Efficiency of riboflavin and ultraviolet light treatment against high levels of biofilm-derived Staphylococcus epidermidis in buffy coat platelet concentrates.

BACKGROUND AND OBJECTIVES: Staphylococcus epidermidis forms surface-attached aggregates (biofilms) in platelet concentrates (PCs), which are linked to missed detection during PC screening. This study was aimed at evaluating the efficacy of riboflavin-UV treatment to inactivate S. epidermidis biofilms in buffy coat (BC) PCs. MATERIALS AND METHODS: Biofilm and non-biofilm cells from S. epidermidis ST-10002 and S. epidermidis AZ-66 were individually inoculated into whole blood (WB) units (~106 colony-forming units (CFU)/ml) (N = 4-5). One spiked and three unspiked WB units were processed to produce a BC-PC pool. Riboflavin was added to the pool which was then split into two bags: one for UV treatment and the second was untreated. Bacterial counts were determined before and after treatment. In vitro PC quality was assessed by flow cytometry and dynamic light scattering. RESULTS: Bacterial counts were reduced during BC-PC production from ~106 CFU/ml in WB to 103 -104 CFU/ml in PCs (P < 0·0001). Riboflavin-UV treatment resulted in significantly higher reduction of S. epidermidis AZ-66 than strain ST-10002 (≥3·5 log reduction and 2·6-2·8 log reduction, respectively, P < 0·0001). Remaining bacteria post-treatment were able to proliferate in PCs. No differences in S. epidermidis inactivation were observed in PCs produced from WB inoculated with biofilm or non-biofilm cells (P > 0·05). Platelet activation was enhanced in PCs produced with WB inoculated with biofilms compared to non-biofilm cells (P < 0·05). CONCLUSION: Riboflavin-UV treatment was similarly efficacious in PCs produced from WB inoculated with S. epidermidis biofilm or non-biofilm cells. Levels of biofilm-derived S. epidermidis ≥103 CFU/ml were not completely inactivated; however, further testing is necessary with lower (real-life) bacterial levels.

Pathogen reduction of whole blood: utility and feasibility.

OBJECTIVES: To collect information on pathogen reduction applied to whole blood. BACKGROUND: Pathogen reduction (PR) of blood components has been developed over the past two decades, and pathogen-reduced fresh-frozen plasma and platelet concentrates are currently in clinical use. High cost and incomplete coverage of components make PR out of reach for low- and middle-income countries (LMIC). However, should PR become applicable to whole blood (WB), the main product transfused in sub-Saharan Africa, and be compatible with the preparation of clinically suitable components, cost would be minimised, and a range of safety measures in place at high cost in developed areas would become redundant. METHODS: All articles called with "pathogen reduction", "pathogen inactivation" and "whole blood" were retrieved from Medline. References in articles were utilised. RESULTS: One such PR technology (PRT) applied to WB has been developed and has shown efficacious against viruses, bacteria and parasites in vitro; and has been able to inactivate nucleated blood cells whilst retaining the ability to prepare components with acceptable characteristics. The efficacy of this WB PRT has been demonstrated in vivo using the inactivation of Plasmodium falciparum as a model and showing a high degree of correlation between in vitro and in vivo data. Obtaining further evidence of efficacy on other suitable targets is warranted. Shortening of the process, which is currently around 50 min, or increasing the number of units simultaneously processed would be necessary to make PRT WB conducive to LMIC blood services' needs. CONCLUSIONS: Even if not 100% effective against agents that are present in high pathogen load titres, WB PRT could massively impact blood safety in LMIC by providing safer products at an affordable cost.

Riboflavin and ultraviolet light: impact on dengue virus infectivity.

BACKGROUND: Dengue viruses (DENV 1-4) are emerging across the world, and these viruses pose a risk to transfusion safety. Pathogen inactivation may be an alternative approach for managing the risk of DENV transfusion transmission. This study aimed to investigate the ability of riboflavin and UV light to inactivate DENV 1-4 in platelet concentrates. MATERIALS AND METHODS: DENV 1-4 were spiked into buffy coat-derived platelet concentrates in additive solution (SSP+) before being treated with riboflavin and UV light. Infectious virus was quantified pre- and posttreatment, and the reduction in viral infectivity was calculated. RESULTS: All four DENV serotypes were modestly reduced after treatment. The greatest amount of reduction in infectivity was observed for DENV-4 (1·81 log reduction) followed by DENV-3 (1·71 log reduction), DENV-2 (1·45 log reduction) and then DENV-1 (1·28 log reduction). CONCLUSION: Our study demonstrates that DENV 1-4 titres are modestly reduced following treatment with riboflavin and UV light. With the increasing number of transfusion-transmitted cases of DENV around the globe, and the increasing incidence and geographical distribution of DENV, additional approaches for maintaining blood safety may be required in the future.

Development of a mitochondrial DNA real-time polymerase chain reaction assay for quality control of pathogen reduction with riboflavin and ultraviolet light.

BACKGROUND AND OBJECTIVES: Transfusion is associated with a risk of infection and alloimmunization. Pathogen reduction using riboflavin and UV light (Mirasol treatment) inactivates pathogens and leucocytes. With increasing adoption of the technology in clinical use, regulatory agencies have recommended the introduction of quality control measures to monitor pathogen reduction efficacy. We sought to develop a real-time PCR-based assay to document the impact of pathogen reduction on the mitochondrial genome in blood components. MATERIALS AND METHODS: DNA was extracted from platelet and plasma components before and after treatment with riboflavin and UV light. Inhibition of PCR amplification of mitochondrial DNA (mtDNA) in short- and long-amplicon target regions, ranging from under 200 base pairs (bp) to over 1800 bp, was measured in treated relative to untreated components. RESULTS: Pathogen reduction of platelets using riboflavin and UV light resulted in inhibition of PCR amplification of long-amplicon mtDNA targets, demonstrating approximately 1 log reduction of amplification relative to untreated products. Amplification of short-amplicon mtDNA targets was not affected by treatment. Evaluation of 110 blinded platelet samples from the PREPAReS clinical trial resulted in prediction of treatment status with 100% accuracy. Pathogen reduction of plasma components resulted in similar levels of PCR inhibition, while testing of 30 blinded plasma samples resulted in prediction of treatment status with 93% accuracy. CONCLUSION: A differential sized amplicon real-time PCR assay of mitochondrial DNA effectively documents nucleic acid damage induced by Mirasol treatment of platelets. The use of the assay for plasma product pathogen reduction requires further investigation.

Establishment of the first international repository for transfusion-relevant bacteria reference strains: ISBT working party transfusion-transmitted infectious diseases (WP-TTID), subgroup on bacteria.

BACKGROUND: Bacterial contamination of platelet concentrates (PCs) still remains a significant problem in transfusion with potential important clinical consequences, including death. The International Society of Blood Transfusion Working Party on Transfusion-Transmitted Infectious Diseases, Subgroup on Bacteria, organised an international study on Transfusion-Relevant Bacteria References to be used as a tool for development, validation and comparison of both bacterial screening and pathogen reduction methods. MATERIAL AND METHODS: Four Bacteria References (Staphylococcus epidermidis PEI-B-06, Streptococcus pyogenes PEI-B-20, Klebsiella pneumoniae PEI-B-08 and Escherichia coli PEI-B-19) were selected regarding their ability to proliferate to high counts in PCs and distributed anonymised to 14 laboratories in 10 countries for identification, enumeration and bacterial proliferation in PCs after low spiking (0·3 and 0·03 CFU/ml), to simulate contamination occurring during blood donation. RESULTS: Bacteria References were correctly identified in 98% of all 52 identifications. S. pyogenes and E. coli grew in PCs in 11 out of 12 laboratories, and K. pneumoniae and S. epidermidis replicated in all participating laboratories. The results of bacterial counts were very consistent between laboratories: the 95% confidence intervals were for S. epidermidis: 1·19-1·32 × 10(7) CFU/ml, S. pyogenes: 0·58-0·69 × 10(7) CFU/ml, K. pneumoniae: 18·71-20·26 × 10(7) CFU/ml and E. coli: 1·78-2·10 × 10(7) CFU/ml. CONCLUSION: The study was undertaken as a proof of principle with the aim to demonstrate (i) the quality, stability and suitability of the bacterial strains for low-titre spiking of blood components, (ii) the property of donor-independent proliferation in PCs, and (iii) their suitability for worldwide shipping of deep frozen, blinded pathogenic bacteria. These aims were successfully fulfilled. The WHO Expert Committee Biological Standardisation has approved the adoption of these four bacteria strains as the first Repository for Transfusion-Relevant Bacteria Reference Strains and, additionally, endorsed as a project the addition of six further bacteria strain preparations suitable for control of platelet contamination as the next step of enlargement of the repository.

The effect of pathogen reduction technology (Mirasol) on platelet quality when treated in additive solution with low plasma carryover.

BACKGROUND AND OBJECTIVES: Pathogen reduction technologies (PRT) for platelets are now compatible with both plasma and platelet additive solutions (PAS). The aim of this study was to examine the effect of PRT on the platelet storage lesion, in the presence of PAS with low plasma carryover. MATERIALS AND METHODS: PRT-treated (Mirasol) and untreated buffy coat-derived platelet concentrates prepared in 28% plasma/PAS-IIIM were evaluated using in vitro cell quality parameters on days 1, 2, 5, and 7 post-collection. RESULTS: At day 5, there were no significant differences between control and PRT treated platelets for swirl, viability, pO(2) , pCO(2) , mean platelet volume and adenosine diphosphate-induced aggregation. PRT treatment did not affect the functional integrity of the mitochondria. However, PRT resulted in a decrease in pH and enhancement of platelet glycolysis and activation, evidenced by increased glucose consumption and lactate production rates, increased expression of CD62P, CD63, annexin V staining and increased secretion of cytokines (P < 0.05). Hypotonic shock response and aggregation in response to collagen were also significantly reduced in PRT treated platelets (P < 0.05). CONCLUSION: Despite the observed differences in platelet metabolism and activation observed following PRT treatment in PAS and low plasma carryover, the results suggest that treatment and storage of platelets in PAS is no more detrimental to platelets than treatment and storage in plasma.

Characterization of plasma protein activity in riboflavin and UV light-treated fresh frozen plasma during 2 years of storage at -30 degrees C.

BACKGROUND: The Mirasol Pathogen Reduction Technology System (PRT) for Plasma (CaridianBCT) is based on a riboflavin and UV light treatment process resulting in pathogen inactivation due to irreversible, photochemically induced damage of nucleic acids. This study evaluated the in vitro protein quality of plasma products treated with riboflavin and UV light following treatment and subsequent storage for up to 104 weeks at -30 degrees C. MATERIALS AND METHODS: Apheresis and whole blood-derived plasma products were combined with riboflavin solution and exposed to ultraviolet light. Treated plasma was then flash frozen, within 8 h of collection, stored at -30 degrees C for up to 104 weeks and analysed at different stages of storage using standard coagulation assays. Results were compared with paired, untreated units stored for the same intervals. RESULTS: The average percent protein retention for all time-points in PRT-treated plasma samples after 36, 69, 87 and 104 weeks of storage at -30 degrees C in comparison with controls held under similar conditions were: Total Protein, 101%, Factor VIII, 79%, Fibrinogen, 78%, Factor II, 87%, Factor XII, 86%, Factor X, 84% and Factor IX, 81%. Anticoagulant and inhibitor proteins showed between 90% and 100% retention after 1 year (52 weeks) and 69 weeks of storage. No clinically relevant complement activation was observed in treated and stored samples. CONCLUSION: Riboflavin and UV light-treated plasma demonstrates reductions in several plasma coagulation factors following treatment. This reduction in activity levels is noted immediately after treatment and remains relatively constant during 2 years of storage at -30 degrees C.

Electron-spin domains: magnetic enhancement of superconductivity.

An inhomogeneous superconducting state, not yet conclusively identified, was predicted by Fulde and Ferrell and Larkin and Ovchinnikov (FFLO) to arise in superconductors with strong Pauli limiting, a consequence of the electrons' Zeeman (spin) energy in a magnetic field. Radovan et al. propose that the observed cascades of steps in magnetization of the heavy fermion superconductor CeCoIn5, within the recently discovered second low-temperature state, are due to transitions between Landau-level (LL) states with different m-quanta vortices, expected under certain conditions when the magnetic field is swept within the FFLO state. The authors then conclude that the observed steps in magnetization constitute a proof that the low-temperature state in CeCoIn5 is indeed an FFLO state. We argue that this interpretation of the observed steps in magnetization cannot be supported on either quantitative or qualitative grounds.

High-temperature weak ferromagnetism in a low-density free-electron gas.

The magnetic properties of the ground state of a low-density free-electron gas in three dimensions have been the subject of theoretical speculation and controversy for seven decades. Not only is this a difficult theoretical problem to solve, it is also a problem which has not hitherto been directly addressed experimentally. Here we report measurements on electron-doped calcium hexaboride (CaB6) which, we argue, show that-at a density of 7× 1019 electrons cm-3-the ground state is ferromagnetically polarized with a saturation moment of 0.07 µB per electron. Surprisingly, the magnetic ordering temperature of this itinerant ferromagnet is 600 K, of the order of the Fermi temperature of the electron gas.

Pathogen reduction technology (Mirasol) treated single-donor platelets resuspended in a mixture of autologous plasma and PAS.

BACKGROUND AND OBJECTIVES: Mirasol pathogen reduction technology (PRT) for platelet concentrates uses riboflavin and ultraviolet light. Previously, we described increased metabolism and activation for PRT platelets stored in 100% plasma. To improve platelet quality, we resuspended platelets in a mixture of plasma and platelet additive solution (PAS). MATERIALS AND METHODS: Single-donor platelets were resuspended in plasma and split into an untreated control and a PRT-treated single product. One hundred and fifty millilitre PAS (SSP+) was added to both. Over 7 days, we assayed pH, glucose consumption-, lactate production rate and CD62p with and without TRAP. RESULTS: On day 5, PRT units showed a significantly lower pH (7.087 +/- 0.105 vs. 7.288 +/- 0.200) accompanied by a higher lactate production (0.104 +/- 0014 vs. 0.063 +/- 0.017 mmol/10(12)/h) and glucose consumption rate (0.039 +/- 0005 vs. 0.028 +/- 0.009 mmol/10(12) platelets/h). CD62p expression was higher in treated units (44.5 +/- 13.0 vs. 16.5 +/- 7.6%). CONCLUSION: In comparison to PRT platelets resuspended in 100% plasma, a mixture of plasma and PAS improves pH and platelet metabolism but not platelet activation. Prolonged shelf-life for up to 7 days may be possible

Modification of lipid phase behavior with membrane-bound cryoprotectants.

Several derivatives of cholesterol containing oxyethylene headgroups with and without a terminal galactose have been synthesized in order to examine the effects of immobilizing a cryoprotectant at a membrane surface. In this work, we have studied the ability of the triethoxycholesterol (TEC) and triethoxycholesterol galactose (TEC-Gal) derivatives to modulate the phase behavior of phosphatidylcholine and phosphatidylethanolamine membranes. Methods of fluorescence polarization, 31P-NMR and freeze-fracture electron microscopy were employed to monitor these changes in lipid phase behavior. Fluorescence polarization data demonstrated the ability of the derivatives to fluidize gel state and rigidify liquid-crystalline state phosphatidylcholines in a manner similar to that observed for cholesterol. Unlike cholesterol, however, the Tm of dipalmitoylphosphatidylcholine (DPPC) was reduced in a concentration-dependent manner with each of the derivatives. Freeze-fracture electron microscopy and 31P-NMR of DOPE dispersions indicate an increase in the lamellar to hexagonal phase-transition temperature on the order of 10-20 C degrees above room temperature for mixtures with 20 mol% of the derivatives. These results are discussed in terms of the properties exhibited by compounds such as carbohydrates, which are known to serve as cryoprotectants for synthetic and biological membranes.

Photochemical and photophysical studies of 3-amino-6-iodoacridine and the inactivation of lambda phage.

The photochemistry and photophysics of 3-amino-6-iodoacridine (Acr-I) was studied. Photolysis (350 nm) of Acr-I (free base) generates products consistent with a free radical intermediate in methanol, benzene and carbon tetrachloride. The Acr-I hydrochloride is shown to bind to calf thymus DNA and to the self-complementary dinucleotide cytidylyl-(3'-5')-guanosine (CpG) miniduplex in a manner similar to that of proflavine (Acr-NH2), a known DNA intercalator. The Acr-I is shown to more efficiently nick supercoiled plasmid DNA pBR322 upon 350 nm or 420 nm photolysis than Acr-NH2. The efficiency of Acr-I-sensitized DNA nicking is not oxygen dependent. Photolysis of the Acr-I/(CpG)2 complex leads to cleavage of the dinucleotide and to cytidine base release by selective damage to a specific ribose moiety. Dinucleotide cleavage occurs equally well in the presence or absence of oxygen, thereby eliminating a singlet oxygen- or peroxyl radical-mediated process. Photolysis of Acr-I in the presence of a mononucleotide (GMP) or a non-self-complementary dinucleotide (uridylyl-[3'-5']-cytidine-UpC) does not lead to fragmentation and base release. Similarly, photolysis of the Acr-NH2/(CpG)2 complex does not lead to fragmentation and base release. The data indicate that photolysis of an iodinated intercalator bound to CpG or plasmid DNA generates an intercalated aryl radical and that the reactive intermediate initiates a sequence of reactions that efficiently nick nucleic acids. The inactivation of lambda phage sensitized by Acr-I with UV (350 nm) light is oxygen independent but with visible (420 nm) light is strongly oxygen dependent. The Acr-I fluoresces more intensely when excited at 446 than at 376 nm. Thus, UV photolysis may lead to C-I bond homolysis and free radical formation, a process that is not energetically feasible with visible light. The results demonstrate the difficulty of extrapolating model studies involving simple molecules and DNA to understanding the mechanism of viral inactivation with a particular sensitizer.

Dramatic improvements in viral inactivation with brominated psoralens, naphthalenes and anthracenes.

Amino or polyamino derivatives of naphthalene (N-H), anthracene (A-H) and 8-alkoxypsoralen (PSR-H) were prepared along with their monobrominated analogs (N-Br, A-Br and PSR-Br). The ammonium salts of these compounds are all water soluble and bind strongly to calf thymus DNA and to lambda phage, a double-helical DNA, protein-coated virus. Binding of the sensitizer to DNA occurs, presumably by a mixture of hydrophobic, intercalative and electrostatic interactions. Relative binding constants to calf thymus DNA and to lambda phage were measured by the ethidium bromide fluorescence quenching assay. In general the brominated analogs bind more tightly to calf thymus DNA and to the virus than to the nonhalogenated analogs. It is demonstrated that the brominated aromatics are much more effective at inactivating lambda phage upon photoactivation (lambda approximately 310 or 350 nm) than are their nonbrominated analogs. At identical sensitizer concentrations (by weight) and light flux N-Br, A-Br, and PSR-Br produce 5-6 more logs of viral inactivation than their nonbrominated counterparts (N-H, A-H and PSR-H, respectively). The bromine effect may originate from light-induced electron transfer and subsequent cleavage of the C-Br bond of the sensitizer radical anion bonds to form aryl radicals. Singlet oxygen cannot be responsible for the viral inactivation because the brominated sensitizers are equally effective in the presence and absence of oxygen. Dithiothreitol does not protect lambda phage from light-induced inactivation by the brominated sensitizer thereby demonstrating that the photogenerated reactive intermediates responsible for the effect are complexed to the virus and are not generated free in solution.

Temporal variability of benzene exposures for residents in several New Jersey homes with attached garages or tobacco smoke.

The U.S. Environmental Protection Agency's (EPA) previous TEAM studies of personal exposure to VOCs for 700 residents in several U.S. cities found that indoor air concentrations were often higher than outdoor levels. Several potential sources of benzene exposure were identified, including environmental tobacco smoke and materials or activities associated with attached garages. Indoor, personal, and outdoor monitoring was conducted at eleven New Jersey homes over multiple 12-hr monitoring periods. One study objective was to assess the impact of attached garages on human exposure to benzene and the variability of benzene exposure across time. Benzene was also measured in several homes inhabited by smokers and in homes without known combustion sources for comparative purposes. At homes with a garage or environmental tobacco smoke, mean indoor and personal benzene concentrations were two to five times higher than outdoor levels at all but one home. Mean personal exposures ranged from 8 to 31 micrograms/m3. Indoor/outdoor ratios were calculated and ranged from 0.8 to 11. Benzene levels in the four garages ranged from 3 to 196 micrograms/m3 and usually were higher than either indoor living areas or personal levels. Multi-zone air exchange rates were measured, and benzene source strengths in each zone were estimated. Garage source strength estimates for benzene ranged from 310 to 52,000 micrograms/h. The mass transfer of benzene from sources in the garage to home living areas was also large in three of the homes, ranging from 730 to 26,000 micrograms/h. Materials or activities in the garage were a source of benzene exposure for the residents in these three homes. Large temporal variations (factors of 2 to 30) were observed in indoor and personal benzene concentrations, indoor/outdoor ratios, and source strengths over the six or ten monitoring periods at each home. Changes in outdoor air benzene levels were an underlying factor in changing exposure levels, with indoor sources further elevating indoor air levels and personal exposures.

Influence of dietary zinc or cadmium on hair and tissue mineral concentrations in rats and goats.

Three experiments were conducted to determine whether concentrations of minerals in hair and other tissues of rats and goats are affected by level of dietary Zn or Cd. In the first experiment, rats were fed diets that contained 10.3, 20.5, 33.7, 41.3 or 52.9 micrograms Zn/g for 57 d. Rats fed the diet that contained 10.3 micrograms Zn/g suffered from mild Zn deficiency, as indicated by depressed feed intakes and slower growth rates than rats fed diets containing higher amounts of Zn. Zinc concentrations in hair (P less than .01), liver (P less than .01) and kidney (P less than .01) increased as dietary Zn increased. Confidence intervals for dietary Zn concentration predicted from Zn analysis of hair were large. In the second experiment, rats were fed diets that contained .1, 4.0, 7.6, 10.1 or 15.9 micrograms Cd/g for 57 d. Total growth, feed intake, feed efficiency and liver, kidney and testes weights were not affected (P greater than .05) by dietary Cd concentration. Cadmium increased linearly in liver (P less than .01) and kidney (P less than .01) and quadratically in testes (P less than .01) as Cd intake increased, but Cd in hair was not affected by dietary level of Cd. High correlations between Cd concentrations in liver (R2 = .88) and kidney (R2 = .90) and dietary Cd concentration indicate that Cd intakes of rats may be accurately predicted from Cd analyses of these tissues. In the third experiment, goats were fed diets containing 0, 10.4, 18.0 or 28.5 micrograms Cd/g for 125 d. Growth, feed intake, feed efficiency and liver and kidney weights were not affected by dietary Cd intake. Cadmium in hair samples was not affected by level of dietary Cd; however, cadmium in liver (P less than .01), kidney (P less than .01) and proximal duodenum (P less than .01) increased as dietary Cd increased. Cadmium in liver, kidney, lungs and proximal duodenum was highly correlated (R2 = .67, .89, .57, .49, respectively) with dietary Cd concentration.

Mineral concentrations in hair as indicators of mineral status: a review.

Mineral content of hair is affected by season, breed, hair color within and between breeds, sire, age and body location. Seasonal effects may be due to stage of growth of hair and to changes caused by perspiration, surface contamination and diet. Breed and sire effects on mineral content of hair complicate prediction of nutritional status based on hair analyses because, in many commercial cattle, neither breed nor sire is known. Hair from young animals may be lower in Zn, Mn and Fe, but is higher in Na, Ca, Cu and K than that from older animals. Pigmented hair apparently is higher in Ca, Mg, K and NA than white hair, but trace mineral concentrations are similar in hair of different colors. The effect of body location on mineral content of hair may be due to differences in surface contamination, differences in hair growth cycles and differences in texture of the hair. Concentrations of Ca, P and Cu in hair are not affected by dietary intake of these minerals. Zn and Se contents of hair may reflect dietary intake. Information on other required minerals in lacking. Pb, As and, possibly, Cd levels in hair may be related to dietary or environmental exposure. Because of the many factors that cause variation in mineral content of hair, hair analyses are not likely to be precise indicators of the mineral status of animals. Hair analyses may help to detect severe deficiencies of some required minerals or exposure to some heavy metals. However, if hair analyses are to be conducted, care must be taken to compare values from test animals with those from animals of similar breed, sex, season, sire and color. In addition, new hair growth should be analyzed, environmental contamination should be minimized and the hair samples should be cleaned before analyses.

Influence of captan on in vitro and in vivo digestibility of forage.

Four trials were conducted to examine the effects of Captan (N-trichloromethylthiotetrahydrophthalimide) on digestibility of forage. In the first trial, alfalfa-brome hay was digested in vitro with Captan added to the hay at levels ranging from 0 to 150 ppm. Digestibilities of alfalfa-brome dry matter, cell walls, acid detergent fiber, cellulose and hemicellulose exhibited a curvilinear response to Captan level. Maximum responses occurred at 75 to 100 ppm of added Captan. In the second trial, addition of 75 ppm of Captan to alfalfa-brome hay resulted in a significant increase in in vitro dry matter digestibility when fresh rumen fluid was used as the inoculum, but had no effect when distilled water or autoclaved rumen fluid was used as the inoculum. The influence of Captan on rate of in vitro digestion was also examined. Addition of 75 ppm of Captan to alfalfa-brome hay resulted in an apparent increase (19.5%) in rate of cell wall digestion. Five Angus steers were used in an experiment with a 5 x 5 Latin-square design conducted to examine the effects of Captan on in vivo digestibility. Captan was added at 0, 75, 150, 300 and 600 ppm of air dry diet. Steers fed Captan gained faster (P less than .05) and had higher (P less than .05) gains per unit of feed. Digestibilities of dry matter, cell walls, acid detergent fiber, cellulose, hemicellulose and protein were higher (P less than .01) for steers fed Captan than for those fed the control diet. There were not significant responses in gain or digestibilities as Captan level increased from 75 to 600 ppm.

Influence of supplemental nitrogen source on digestion of nitrogen, dry matter and organic matter and on in vivo rate of ruminal protein degradation.

Seven Holstein steers (340 kg) fitted with ruminal, duodenal and ileal cannulae were used to measure the influence of supplemental N source on digestion of dietary crude protein (CP) and on ruminal rates of protein degradation. Diets used were corn-based (isonitrogenous, 12% CP on a dry matter basis, and isocaloric, 80% total digestible nutrients) with urea, soybean meal (SBM), linseed meal (LSM) or corn gluten meal (CGM) as supplemental N. Ruminal ammonia N concentrations were higher (P less than .05) in steers fed LSM than in those fed CGM, but did not differ from those in steers fed urea or SBM (11.7, 6.7, 9.1 and 9.2 mg/100 ml, respectively). Due to the high degradability of urea, ruminal digestion of dietary CP was greater (P less than .05) in steers fed urea than in those fed CGM, but intermediate in steers fed SBM and LSM (58.4, 48.8, 53.1 and 53.9%, respectively). Flow of bacterial nonammonia N to the duodenum was highest (P less than .05) in steers fed SBM or LSM, intermediate (P less than .05) for urea and lowest (P less than .05) for CGM (86.8, 86.1, 76.3 and 65.9 g/d, respectively). Efficiency of bacterial protein synthesis was lowest in steers fed CGM and differed (P less than .05) from SBM (15.6 vs 21.8 g N/kg organic matter truly digested, respectively). Rate of ruminal digestion for SBM-CP differed (P less than .05) from that of CGM-CP but not from that of LSM-CP (17.70, 5.20 and 10.13%/h, respectively). The slow rate of ruminal degradability of CGM resulted in increased amounts of dietary protein reaching the intestinal tract but lower amounts of bacterial protein, thus intestinal protein supply was not appreciably altered.

Effects of compensatory growth on some body component weights and on carcass and noncarcass composition of growing lambs.

A trial was conducted in 1983 and repeated in 1984 to measure effects of restricted feed intake and realimentation on weights of organs and on carcass and noncarcass composition. A total of one hundred six weaned lambs from two breeds (Timahdit and D'man) and a breed cross (Ile de France x D'man) were used in both years. Lambs were allotted to one of six feed intake regimens: HH (ad libitum access to feed from 21 to 30 kg); HM (ad libitum access to feed from 21 to 26 kg then 70% ad libitum to 30 kg); MH (70% ad libitum from 21 to 26 kg then ad libitum to 30 kg); MM (70% ad libitum from 21 to 30 kg); LH (restricted to lose weight from 21 to 17 kg then ad libitum to 30 kg); and LM (restricted to lose weight from 21 to 17 kg then 70% ad libitum to 30 kg). Weights of visceral organs and mesenteric and kidney fat showed dramatic responses to alteration of feed allowances. After recovery from 20% live weight loss, weight of liver equaled or exceeded that of both the ad libitum and 70% refed lambs. Mesenteric and kidney fat did not. Refeeding was accompanied by an increase in water (P less than .05) and a decrease in fat (P less than .01) of both carcass and noncarcass components. These results indicate that weight loss of lambs incurred during feed shortage was largely in internal organ weights, and that these lambs can recover these losses during realimentation and undergo compensatory growth with better feed efficiency and lean carcasses.

Effects of undernutrition and refeeding on weights of body parts and chemical components of growing Moroccan lambs.

Forty-four intact, male lambs (20 Timahdit and 24 D'man) were used to assess the effects of 22% (from approximately 25 to approximately 20 kg) and 31% (from approximately 25 to approximately 17 kg) live weight loss and the subsequent refeeding to initial BW on changes in body components. Body composition was determined using a serial slaughter technique at 17, 20, and 25 kg live weight during normal growth, weight loss, and refeeding phases. Reduction in live weight from 25 to 20 kg was associated with greater loss of visceral organs (30%) and internal fat (75%) than carcass loss (19%). Further body weight loss (from 20 to 17 kg) involved carcasses to a greater extent than internal organs. The composition of BW loss consisted of 53% water, 28% fat, and 15% protein. Refeeding was associated with a rapid increase in organ weights and less fat regeneration. Although total internal organs recovered only 90% of their original weight, liver and kidneys regained all their weight. At the same slaughter weight, carcass and noncarcass components of refed lambs were leaner because of lower fat content in these components.