Fake spawns and floating particles: a rebuttal of Karkarey et al. “Alternative reproductive tactics and inverse size-assortment in a high-density fish spawning aggregation”

Courtship and spawning behaviors of coral reef fishes are very complex, and sufficient sampling effort and proper methods are required to draw informed conclusions on their mating systems that are grounded in contemporary theories of mate choice and sexual selection. We reviewed the recent study by Karkarey et al. (BMC Ecol 17:10, 2017) on the spawning behavior of Squaretail coralgrouper (Plectropomus areolatus) from India and found no evidence to support their findings of alternative reproductive tactics, unique school-spawning involving a single male with multiple females, or inverse size-assortment. The study lacks scientific credibility due to a lack of rigor in the methodology used, misinterpretation of observed behaviors, misinterpretation of the literature, and insufficient data. Their approach led the authors to produce spurious results and profound, invalid conclusions that violate the most basic assumptions of mate choice and sexual selection theory as applied to mating systems in marine fishes.     

Supplements of transgenic malt or grain containing (1,3-1,4)-beta-glucanase increase the nutritive value of barley-based broiler diets to that of maize

1. A diet with addition to normal barley of malt from transgenic barley expressing a protein engineered, thermotolerant Bacillus (1,3-1,4)-beta-glucanase during germination has previously been demonstrated to provide a broiler chicken weight gain comparable to maize diets. It also reduced dramatically the number of birds with adhering sticky droppings, but did not entirely eliminate sticky droppings. One of the objectives of the broiler chicken trials reported here was to determine if higher concentrations of transgenic malt could alleviate the sticky droppings. 2. Another aim was to investigate the feasibility of using mature transgenic grain containing the thermotolerant (1,3-1,4)-beta-glucanase as feed addition and to compare diets containing transgenic grain to a diet with the recommended amount of a commercial beta-glucanase-based product. 3. Inclusion of 75 or 151 g/kg transgenic malt containing 4.7 or 98 mg/kg thermotolerant (1,3-1,4)-beta-glucanase with 545 or 469 g/kg non-transgenic barley instead of maize yielded a weight gain in Cornish Cross broiler chickens indistinguishable from presently used maize diets. The gene encoding the enzyme is expressed in the aleurone with a barley alpha-amylase gene promoter and the enzyme is synthesised with a signal peptide for secretion into the endosperm of the malting grain. 4. Equal weight gain was achieved, when the feed included 39 g/kg transgenic barley grain [containing 66 mg/kg thermotolerant (1,3-1,4)-beta-glucanase] and 581 g/kg non-transgenic barley instead of maize. In this case, the gene encoding the enzyme has been expressed with the D-hordein gene (Hor3-1) promoter during grain maturation. The enzyme is synthesised as a precursor with a signal peptide for transport through the endoplasmic reticulum and targeted into the storage vacuoles. Deposition of the enzyme in the prolamin storage protein bodies of the endosperm protects it from degradation during the programmed cell death of the endosperm in the final stages of grain maturation and provides extraordinary heat stability. The large amount of highly active (1,3-1,4)-beta-glucanase in the mature grain allowed the reduction of the transgenic grain ingredient to 0.2 g/kg diet, thus making the ingredient comparable to that of the trace minerals added to standard diets. 5. A direct comparison using transgenic grain supplement at the level of 1 g/kg of feed with the standard recommended addition of the commercial enzyme preparation Avizyme 1100 at 1 g/kg yielded equal weight gain, feed consumption and feed efficiency in birds fed a barley-based diet. 6. The production of sticky droppings characteristic of broilers fed on barley diets was avoided with all 9 experimental diets and reduced to the level observed with a standard maize diet by supplementation with transgenic barley. 7. The excellent growth and normal survival of the 400 broilers tested on barley diets supplemented with transgenic grain or malt showed the grain and malt not to be toxic. 8. The barley feed with added transgenic grain or malt containing thermotolerant (1,3-1,4)-beta-glucanase provides an environmentally friendly alternative to enzyme additives, as it uses photosynthetic energy for production of the enzyme in the grain and thus avoids use of non-renewable energy for fermentation. The deposition of the enzyme in the protein bodies of the grain in the field makes coating procedures for stabilisation of enzyme activity superfluous. 9. Barley feed with the small amount of transgenic grain as additive to normal barley provides an alternative for broiler feed in areas where grain maize cannot be grown for climatic reasons or because of unsuitable soil and thus has to be imported.     

Mucinous vacuolar change of porcine urothelium induced by regional embolism or cryoablation

Porcine urinary tract epithelium responds to systemic infections with an increase in mucin secretion that has been called "mucinous degeneration." Here we describe similar changes in calyceal, pelvic, and ureteric urothelium in pig renal tracts 14 days after local embolism and cryoablation (four kidneys) or cryoablation alone (six kidneys). Large mucin-filled vacuoles surrounded by smaller cytoplasmic vesicles were present in the affected urothelium but were rare in calyceal urothelium of normal kidneys. These mucinous vacuolar changes were proportional to the extent of renal necrosis and could be important in the pathologic responses of xenotransplanted pig kidneys.     

S-adenosyl-L-methionine (SAMe) for the treatment of acetaminophen toxicity in a dog

An 8-month-old, spayed female Shetland sheepdog presented 48 hours after ingesting acetaminophen (1 gm/kg body weight). On presentation, the dog was laterally recumbent and hypovolemic. The dog had brown mucous membranes, severe Heinz-body hemolytic anemia, bleeding tendencies, and a red blood cell (RBC) glutathione (GSH) concentration that was 10% of reference values, despite a regenerative erythroid response. Treatment with s-adenosyl-l-methionine (SAMe) as a GSH donor successfully rescued this dog, despite the animal's late presentation after drug ingestion. A loading dose (40 mg/kg body weight) of a stable SAMe salt per os was followed by a maintenance dose (20 mg/kg body weight) sid for 7 days. Additional therapeutic interventions included an intravenous (i.v.) infusion of one unit of packed RBCs (on admission), i.v. fluid support (3 days), and famotidine (7 days) to reduce gastric acidity. Sequential assessment of RBC GSH concentrations and RBC morphology documented response to antidote administration within 72 hours. This case suggests that SAMe may provide a therapeutic option for treatment of acetaminophen toxicosis in dogs capable of retaining an orally administered antidote and maintaining adequate hepatic function for metabolism of SAMe to its thiol substrates.     

Toxic hepatic failure in newborn foals

Eight foals, 2 to 5 days of age, with similar clinical signs and laboratory and pathologic findings, died from hepatic failure. The predominant clinical signs were depression and icterus. Abnormally high values were found for plasma ammonia content, aromatic-to-branch-chain amino acid ratio, total serum bilirubin content, gamma glutamyl transferase activity, alkaline phosphatase activity, and PCV; partial thromboplastin time and prothrombin time were prolonged. Some foals had high sorbitol dehydrogenase activity. These laboratory findings were suggestive of subacute hepatic disease and failure. Predominant pathologic findings were limited to the liver and brain. The livers were less than half the expected size for 2- to 5-day-old foals, had prominent bile ductule proliferation, hepatic cell necrosis, and mild periportal fibrosis. These findings suggested both prenatal and postnatal diseases caused by exposure to a hepatoxin. The predominant lesion in the brain was the presence of Alzheimer type II astrocytes, which are characteristic of hepatoencephalopathy. Although the periportal fibrosis was suggestive of in utero exposure to a toxin, epidemiologic information suggested that the hepatic failure more likely resulted from oral inoculation of a microorganism culture product at birth. The same disease was reproduced in 2 newborn foals by feeding this product.     

Rheological properties and baking performance of new oat beta-glucan-rich hydrocolloids

Two new oat beta-glucan hydrocolloids (designated C-trim20 and C-trim30) obtained through a thermal-shearing process were evaluated for their potential use in food products as functional ingredients. Their rheological characteristics were investigated using steady and dynamic shear measurements. Both samples exhibited typical shear-thinning and viscoelastic properties of random coil polysaccharides. The Cross equation was also used to examine the dependence of their apparent viscosity on shear rates. Furthermore, the effects of flour replacement with C-trim20 on the physical, rheological, and sensory properties of cookies were studied. The cookies containing C-trim20 exhibited reduced spreading characteristics compared with the control due to their increased elastic properties. Also, higher water content and water activity were observed in the C-trim20 cookies. However, flour replacement with C-trim20 up to 10% produced cookies with instrumental texture properties similar to those of the control, which was in good agreement with the sensory results.     

Enhancing quality and oxidative stability of aged fried food with gamma-tocopherol

To determine the effects of gamma-tocopherol on the stability of fried food, potato chips were fried in triolein with 0, 100, or 400 ppm gamma-tocopherol. Potato chips, sampled at 1, 3, and 6 h of frying time, were aged for 0, 2, and 4 days at 60 degrees C and then evaluated for odor attributes by sensory analysis and for volatile compounds by purge-and-trap gas chromatography-mass spectrometry. Oil sampled after 1, 3, and 6 h of frying time from the fryer was evaluated for total polar compounds and retention of gamma-tocopherol. Oil extracted from the potato chips was also analyzed for residual gamma-tocopherol. gamma-Tocopherol disappeared rapidly, with only slight amounts of the original 100 ppm level detectable after the triolein was used for frying. gamma-Tocopherol significantly inhibited polar compound production in the triolein. Results showed that gamma-tocopherol inhibited the oxidation of the fried food even when only very low levels of retained gamma-tocopherol were present in the frying oil or potato chips. Nonanal formation was inhibited by gamma-tocopherol in aged potato chips. Odor analysis of the aged potato chips showed that samples with 0 ppm gamma-tocopherol had a rancid odor after being aged for 4 days. Potato chips with 400 ppm gamma-tocopherol had no rancid odors; however, as the level of gamma-tocopherol decreased in the triolein and in the potato chips, a weak plastic odor characteristic of oxidized triolein was detected.     

An evaluation of immunofluorescence and PCR methods for detection of rabies in archival Carnoy-fixed, paraffin-embedded brain tissue.

Direct immunofluorescence and PCR detection methods were compared for sensitivity in evaluating the rabies status of archival specimens of Carnoy-fixed, paraffin-embedded brain tissue. The material consisted of 23 samples obtained during a rabies outbreak in Finland in 1988, and one sample isolated from a bat researcher who died of rabies in Finland in 1985. These results were compared with the original diagnoses performed on the fresh tissues. The immunofluorescence assay detected 100% (12/12) of the rabies-positive archival cases. A PCR assay designed to detect a 139-bp target near the 5' end of the rabies nucleoprotein gene also detected 100% (12/12) of the samples identified as positive in the fresh tissue specimens. A PCR assay designed to detect a 304-bp target spanning the 139-bp target of the first assay detected only 67% (8/12) of the original cases. No false positives were recorded. Both immunofluorescence detection of antigen and PCR detection of a short region of the nucleoprotein gene are useful in determining the rabies status of fixed, paraffin embedded (archival) material.     

Gas chromatographic detection of the mycotoxin zearalenone in blood serum

A sensitive gas chromatographic method for the quantitative analysis of zearalenone in blood serum is described. Zearalenone is eluted from blood serum by column chromatography followed by base-acid extraction with dichloromethane as the organic phase. After epicoprostanol (internal standard) is added, the sample is evaporated to dryness, derivatized, and injected onto the gas chromatographic column. A number of silylating agents and reaction conditions were investigated. Derivatizing zearalenone with N-methyl-N-trimethylsilyltrifluoroacetamide in the presence of acetone at room temperature for at least 2 hr gave best results. Sensitivity limit is < 0.5 ng injected, equivalent to 100 ng zearalenone/mL blood serum. A linear standard curve is observed when 0.5-30 ng zearalenone derivative is injected onto the Perkin-Elmer gas chromatograph. For quantitation, a standard curve is prepared by plotting amounts of zearalenone (ng) injected vs. ratios for peak areas of zearalenone and epicoprostanol derivatives. The internal standard procedure improves the precision by minimizing variations in sample injections and detector response. Percent recovery from blood serum is 68-75 in the range of 1.6-8.0 micrograms zearalenone/mL blood.     

Campylobacter jejuni and Campylobacter coli inoculation of neonatal calves

Three groups of neonatal calves were inoculated orally with pathogenic strains of Campylobacter jejuni or C coli. The calves developed a mild, self-limiting enteritis characterized by thick mucoid feces. Bacteremia and fecal shedding of Campylobacter were sporadic in all inoculated calves. Two groups of calves were killed 1 to 3 weeks after inoculation to study the pathogenesis of infection. Postmortem culture of tissues revealed C jejuni or C coli most frequently in the ileum, cecum, colon, and blood. Clinical or pathologic differences between C jejuni-inoculated and C coli-inoculated calves were minimal.