Effect of anaerobic digestion temperature on odour, coliforms and chlortetracycline in swine manure or monensin in cattle manure.

AIMS: This study evaluated the effect of anaerobic digestion at 22, 38 and 55°C on odour, coliforms and chlortetracycline (CTC) in swine manure or monensin (MON) in cattle manure. METHODS AND RESULTS: Swine or cattle were fed the respective growth promotant, manure was collected, and 2-l laboratory methane digesters were established at the various temperatures and sampled over 25 or 28 days. After 21 days, the concentration of CTC in the 22, 38 and 55°C swine digester slurries decreased 7, 80 and 98%, respectively. Coliforms in the 22°C digester slurries were still viable after 25 days; however, they were not detectable in the 38 and 55°C slurries after 3 and 1 days, respectively. After 28 days, the concentration of MON in the 22, 38 and 55°C cattle digester slurries decreased 3, 8 and 27%, respectively. Coliforms in the 22°C cattle digester slurries were still viable after 28 days; however, they were not detectable in the 38 and 55°C slurries after 14 and 1 days, respectively. CONCLUSIONS: These studies indicate that anaerobic digestion at 38 or 55°C may be an effective treatment to reduce coliforms and CTC; however, it is not an effective treatment to reduce MON. SIGNIFICANCE AND IMPACT OF THE STUDY: More studies are needed to determine which pharmaceuticals are susceptible to degradation by a specific manure treatment to prevent negative environmental consequences.

Collective danger and individual risk: cultural perspectives on the hazards of medical research.

Changes in medical research ethics in the past two decades have made the communication of risk to potential participants a legal imperative. Using ethnographic data from two different cultures, we examine the hazards associated with medical research in relation to the respective societal contexts that imbue them with meaning. The Iban, a Dayak people indigenous to Borneo, perceive the hazards of participating in research in terms of danger to the collective. In Australia they are construed in terms of risk to individuals. Risk in medical research is one manifestation of a broader notion of 'risk' that is constitutive of the research enterprise itself and, we argue, fundamental to post-industrial society.

Water quality in cattle feedyard playas in winter and summer.

OBJECTIVE: To determine the impact of feedyards on endotoxin concentration, fecal coliform count, and other water quality measurements during winter and summer in feedyard playas (shallow lakes). SAMPLE POPULATION: Water samples obtained from 7 feedyard playas and 3 nonfeedyard control playas. PROCEDURE: Surface water samples were collected from each playa and at various depths from 3 feedyard playas. Endotoxin concentrations, 22 water quality variables, and fecal coliform counts were determined in samples collected in summer and winter from various combinations of playas. RESULTS: Cattle numbers per feedyard ranged from 40,000 to 175,000 head/y. Mean endotoxin concentrations were significantly lower in control playas than in feedyard playas in winter and summer. Endotoxin concentration appeared to be homogenous at various water depths. Values for 20 of 22 water quality variables were higher in the feedyard playas than in control playas in winter and summer. In winter only, mean total fecal coliform concentration in feedyard playas was significantly greater than in control playas. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that feedyards have the potential to impact water quality in playas, and cattle should not be allowed access to them. Feedyard playa water should not be used under high pressure to settle dust in pens with cattle or to cool cattle, because aerosols containing pathogens and high concentrations of endotoxin are a health hazard for humans and cattle?

Exon skipping in the gene encoding pituitary adenylate cyclase-activating polypeptide in salmon alters the expression of two hormones that stimulate growth hormone release.

In mammals, GRF and pituitary adenylate cyclase-activating polypeptide (PACAP) are encoded in separate genes. We report here that in the salmon a 4.5-kilobase gene contains five exons that encode the biologically active part of the GRF-like peptide (amino acids 1-32) on exon 4 and PACAP on exon 5. Analysis of two fish messenger RNAs reveals that a long precursor containing GRF and PACAP and a short precursor containing only PACAP are both expressed in the brain of at least five species of salmon, whereas mice express only the long precursor encoded by the PACAP gene. Synthetic salmon PACAP-38 and salmon GRF-like peptide-45 both stimulated GH release from cultured salmon pituitary cells; PACAP stimulated a concentration-dependent release of GH at both 4 and 24 h of incubation, whereas GRF-like peptide did not. Alternative splicing, resulting in the short precursor in which GRF-32 is excised, may provide a means for differential control of GH secretion with higher production of the more potent PACAP. A duplication of the GRF-like/PACAP gene in evolution after the divergence of fish and tetrapods would explain separate genes and regulation for GRF and PACAP in mammals.

Sequence and expression of cDNA for pituitary adenylate cyclase activating polypeptide (PACAP) and growth hormone-releasing hormone (GHRH)-like peptide in catfish.

Growth hormone-releasing hormone (GHRH) and pituitary adenylate cyclase activating polypeptide (PACAP) are two neuropeptides that are associated with the release of pituitary growth hormone. Here a cDNA of 2501 base pairs encoding both a PACAP and a GHRH-like peptide was isolated from a brain cDNA library made from Thai catfish (Clarias macrocephalus). The organization is unlike that of the mammalian gene where PACAP and PACAP-related peptide (PRP) are encoded in one gene, and the GHRH peptide is on a separate gene. Northern analysis of catfish brain mRNA indicated that PACAP/GHRH-like mRNA has three sizes; bands of 6000, 2500, and 1000 bases suggest alternative splicing of the gene. Reverse transcriptase/PCR assay detected PACAP/GHRH-like mRNA in tissues from the brain, testis, ovary, and stomach, but not from the pancreas, pituitary, muscle, and liver. Our hypothesis that the two mammalian genes encoding GHRH or PACAP originated from a gene duplication between fish and tetrapods is supported by the present findings of similar mRNA organization and pattern of expression for the one fish gene and two mammalian genes.

Two salmon neuropeptides encoded by one brain cDNA are structurally related to members of the glucagon superfamily.

A cDNA that codes for two peptides in the glucagon superfamily has been isolated from sockeye salmon brain. The first peptide is related to growth hormone-releasing hormone (GHRH), which has high sequence similarity with PACAP-related peptide. The second peptide is structurally related to vasoactive intestinal peptide, which is also related to a newly identified peptide in mannals, pituitary adenylate-cyclase-activating polypeptide (PACAP). The salmon precursor contains 173 amino acids and has dibasic and monobasic enzyme-processing sites for cleavage of a 45-amino-acid GHRH-like peptide with a free C-terminus and a 38-amino-acid PACAP with an amidated C-terminus. The salmon GHRH-like peptide has 40% amino acid sequence identity with a human GHRH and 56% identity with human PACAP-related peptide. The 38-amino-acid salmon PACAP is highly conserved (89-92% identity) with only three or four amino acid substitutions compared with the human, ovine and rat 38-amino-acid PACAP. Not previously reported for mammalian species, a short precursor coding for only one peptide exists in salmon in addition to the long precursor coding for two peptides. In the short precursor, the coding region for GHRH is deleted leaving the PACAP-coding region in a correct reading frame. This provides one possible control mechanism for an increased expression of one peptide (PACAP) without the concomitant increase in the other peptide (GHRH) as occurs in a double-peptide precursor. The importance of the 3' non-translated region of the salmon GHRH/PACAP precursor in the regulation of translation is suggested by its 70% nucleotide sequence identity to the 3' non-translated regions of the mammalian PACAP precursors. The structural organization of the salmon GHRH/PACAP precursor provides a possible evolutionary scheme for precursors that contain tandem peptides in the glucagon superfamily.

Primary structure of neuropeptide Y from brains of the American alligator (Alligator mississippiensis).

The purification of NPY from brains of the American alligator (Alligator mississippiensis) was achieved using reverse-phase high performance liquid chromatography (HPLC). The amino acid sequence was determined using automated Edman degradation as Tyr-Pro-Ser-Lys-Pro-Asp-Asn-Pro-Gly-Glu- Asp-Ala-Pro-Ala-Glu-Asp-Met-Ala-Arg-Tyr-Tyr-Ser-Ala-Leu-Arg-His-Tyr-Ile- Asn-Leu - Ile-Thr-Arg-Gln-Arg-Tyr. Alligator NPY is the first non-mammalian vertebrate to have 100% sequence identity to human NPY. The conservation of alligator NPY suggests that serine in position 7 of chicken NPY evolved after the birds and reptiles diverged from a common Archosaurian ancestor. Furthermore, the sequence identity between alligator and human NPY suggests this sequence is the same as the ancestral amniote NPY.

Primary structure of two forms of gonadotropin-releasing hormone from brains of the American alligator (Alligator mississippiensis).

Two forms of gonadotropin-releasing hormone (GnRH) have been purified from brains of the American alligator, Alligator mississippiensis, using reverse-phase high-pressure liquid chromatography (HPLC). The concentration of total GnRH was 8.8 ng/g of frozen brain tissue or 21.1 ng per brain. The amino acid sequence of each form of GnRH was determined using automated Edman degradation. The presence of the N-terminal pGlu residue was established by digestion studies with bovine pyroglutamyl aminopeptidase and coelution with synthetic forms of the native peptide. The primary structure of alligator GnRH I is pGlu-His-Trp-Ser-Tyr-Gly-Leu-Gln-Pro-Gly-NH2 and alligator GnRH II is pGlu-His-Trp-Ser-His-Gly-Trp-Tyr-Pro-Gly-NH2.

Evidence of a growth hormone-releasing hormone-like molecule in salmon brain, Oncorhynchus keta and O. kisutch.

An antiserum with conformational specificity to human growth hormone-releasing hormone (hGH-RH 1-44 NH2) was produced and used to develop a radioimmunoassay to detect immunoreactive (ir) GH-RH in brain extracts of salmon, guinea pig, and mouse. Evidence of an immunoreactive GH-RH from salmon brain extracts with a retention time on reverse-phase high-performance liquid chromatography (HPLC) distinct from hGH-RH 1-44 is presented. Salmon irGH-RH from crude extracts elutes 1-2 min earlier than hGH-RH 1-44 (NH2) in a gradient HPLC system, whereas affinity-purified salmon irGH-RH elutes 12-13 min earlier in a near-isocratic system, suggesting that the salmon molecule is more hydrophilic.

Neuropeptide families: an evolutionary perspective.

Changes in the structure and function of five neuropeptide families during evolution are considered. The families of gonadotropin-releasing hormone (GnRH), corticotropin-releasing factor (CRF), growth hormone-releasing hormone (GH-RH), somatostatin (SS), and vasopressin/oxytocin (VP/Oxy) are used as models to illustrate the importance of a phylogenetic approach in understanding neuropeptide structure/activity relationships, precursors, processing, gene duplication, novel locations and functions, and gene-associated peptides.

Effects of pH and/or calcium-enriched freshwater on plasma levels of vitellogenin and Ca2+ and on bone calcium content during exogenous vitellogenesis in rainbow trout (Salmo gairdneri).

1. Three year old rainbow trout were exposed to low pH (5.1) and/or calcium-enriched (1.52 mM) freshwater for 10 weeks. 2. Plasma was collected periodically from individually-marked fish for analysis of total calcium and alkaline-labile phosphate (vitellogenin). 3. After the last sample gonadosomatic and hepatosomatic indices were measured and the caudal vertebrae centra were analysed for total calcium content. 4. Female trout exposed to calcium-enriched freshwater had increased plasma vitellogenin levels compared to females in soft water, whereas there was a tendency for low pH to decrease plasma vitellogenin in these fish. 5. The gonadosomatic and hepatosomatic indices were reduced in female trout exposed to acidified water. 6. There was no evidence of bone demineralization in trout exposed to low pH.

The effects of low pH on egg and alevin survival of kokanee and sockeye salmon, Oncorhynchus nerka.

1. The effects of low pH water on embryogenesis and vitellogenesis in kokanee and sockeye salmon (Oncorhynchus nerka) were investigated. Eggs were exposed to low pH from fertilization to 45 days post-median hatch or to an episodic exposure at pH 4.0. Adult kokanee were also exposed to low pH just prior to ovulation and spawning. 2. The most sensitive stages of development during chronic or episodic exposure to low pH were early embryonic development and newly-hatched alevins. 3. Incubation of eggs at low pH caused a lower median survival, delayed hatching, higher alevin mortality and reduced the efficiency of yolk conversion to tissue of yolk-sac alevins. Those effects were more pronounced when the eggs were fertilized at low pH. 4. Exposure of sexually mature kokanee salmon to acidified water reduced egg and alevin survival, delayed embryo hatching and decreased the percent hatch. Those effects were more pronounced when their eggs were incubated at low pH.

The effect of pH and/or calcium-enriched freshwater on gill Ca2+-ATPase activity and osmotic water inflow in rainbow trout (Salmo gairdneri).

Rainbow trout (Salmo gairdneri) were exposed to pH 5.0-5.1, 6.6 and/or calcium-enriched freshwater for 14 days. Hematocrit, gill Ca2+-ATPase enzyme activities, gill osmotic water inflow, plasma calcium and osmolarity were measured. No significant changes in plasma calcium ion levels were found. The typical increase in hematocrit usually associated with exposure of fish to acidified water was not found in the present study and is discussed. Plasma osmolarity decreased in fish exposed to calcium-enriched freshwater (60 mg Ca2+ X 1(-1) ) in comparison to fish exposed to control freshwater conditions (2 mg Ca2+ X 1(1) ), irrespective of the pH level. Gill Ca2+-ATPase enzyme activities were measured for both low affinity (3 mM Ca2+) and high affinity (100 microM) activity. Exposure of rainbow trout to low pH (pH 5.0-5.1) did not affect the specific activity of Ca2+-ATPase enzyme. However, low affinity Ca2+-ATPase activity in fish exposed to calcium-enriched freshwater did show a significant reduction. The increase in gill osmotic water permeability in fish exposed to calcium-enriched freshwater is interpreted as a result of the increase in osmolarity of the ambient media.