a significant difference in size. The shape of lateral condyle of the tibia was trapezoid, and the shape of medial condyle was triangular. Moreover, the lateral intercondylar tubercule was higher according to the medial one. The presence of a shallow groove on the tibial tuberosity was detected. It was found that the width difference between apex and base was more pronounced on the anterior and lateral surface of the patella. Joint surface of the fibula was found to be flat in all materials. The menisci were ‘C’ shape, but the medial meniscus is more convex than the lateral one. It was observed that the outer edges were thick, while the inner edges were thin. Ligaments of the knee joint were well developed. When the findings of the study were compared with the literature, domestication seemed to have no significant effect in terms of the morphologic properties of the knee joint of wild boar.     

Anatomical, histological and morphometric data on the urethral muscle (musculus urethralis) of the male calf

The gross and microscopic anatomy and the morphometry of the Musculus urethralis in the male calf Parts of the male urethra were examined topographically and microscopically in the male calf. The thickness and the straight course of the fibres of the urethral muscle could best be analyzed in the ventromedian region of the urethra. The results produced data on the surface area and the distribution of the two primary metabolic fibre types. This showed that the m. urethralis is capable of contracting quickly with predominantly anaerobic metabolism. The different relationships between type I and type II fibres could be determined also in relation to the position of the disseminate part of the prostate.     

Morphological,histological and histochemical studies of the gonads of ovine freemartins

Freemartins are XX/XY chimaeras that develop as a result of the fusion of the placental circulation of at least one male and one female fetus. Of 19 freemartin ewes, 13 had testis-like structures, seven of them in an abdominal position and six in an inguinal position. Histologically, their gonads had structures resembling seminiferous tubules and interstitial cells, and grossly, most had structures derived from the mesonephric ducts (vasa deferentia, epididymides and vesicular glands). The other six freemartin ewes had small, undifferentiated gonads that lacked either follicles or seminiferous tubule-like structures. They also lacked any structures derived from the mesonephric ducts. No derivatives of the paramesonephric ducts were detectable in any of the freemartin ewes. The gonads of the male-type freemartins stained immunocytochemically for 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and histochemically for alkaline phosphatase (AP) in a similar way to, but more extensively and intensely than, the gonads of normal rams, and the staining was confined to interstitial cell-like structures. The staining in the undifferentiated-type freemartins was weak, but both 3beta-HSD and AP were present in unidentified cell types.     

Geometric morphometric study of the skull shape diversification in Sciuridae (Mammalia,Rodentia)

It is generally accepted that the high phenotypic diversity of mammals is a combined result of developmental constraint and ecological adaptation, although the influence of these endogenous and exogenous factors varies in different mammal groups. The rodent family Sciuridae represents an ideal candidate for examining phenotypic diversity in relation to phylogeny and ecological adaptations. In the present study, we investigate the effects of phylogeny and lifestyle on the skull shape in different species of Sciuridae by applying geometric morphometric methods. In addition, we investigate the importance of allometry on sciurid skull shape, because results from geometric morphometrics sometimes dispute those of traditional morphometry. Here, we identify significant associations between patristic distances obtained from molecular phylogeny and shape distances in all 3 views of the cranium and the lateral view of the mandible. Multivariate regression demonstrates that shape differences among lifestyle categories are substantial, especially in the dorsal and ventral structures after the influence of phylogeny is taken into account. Allometry plays an important role in the shape variation, although its importance on different skull structures varies. Our results indicate that complex structures of this highly diverse mammal group, which occupies different niches, are affected by ecological factors and developmental constraint.     

Pharmacokinetics of enrofloxacin in the red pacu (Colossoma brachypomum) after intramuscular, oral and bath administration

The intramuscular (i.m.), oral (p.o.), and bath immersion disposition of enrofloxacin were evaluated following administration to a cultured population of red pacu. The half-life for enrofloxacin following i.m. administration was 28.9 h, considerably longer than values calculated for other animals such as dogs, birds, rabbits, and tortoises. The 4 h maximum concentration ( C _max) of 1.64 μg/mL following a single 5.0 mg/kg dosing easily exceeds the in vitro minimum inhibitory concentration (MIC) for 20 bacterial organisms known to infect fish. At 48 h post i.m. administration, the mean plasma enrofloxacin concentration was well above the MIC for most gram-negative fish pathogens. The gavage method of oral enrofloxacin administration produced a C _max of 0.94 μg/mL at 6–8 h. This C _max was well above the reported in vitro MIC. A bath immersion concentration of 2.5 mg/L for 5 h was used in this study. The C _max of 0.17 μg/mL was noted on the 2 hour post-treatment plasma sample. Plasma concentrations of enrofloxacin exceeded published in vitro MIC's for most fish bacterial pathogens 72 h after treatment was concluded. Ciprofloxacin, an active metabolite of enrofloxacin, was detected and measured after all methods of drug administration. It is possible and practical to obtain therapeutic blood concentrations of enrofloxacin in the red pacu using p.o., i.m., and bath immersion administration. The i.m. route is the most predictable and results in the highest plasma concentrations of the drug.