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

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

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

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

Chlamydia abortus in Cows Oviducts,Occasional Event or Causal Connection?

Fifty‐seven genital tracts of regularly slaughtered culled Piedmontese cows, aged 7.4 ± 4.3 years (mean ± SD), range: 2.6–15.6 years, were grossly and microscopically examined. DNA extracted from oviducts was subjected to PCR to evaluate the presence of Chlamydia spp. The 15 PCR‐positive oviducts were subjected to Sanger sequencing and showed the presence of Chamydia abortus, with an identity range between 99 and 100%. Nine of the PCR‐positive samples belonged to the 24 animals with a normal macroscopic appearance of the whole genital tract (percentage of positive oviducts in normal genital tracts 9/24 = 37.5%), while six belonged to the 33 genital tracts with lesions in one or more organs (percentage of positive oviducts in pathological genital tracts 6/33 = 18.1%); of these, a single animal had salpingitis. The detection of C. abortus in bovine oviducts is of particular interest because it has never been previously investigated or reported.     

IRMS as a tool to obtain the carbon turnover (δ13C) in organs of weaned piglets fed glutamic acid and nucleotides

Early weaning of piglets causes stress characterized by a decrease in feed intake followed by a decline in growth rates; thus, a fast recovery represents an essential step for proper growth of these animals. Considering that IRMS is a potential tool for non‐destructive sampling and the fact that it provides time‐integrated estimate of assimilated and not just ingested nutrients turned possible its application to evaluate the effects of dietary nucleotides and glutamate on carbon turnover (δ¹³C) in organs of weanling piglets. At day 0, three piglets were slaughtered (prior to diet switch), the remaining eighty‐four piglets weaned at 21‐day‐old were randomly assigned in a complete block design with a 2 × 2 factorial arrangement of treatments (two Nu levels: 0 and 0.1% and two Glu levels: 0 and 1%), being three piglets per treatment slaughtered on trial days 3, 6, 9, 14, 21, 35 and 49. The samples were analysed by IRMS and adjusted to first‐order equation by a non‐linear regression analysis using NLIN of SAS, in order to establish exponential graphics. After that, the turnover data were submitted to analysis of variance using GLM of SAS. The turnover value (t_95%) verified for spleen was faster (p < 0.05) when glutamate was supplemented in diets. For pancreas and liver, the turnover rates were faster (p < 0.05) for the mixture of additives. However, for renal tissue, the turnover rate (t_95%) was greater (p < 0.05) for the free additive diet. The results obtained suggest that the mixture of additives was more efficient to develop the digestive tract at post‐weaning phase, taking into account the functional importance of pancreas and liver for nutrients’ digestion and processing.     

Chinese Box turtle (Cuora flavomarginata) with lymphoid leukemia characterized by immunohistochemical and cytochemical phenotyping

Lymphoid leukemia of T‐cell origin was diagnosed in a male Chinese Box turtle, Cuora flavomarginata, of approximately 25 years of age. The turtle presented with a history of anorexia, open‐mouth breathing, and lethargy for one week. The CBC findings included a mildly increased PCV, and severe leukocytosis due to high numbers of atypical cells interpreted to be blasts. The blasts were medium‐sized cells with round to pleomorphic nuclei, slightly clumped chromatin, indistinct nucleoli, and scant moderate‐to‐dark blue cytoplasm with occasional red‐to‐purple cytoplasmic granulation. Cytochemical and immunohistochemical staining indicated that the neoplastic cells were positive for CD3 and α‐naphthyl butyrate esterase (ANBE), leading to the diagnosis of T‐cell lymphoid leukemia. Histology of tissues collected at necropsy showed multifocal infiltrations of neoplastic round cells in the liver, spleen, kidneys, testicles, pancreas, thyroid, duodenum, bone marrow, epicardium, and myocardium. Transmission electron microscopy failed to identify viral particles within the neoplastic cells. This article describes the hematologic, histologic, and ultrastructural abnormalities associated with lymphoid leukemia in this turtle, and advanced diagnostic methods used for phenotyping the T‐cell origin.     

Effects and interaction of dietary electrolyte balance and citric acid on growth performance,intestinal histomorphology,digestive enzyme activity and nutrient transporters expression of weaned piglets

Fifty-six piglets were weaned at 21 days and randomly assigned to 1 of 8 dietary treatments with 7 replicate pens for a 14-day experimental period. The eight experimental diets were prepared via a 2 × 4 factorial arrangement with citric acid (CA; 0 and 0.3%) and dietary electrolyte balance (dEB, Na +K − Cl mEq/kg of the diet; −50, 100, 250, and 400 mEq/kg). Varying dEB values were obtained by altering calcium chloride and sodium bicarbonate contents. Dietary CA significantly increased (p < .05) villus height (VH) and villus height:crypt depth (VH:CD) in the jejunum. Piglets fed a 250 mEq/kg diet increased (p < .05) VH and VH:CD values in the duodenum. Jejunal VH and VH:CD increased (quadratic; p < .05), and ileal VH:CD (liner and quadratic; p < .05) decreased as dEB was increased in diets without CA, but no such effect was observed on the diets containing CA (dEB ×CA; p < .05). The CD in jejunum (quadratic; p < .05) increased as dEB was increased in diets containing CA, whereas it was decreased (linear; p < .05) in the diets without CA (dEB ×CA; p < .001). Dietary CA increased maltase activity and reduced the number of Ki67-positive cells (p < .05). Increasing dEB values in diets without CA increased sucrose and lactase activities (quadratic; p < .05), but no such effect was observed in the diets with CA (dEB ×CA; p < .05). An interaction effect between dEB and CA on the number of Ki67-positive cells was observed (p < .001). In conclusion, 250 mEq/kg dEB diet with CA improved piglet intestinal digestion and absorption function by improving intestinal morphology and increasing digestive enzyme activities. However, these improvements were also observed in piglets fed the 100 mEq/kg dEB diet without CA.     

Immunohistochemical study on localization of serotonin immunoreactive cells in the gastrointestinal tract of the European catfish (Silurus glanis,L.)

In this study, it was aimed to identify the distribution of serotonin immunoreactive cells within the gastrointestinal tract (GIT) of European catfish (Silurus glanis). For this purpose, the tissue samples were taken from the stomach (cardia, fundus and pylorus region) and intestine (anterior, middle and posterior region). They were examined by applying the avidin‐biotin‐immunoperoxidase method. The serotonin containing immunoreactive cells are presented in all regions of the GIT. It was determined to be localized generally in different distribution within the stomachs and intestines of S. glanis. It was found that the most intensive regions of immunoreactive cells were the cardia stomach and posterior of intestine.     

Parasites observed in the proximal alimentary tract of a Przewalski's horse in China

This article reports a Przewalski's horse from the Kalamaili Nature Reserve (KNR) in China transported to our institute to determine the cause of death. The animal had been subjected to necropsy before being transported to our institute; the entire digestive tract was removed but not opened to reduce the possible loss of parasites. During examination, ascarids in the intestines and bots in the oral cavity and stomach were observed. The parasites were identified according to an identification key and enumerated. The ascarids were identified as Parascaris spp. and were associated with a volvulus, resulting in intestinal obstruction. A total of two (stomach), one (duodenum) and 106 (intestines) Parascaris spp. were counted. Bots in the digestive tract were identified as third stage larvae of Gasterophilus pecorum (G. pecorum), Gasterophilus nigricornis (G. nigricornis) and Gasterophilus nasalis (G. nasalis), which may have been associated with stomach ulcers suffered by the horse. Attached to the lining of the oral cavity and stomach were 331 and 2014, respectively, larvae of G. pecorum, while in the duodenum there were 23 third stage larvae of G. nigricornis and 20 third stage larvae of G. nasalis. Therefore, the possibility of the presence of these parasites should not be overlooked in Przewalski horses as these parasites may be responsible for severe clinical signs in infected animals.     

Impact of mash and crumble diets on intestinal amino acids transporters,intestinal morphology and pancreatic enzyme activity of broilers

The objective of this study was to evaluate effects of mash and crumble pre‐starter diets on pancreatic enzyme activity, intestinal morphology, gene expression of intestinal peptide and amino acid (AA) transporters of broilers. Broilers in battery cages were assigned to different feed forms of pre‐starter diet from 1 to 10 days of age. Significantly increased body weight gain (BWG), feed intake (FI) and lowered FCR were observed in birds fed crumble pre‐starter diet (CPD, p < 0.05). Feed forms had no effect on whole and small intestine length, but relative intestinal length and relative small intestinal length significantly increased in the broilers fed a mash pre‐starter diet (MPD, p < 0.05). Feeding CPD increased the weight of pancreas (p < 0.05), but relative weight of the pancreas was not influenced by treatments. Pancreatic protease and amylase activities significantly increased in the broilers fed CPD (p < 0.05) but the activity of lipase was not influenced. Crypt depth (CD) and villus height (VH) were higher in broilers fed CPD (p < 0.05) but villus width (VW), villus surface area (VSA) and villus height‐to‐crypt depth ratio (VCR) were not influenced by treatments. mRNA levels for peptide transporter 1 (PepT1), Na⁺‐independent cationic AA transporter1 (CAT1), Na⁺‐independent cationic and Na⁺‐dependent neutral AA transporter 1 (y⁺LAT1) and Na⁺‐dependent neutral AA transporter (B⁰AT) were lower in birds fed CPD (p < 0.05). There were no differences in mRNA abundance of Na⁺‐independent cationic and zwitterionic AA transporter (b^0,+AT) among treatments. Overall, the present data showed that feeding crumble diet during first 10 days of age, through higher FI, enhanced intestinal histomorphology, increased digestive enzyme activity is beneficial to growth performance of broilers. Indeed, dietary form can be an important factor in the expression of jejunal transporters.     

The influence of indigestible protein on broiler digestive tract morphology and caecal protein fermentation metabolites

Indigestible dietary protein fermentation products have been suggested to negatively influence broiler performance due to their impact on health and digestive tract morphology. This study evaluated the digestive tract morphology and caecal protein fermentation metabolites of broiler fed 3 dietary protein levels (24%, 26% and 28%) with low or high indigestible protein (LIP, HIP). Two completely randomized 3 × 2 factorial trials were conducted with protein level (PL) and indigestible protein (IDP) as the main factors. In both trials, birds received six diets (24-LIP, 24-HIP, 26-LIP, 26-HIP, 28-LIP and 28 HIP) formulated with no medication. On day 5, trial 1 birds were vaccinated with Coccivac-B52, while trial 2 received no vaccine. Tissue and caecal samples were collected and caecal contents analysed for fermentation metabolites. Differences were considered significant when p ≤ .05. The LIP treatment caecal content in trial 1 at 14 days had greater histamine, agmatine and cadaverine levels, while HIP diets resulted in increased serotonin, tryptamine and spermidine. Histamine, serotonin and tryptamine at day 28 were not affected by IDP, and ammonia was not affected by treatments at day 14 or day 28. At day 14, HIP birds had lower total short-chain fatty acids, higher caecal pH and heavier pancreas, proventriculus, gizzard, jejunum and ileum weights. The same effects of IDP found in trial 1 were observed for histamine, agmatine, cadaverine, serotonin, tryptamine and spermidine at day 21 in trial 2. Trial 2 had a PL-by-IDP interaction influencing tyramine, spermidine (28-LIP > 24-LIP) and spermine with values increasing with PL for LIP diets and remaining constant for HIP diets. An interaction between PL and IDP was found for ammonia level and was similar to interactions for biogenic amines. In conclusion, dietary PL and IDP influence broiler caecal protein fermentation metabolites and those effects varied with coccidiosis vaccination and rearing environment.     

Susceptibility of Fish and Turtles to Three Ranaviruses Isolated from Different Ectothermic Vertebrate Classes

Abstract

Ranaviruses have been associated with mortality of lower vertebrates around the world. Frog virus 3 (FV3)-like ranaviruses have been isolated from different ectothermic vertebrate classes; however, few studies have demonstrated whether this pathogen can be transmitted among classes. Using FV3-like ranaviruses isolated from the American bullfrog Lithobates catesbeianus, eastern box turtle Terrapene carolina carolina, and Pallid Sturgeon Scaphirhynchus albus, we tested for the occurrence of interclass transmission (i.e., infection) and host susceptibility (i.e., percent mortality) for five juvenile fish and three juvenile turtle species exposed to each of these isolates. Exposure was administered via water bath (10³ PFU/mL) for 3 d and survival was monitored for 28 d. Florida softshell turtles Apalone ferox experienced no mortality, but 10% and 20% of individuals became infected by the turtle and fish isolate, respectively. Similarly, 5% of Mississippi map turtles Graptemys pseudogeographica kohni were subclinically infected with the turtle isolate at the end of the experiment. Channel Catfish Ictalurus punctatus experienced 5% mortality when exposed to the turtle isolate, while Western Mosquitofish Gambusia affinis experienced 10% mortality when exposed to the turtle and amphibian isolates and 5% mortality when exposed to the fish isolate. Our results demonstrated that interclass transmission of FV3-like ranaviruses is possible. Although substantial mortality did not occur in our experiments, the occurrence of low mortality and subclinical infections suggest that fish and aquatic turtles may function as reservoirs for FV3-like ranaviruses. Additionally, our study is the first to report transmission of FV3-like ranaviruses between fish and chelonians.

Received October 22, 2013; accepted January 8, 2014.     

Multidose pharmacokinetics of orally administered florfenicol in the channel catfish (Ictalurus punctatus)

Plasma disposition of florfenicol in channel catfish was investigated after an oral multidose (10 mg/kg for 10 days) administration in freshwater at water temperatures ranging from 24.7 to 25.9 °C. Florfenicol concentrations in plasma were analyzed by means of liquid chromatography with MS/MS detection. After the administration of florfenicol, the mean terminal half‐life (t_1/2), maximum concentration at steady‐state (C_ss(max)), time of C_ss(max) (T_max), minimal concentration at steady‐state (C_ss(min)), and V_c/F were 9.0 h, 9.72 μg/mL, 8 h, 2.53 μg/mL, and 0.653 L/kg, respectively. These results suggest that florfenicol administered orally at 10 mg/kg body weight for 10 days could be expected to control catfish bacterial pathogens inhibited in vitro by a minimal inhibitory concentration value of <2.5 μg/mL.     

Effects of realimentation after nutrient restriction during mid‐ to late gestation on pancreatic digestive enzymes,serum insulin and glucose levels,and insulin‐containing cell cluster morphology

This study examined effects of stage of gestation and nutrient restriction with subsequent realimentation on maternal and foetal bovine pancreatic function. Dietary treatments were assigned on day 30 of pregnancy and included: control (CON; 100% requirements; n = 18) and restricted (R; 60% requirements; n = 30). On day 85, cows were slaughtered (CON, n = 6; R, n = 6), remained on control (CC; n = 12) and restricted (RR; n = 12), or realimented to control (RC; n = 11). On day 140, cows were slaughtered (CC, n = 6; RR, n = 6; RC, n = 5), remained on control (CCC, n = 6; RCC, n = 5) or realimented to control (RRC, n = 6). On day 254, the remaining cows were slaughtered and serum samples were collected from the maternal jugular vein and umbilical cord to determine insulin and glucose concentrations. Pancreases from cows and foetuses were removed, weighed, and subsampled for enzyme and histological analysis. As gestation progressed, maternal pancreatic α‐amylase activity decreased and serum insulin concentrations increased (p ≤ 0.03). Foetal pancreatic trypsin activity increased (p < 0.001) with advancing gestation. Foetal pancreases subjected to realimentation (CCC vs. RCC and RRC) had increased protein and α‐amylase activity at day 254 (p ≤ 0.02), while trypsin (U/g protein; p = 0.02) demonstrated the opposite effect. No treatment effects were observed for maternal or foetal pancreatic insulin‐containing cell clusters. Foetal serum insulin and glucose levels were reduced with advancing gestation (p ≤ 0.03). The largest maternal insulin‐containing cell cluster was not influenced by advancing gestation, while foetal clusters grew throughout (p = 0.01). These effects indicate that maternal digestive enzymes are influenced by nutrient restriction and there is a potential for programming of increased foetal digestive enzyme production resulting from previous maternal nutrient restriction.     

Screening of in vitro nutrient digestibility coefficients of selected insect meals in broiler chickens,black-meat chickens and quails

The objective of this study is to identify potential insect species comparing with commonly used protein sources based on efficiency of the in vitro digestibility on dry matter (DMd), organic matter (OMd) and crude protein (CPd) in broiler chickens, black-meat chickens (Native breed) and quails. Each of gastric mucosa, pancreas and duodenal mucosa were obtained from proventriculus, pancreas and duodenum, respectively. Crude digestive enzyme extracts (CTE) were extracted from these organs to perform in vitro digestibility. Eighteen insect samples and six commonly used protein sources were served as substrates which were evaluated on DMd, OMd and CPd in triplicate for each substrate. The CTE from gastric mucosa was used to simulate proventriculus, whereas small intestine was simulation by adding the CTE from pancreas and duodenum. The large variation of chemical composition between insect meals was presented. For commonly used protein sources, animal proteins were higher on digestibility than plant proteins (p < .001). Quails represented a great potential to digest insect meals comparing other animals. Based on CPd results, there were potential insect species for broiler chickens (Achroia grisella: AG, Tenebrio molitor: TM and Musca domestica), black-meat chickens (Patanga succincta, TM and AG) and quails (Hermetia illucens, Acheta domesticus and Locusta migratoria; p < .001). The evidences from this study suggest that these insect species contain a great potential to use as alternative protein sources promoting an animal production with sustainability. However, the in vivo experimentation must be performed to confirm in further study.     

Effects of different protein sources completely replacing fish meal in low‐protein diet on growth performance,intestinal digestive physiology,and nitrogen digestion and metabolism in nursery pigs

The study compared the effects of selected proteins replacing fish meal in low‐protein diets on piglets’ growth performance, intestinal digestive physiology, and nitrogen digestion and metabolism. Five reduced CP, amino acid (AA)‐supplemented diets containing 4% of either S50, HP300, concentrated degossypolized cottonseed protein (CDCP), P50, or fish meal were assigned to six pens with 11 pigs for a 28‐day study period. Compared with fish meal, dietary proteins did not affect growth performance, apparent total tract digestibility (ATTD) of nutrients, serum hormone levels and biochemical parameters, apparent ileal digestibility (AID) of CP and most AA, colonic short‐chain fatty acid (SCFA) contents, duodenal and ileal morphology, digestive enzyme activity, and pH in small intestine of piglets. However, HP300, CDCP, and P50 decreased (p < 0.05) fecal N excretion per weight gain. AID of Ile in S50 and HP300 and Glu in P50 were improved (p < 0.05), and AID of Gly in other proteins was reduced (p < 0.05). S50 and P50 lowered (p < 0.05) the contents of colonic isobutyric and isovaleric. S50 and HP300 reduced (p < 0.05) jejunal villus height. CDCP increased (p < 0.05) the pepsin activity in stomach. S50, HP300, and CDCP decreased (p < 0.05) pH in the proximal colon. Overall, the selected proteins could completely replace fish meal in low‐protein diet without impairing piglets’ growth via maintaining intestinal digestive physiology, and nitrogen digestion and metabolism.     

Ontogeny of the stomach in yellow catfish (Pelteobagrus fulvidraco): detection and quantifictation of pepsinogen and H+/K+‐ATPase gene expression

Yellow catfish (Pelteobagrus fulvidraco) is an important commercial species with high aquaculture potential in China. To better understand the process of digestive functioning of gastric gland development during the larval from 1 dph (day post‐hatching) to 30 dph, real‐time PCR was used to detect and quantify the pepsinogen and H⁺/K⁺‐ATPase gene expression in P. fulvidraco. These data were also compared with the adult situation. The results showed that the expression of pepsinogen and H⁺/K⁺‐ATPase genes in P. fulvidraco larvae both started at 1 dph, though the expression level was very low until 3 dph. The quantification of pepsinogen gene expression increased significantly from 4 to 8 dph, increased fluctuantly from 8 to 23 dph and rose sharply from 23 to 30 dph. In comparison with adult fish, there were no significant differences with larvae at 5 and 23 dph. However, data of 10 and 30 dph larvae were obviously higher than those of adult group. H⁺/K⁺‐ATPase gene expression increased linearly from 1 to 30 dph. However, it was significantly lower than that of adult. The results show that P. fulvidraco larvae have an earlier functional stomach, though the function of the stomach is still not perfect. There is a gradual acidification environment within the stomach during the P. fulvidraco larvae development. Based on these results, we suggest that the weaning time for P. fulvidraco larvae would be much better after 23 dph.     

Effect of Megasphaera elsdenii NCIMB 41125 dosing on rumen development,volatile fatty acid production and blood β‐hydroxybutyrate in neonatal dairy calves

Thirty calves were randomly assigned to two treatments and fed until weaning [42 days (d) of age]. Treatments were a control group (n = 15), which did not receive Megasphaera elsdenii (Me0) and a M. elsdenii group, which received a 50‐ml oral dose of M. elsdenii NCIMB 41125 (10⁸ CFU/ml) at day 14 day of age (Me14). Calves were given colostrum for the first 3 day followed by limited whole milk feeding. A commercial calf starter was offered ad libitum starting at day 4 until the end of the study. Fresh water was available throughout the study. Feed intake and growth were measured. Blood samples were collected via jugular venipuncture to determine β‐hydroxybutyrate (BHBA) concentrations. Fourteen male calves (seven per group) were euthanised on day 42 and digestive tracts harvested. Reticulo‐rumen weight was determined and rumen tissue samples collected from the cranial and caudal sacs of the ventral and dorsal portions of the rumen for measurements of papillae length, papillae width and rumen wall thickness. Dosing with M. elsdenii NCIMB 41125 improved starter dry matter intake (DMI), weaning body weight (BW) and tended to improve average daily gain. Calves in Me14 group had greater plasma BHBA concentration than Me0‐calves during the last 3 weeks of the trial and had at day 42 greater reticulo‐rumen weight, papillae width and papillae density compared to Me0. No differences in rumen wall thickness or papillae length were observed between the two groups. Total volatile fatty acids, acetate and propionate production did not differ between treatments, but butyrate production was greater in Me14 than Me0. Dosing M. elsdenii NCIMB 41125 showed benefit for calves with improved feed intake and rumen development suggesting increased epithelium metabolism and improved absorption of digestive end products.     

Relative bioavailability of selenium yeast for broilers fed a conventional corn–soybean meal diet

The present study was conducted to assess the relative bioavailability of selenium (Se) as Se yeast (SY) relative to sodium selenite (SS) for broilers fed a conventional corn–soybean meal diet. A total of 360 one-d-old Arbor Acres commercial broilers were randomly assigned to 5 treatments with 6 replicates per treatment in a completely randomized design involving a 2 (Se sources: SY and SS) × 2 (added Se levels: 0.20 and 0.40 mg Se/kg) factorial design of treatments plus 1 (a Se-unsupplemented control diet) for 42 days. The results showed that Se concentrations in plasma, liver, heart, breast muscle, pancreas and kidney of broilers on d 21 and 42, glutathione peroxidase (GSH-Px) activity in the pancreas on d 21 as well as in the breast muscle and pancreas on d 42, and GSH-Px mRNA levels in the liver, heart, breast muscle and pancreas on d 21 increased linearly (p < .03) as levels of added Se increased. Furthermore, a difference (p ≤ .05) between SY and SS was detected for Se concentrations in plasma, liver, heart, breast muscle, pancreas and kidney, GSH-Px activity in pancreas on both d 21 and 42, as well as pancreatic GSH-Px mRNA level on d 21. Based on slope ratios from the multiple linear regressions of the above indices, the Se bioavailabilities of SY relative to SS (100%) were 111%–394% (p ≤ .05) when calculated from the Se concentrations in plasma, liver, heart, breast muscle, pancreas, kidney and GSH-Px activities in pancreas on d 21 and 42, as well as GSH-Px mRNA level in pancreas on d 21. The results from this study indicated that the Se from SY was more available for enhancing the Se concentrations in plasma or tissues and the expression and activity of GSH-Px in pancreas of broilers than the Se from SS.     

Ultrastructural characteristics of black marsh turtle spermatozoa obtained by electroejaculation

The black marsh turtle (Geoemydidae: Siebenrockiella crassicollis) is a freshwater turtle that occurs in equatorial tropical climates in South East Asia. The semen of S. crassicollis was investigated by electroejaculation. The spermatozoa of S. crassicollis are filiform in shape with curved heads. The entire length, midpiece to tail length, tail width and tail length of the spermatozoa were 71.33 ± 1.55 μm, 49.92 ± 1.13 μm, 0.43 ± 0.02 μm and 48.53 ± 0.25 μm, respectively. The head length, head width across the middle and head width across the base were 14.00 ± 0.38 μm, 0.79 ± 0.03 μm and 0.91 ±0.0.03 μm, respectively. The acrosomal region of the S. crassicollis spermatozoa was narrower than the head, with an acrosomal length and width at the annulus of 2.90 ± 0.13 μm and 0.43 ± 0.01 μm, respectively. The midpiece of the S. crassicollis spermatozoa was narrower than the head and contained 30–40 mitochondrial balls, each with a ball diameter of 0.16 ± 0.002 μm. The midpiece length, midpiece width and tail length were 4.92 ± 0.16, 0.78 ± 0.03 and 48.53 ± 0.25 μm, respectively. This study presents the characteristic appearance of a freshwater turtle spermatozoa in Southeast Asia, as observed under an electron microscope. The spermatozoa of Siebenrockiella crassicollis are morphologically different from those of other freshwater turtles from other regions described in previous studies.