Blood and hair as non-invasive trace element biological indicators in growing rabbits

[EN] The suitability of blood and hair as non-invasive tools to monitor trace element contents was studied in 48 Hyla male growing rabbits. Three diets with increasing organic selenium (Se) addition (0.1, 0.5 and 2.5 mg/kg) were used to induce alterations in the concentrations of trace elements vs. an unsupplemented diet. In blood, a linear decrease in Co (P<0.001), Cu (P<0.001), Mn (P<0.05), Zn (P<0.05), Sb (P<0.001), As (P<0.001), Cr (P<0.001), Mo (P<0.001), Ni (P<0.001) and Cd (P<0.001) concentrations with increasing dietary Se was observed. In hair, a cubic effect of dietary Se on Co (P<0.01), Cu (P<0.05), Mn (P<0.001), Pb (P<0.05), Mo (P<0.05) and Cd (P<0.05) concentrations was found, while As, Cr and Ni concentrations decreased linearly (P<0.01, P<0.01 and P<0.001, respectively) with increasing dietary Se. Selenium was negatively correlated to Sb, As, Cr, Mo, Ni and Cd, (P<0.001) in blood, and to As (P<0.05), Cr, Ni (P<0.01) and Pb (P<0.05) in hair. The contents of Se, As, Cr and Ni in blood were highly correlated (P<0.001) to those in hair. Blood appeared to be more sensitive than hair in detecting small changes in the trace element profile in rabbits, as was indicated by the discriminant analysis. In conclusion, blood and hair can be suitable biological indicators of essential, toxic and potentially toxic trace element status in rabbits, particularly when used complementarily.The authors are grateful to NUEVO S.A. (N Artaki, Euboia, Greece) for providing Sel-Plex®. This research has not received any specific funding.Papadomichelakis, G.; Pappas, AC.; Zoidis, E.; Danezis, G.; Georgiou, KA.; Fegeros, K. (2019). Blood and hair as non-invasive trace element biological indicators in growing rabbits. World Rabbit Science. 27(1):21-30. https://doi.org/10.4995/wrs.2019.10654SWORD2130271Barbosa F.J., Tanus-Santos J.E., Gerlach R.F., Parsons P.J. 2005. A critical review of biomarkers used for monitoring human exposure to lead: advantages, limitations, and future needs. Environ. Health Persp., 113: 1669-1674. https://doi.org/10.1289/ehp.7917Bryan C.E., Christopher S.J., Balmer B.C., Wells R.S. 2007. Establishing baseline levels of trace elements in blood and skin of bottlenose dolphins in Sarasota Bay, Florida: implications for non-invasive monitoring. Sci. Total Environ., 388: 325-342. https://doi.org/10.1016/j.scitotenv.2007.07.046Čobanová K., Chrastinová Ľ., Chrenková M., Polačiková M., Formelová Z., Ivanišinová O., Ryzner M., Grešáková Ľ. 2018. The effect of different dietary zinc sources on mineral deposition and antioxidant indices inrabbit tissues World Rabbit Sci., 26: 241-248. https://doi.org/10.4995/wrs.2018.9206de Blas C., Mateos G.G. 2010. Feed formulation. In 'The Nutrition of the Rabbit (2nd ed.)'. C de Blas, J. Wiseman (Eds.) 222-231. CAB International: Wallingford, UK. https://doi.org/10.1079/9781845936693.0222De Temmerman L., Vanongeval L., Boon W., Hoenig M., Geypens M. 2003. Heavy metal content of arable soils in northern Belgium. Water Air Soil Poll., 148: 61-76. https://doi.org/10.1023/A:1025498629671FEDNA (2003). Fundación Española para el Desarrollo de la Nutrición Animal. In C. De Blas, G. G. Mateos, & P. G. Rebollar (Eds.), Tablas FEDNA de composición y valor nutritivo de alimentos para la fabricación de piensos (2nd ed.). Madrid, Spain: FEDNA.Georgiou C.A., Koupparis M.A. 1990. Automated flow injection spectrophotometric determination of para- and metasubstituted phenols of pharmaceutical interest based on their oxidative condensation with 1-nitroso-2-naphthol. Analyst, 115: 309-313. https://doi.org/10.1039/an9901500309Georgiou C.A., Danezis G.P. 2015. Elemental and isotopic mass spectrometry. In 'Advanced Mass Spectrometry for Food, Comprehensive Analytical Chemistry'. (Ed. Y Pico) 131-243. Elsevier: Amsterdam. https://doi.org/10.1016/B978-0-444-63340-8.00003-0Glynn A.W., Ilback N.G., Brabencova D., Carlsson L., Enqvist E.C., Netzel E., Oskarsson A. 1993. Influence of sodium selenite on 203Hg absorption, distribution and elimination in male mice exposed to methyl 203Hg. Biol. Trace Elem. Res., 39: 91-107. https://doi.org/10.1007/BF02783813Gulson B.L., Mizon K.J., Korsch M.J., Howarth D., Phillips A., Hall J. 1996. Impact on blood lead in children and adults following relocation from their source of exposure and contribution of skeletal tissue to blood lead. B. Environ. Contam. Tox., 56: 543-550. https://doi.org/10.1007/s001289900078Hasan M.Y., Kosanovic M., Fahim M.A., Adem A., Petroianu G. 2004. Trace metal profiles in hair samples from children in urban and rural region of the United Arab Emirates. Vet. Hum. Toxicol., 46: 119-121.He K. 2011. Trace elements in nails as biomarkers in clinical research. Eur. J. Clin. Invest., 41: 98-102.https://doi.org/10.1111/j.1365-2362.2010.02373.xKäkelä R., Käkelä A., Hyvärinen H. 1999. Effects of nickel chloride on reproduction of the rat and possible antagonistic role of selenium. Comp. Biochem. Physiol. C, 123: 27-37.https://doi.org/10.1016/S0742-8413(99)00006-7Kan C.A., Meijer G.A.L. 2007. The risk of contamination of food with toxic substances present in animal feed. Anim. Feed Sci. Technol., 133: 84-108. https://doi.org/10.1016/j.anifeedsci.2006.08.005Keil D.E., Berger-Ritchie J., McMillin G.A. 2011. Testing for toxic elements: a focus on arsenic, cadmium, lead, and mercury. Labmedicine, 42: 735-742. https://doi.org/10.1309/LMYKGU05BEPE7IAWKlotz L.O., Kröncke K.D., Buchczyk D.P., Sies H. 2003. Role of copper, zinc, selenium and tellurium in the cellular defense against oxidative and nitrosative stress. J. Nutr., 133: 1448-1451. https://doi.org/10.1093/jn/133.5.1448SLevander O.A. 1977. Metabolic interrelationships between arsenic and selenium. Environ. Health Persp., 19: 159-164. https://doi.org/10.1289/ehp.7719159López-Alonso M.L., Benedito J.L., Miranda M., Castillo C., Hernández J., Shore R.F. 2002. Cattle as biomonitors of soil arsenic, copper and zinc concentrations in Galicia (NW Spain). Arch. Environ. Contam. Toxicol., 43: 103-108. https://doi.org/10.1007/s00244-002-1168-5McDowell L.R. 2003. 'Minerals in animal and human nutrition (2nd ed)'. (Elsevier Science: Amsterdam).Milošković A., Simić V. 2015. Arsenic and other trace elements in five edible fish species in relation to fish size and weight and potential health risks for human consumption. Pol. J. Environ. Stud., 24: 199-206. https://doi.org/10.15244/pjoes/24929Miranda M., López-Alonso M., Castillo C., Hernández J., Benedito J.L. 2005. Effects of moderate pollution on toxic and trace metal levels in calves from a polluted area of northern Spain. Environ. Int., 31: 543-548.https://doi.org/10.1016/j.envint.2004.09.025Ohta H., Seki Y., Yoshikawa H. 1995. Interactive effects of selenium on chronic cadmium toxicity in rats. ACES Bulletin, 8: 97-104. Othman A.I., El Missiry M.A. 1998. Role of selenium against lead toxicity in male rats. J. Biochem. Mol. Toxic., 12: 345-349. https://doi.org/10.1002/(SICI)1099-0461(1998)12:6%3C345::AID-JBT4%3E3.0.CO;2-VPapadomichelakis G., Zoidis E., Pappas A.C., Mountzouris K.C., Fegeros K. 2017. Effects of increasing dietary organic selenium levels on meat fatty acid composition and oxidative stability in growing rabbits. Meat Sci., 131: 132-138. https://doi.org/10.1016/j.meatsci.2017.05.006Papadomichelakis G., Zoidis E., Pappas A.C., Danezis G., Georgiou C.A., Fegeros K. 2018. Dietary organic selenium addition and accumulation of toxic and essential trace elements in liver and meat of growing rabbits. Meat Sci., 145: 383-388. https://doi.org/10.1016/j.meatsci.2018.07.022Pappas A.C., Zoidis E., Georgiou C.A., Demiris N., Surai P.F., Fegeros K. 2011. Influence of organic selenium supplementation on the accumulation of toxic and essential trace elements involved in the antioxidant systemof chicken. Food Addit. Contam. Part A, 28: 446-454. https://doi.org/10.1080/19440049.2010.549152Park D.U., Kim D.S., Yu S.D., Lee K.M., Ryu S.H., Kim S.G. et al. 2014. Blood levels of cadmium and lead in residents near abandoned metal mine areas in Korea. Environ. Monit. Assess., 186: 5209-5220. https://doi.org/10.1007/s10661-014-3770-1Patra R.C., Swarup D., Naresh R., Kumar P., Nandi D., Shekhar P., Roy S., Ali S.L. 2007. Tail hair as an indicator of environmental exposure of cows to lead and cadmium in different industrial areas. Ecotoxicol. Environ. Saf., 66: 127-131. https://doi.org/10.1016/j.ecoenv.2006.01.005Paukert J., Obrusnik I. 1986. The hair of the common hare (Lepus europaeus Pall.) and of the common vole (Microtus arvalis Pall.) as indicator of the environmental pollution. J. Hyg. Epidem. Microb. Imm., 30: 27-32.Paulsson K., Lundbergh K. 1989. The selenium method for treatment of lakes for elevated levels of mercury in fish. Sci. Total Environ., 87-88: 495-507. https://doi.org/10.1016/0048-9697(89)90256-8Perrone L., Moro R., Caroli M., Universit S., Fisiche S., Federico N. 1996. Trace elements in hair of healthy children sampled by age and sex. Biol. Trace Elem. Res., 51: 71-76. https://doi.org/10.1007/BF02790149Raab A., Hansen H.R., Zhuang L.Y., Feldmenn J. 2002. Arsenic accumulation and speciation analysis in wool from sheep exposed to arsenosugars. Talanta, 58: 167-176.https://doi.org/10.1016/S0039-9140(02)00257-6Reis L.S.L.S., Pardo P.E., Camargo A., Oba E. 2010. Mineral element and heavy metal poisoning in animals. Int. J. Med. Med. Sci., 1: 560-579.Rogowska K.A., Monkiewicz J., Grosicki A. 2009. Lead, cadmium, arsenic, copper, and zinc contents in the hair of cattle living in the area contaminated by a copper smelter in 2006-2008. B. Vet. I. Pulawy, 53: 703-706.Samanta G., Sharma R., Roychowdhury T., Chakraborti D. 2004. Arsenic and other elements in hair, nails, and skinscales of arsenic victims in West Bengal, India. Sci. Total Environ., 326: 33-47. https://doi.org/10.1016/j.scitotenv.2003.12.006Sanna E., Liguori A., Palmas L., Sor M.R., Floris G. 2003. Blood and hair lead levels in boys and girls living in two Sardinian towns at different risks of lead pollution. Ecotoxicol. Environ. Saf., 55: 293-299. https://doi.org/10.1016/S0147-6513(02)00072-6Sarmani S. 1987. A study of trace elements concentrations in human hair of some local population in Malaysia. J. Radioanal. Nucl. Chem., 110: 627-632. https://doi.org/10.1007/BF02035551Shanker K., Mishra S., Srivastava S., Srivastava R., Dass S., Prakash S., Srivastava M.M. 1996. Study of mercuryselenium (Hg-Se) interactions and their impact on Hg uptake by the radish (Raphanus sativus) plant. Food Chem. Toxic., 34: 883-886.https://doi.org/10.1016/S0278-6915(96)00047-6Shen S., Li X.F., Cullen W.R., Weinfeld M., Le X.C. 2013. Arsenic binding to proteins. Chem. Rev., 113: 7769-7792. https://doi.org/10.1021/cr300015cSoudani N., Amara I.B., Sefi M., Boudawara T., Zeghal N. 2011. Effects of selenium on chromium (VI)-induced hepatotoxicity in adult rats. Exp. Toxicol. Pathol., 63: 541-548. https://doi.org/10.1016/j.etp.2010.04.005Templeton G.F. 2011. A two-step approach for transforming continuous variables to normal: implications and recommendations for IS research. Commun. Assoc. Inf. Syst., 28: 41-58. https://doi.org/10.17705/1CAIS.02804Underwood E.J., Suttle N.F. 1999. 'The mineral nutrition of livestock (3rd ed.)'. CAB International: Wallingford, UK. 343-373. https://doi.org/10.1079/9780851991283.0000Valko M., Morris H., Cronin M.T.D. 2005. Metals, toxicity and oxidative stress. Curr. Med. Chem., 12: 1161-1208. https://doi.org/10.2174/0929867053764635Waegeneers N., Pizzolon J.C., Hoenig M., De Temmerman L. 2009. Accumulation of trace elements in cattle from rural and industrial areas in Belgium. Food Addit. Contam. A, 26: 326-332. https://doi.org/10.1080/02652030802429096Wangher P.D. 2001. Selenium and the brain: a review. Nutr. Neurosci., 4: 81-97. https://doi.org/10.1080/1028415X.2001.11747353Xing R., Li Y., Zhang B., Li H., Liao X. 2017. Indicative and complementary effects of human biological indicators for heavy metal exposure assessment. Environ. Geochem. Hlth., 39: 1031-1043. https://doi.org/10.1007/s10653-016-9870-9Zoidis E., Pappas A.C., Georgiou C.A., Komaitis Ε., Feggeros K. 2010. Selenium affects the expression of GPx4 and catalase in the liver of chicken. Comp. Biochem. Physiol. B, 155: 294-300. https://doi.org/10.1016/j.cbpb.2009.11.017Żukowska J., Biziuk M. 2008. Methodological evaluation of method for dietary heavy metal intake. J. Food Sci., 73: 21-29. https://doi.org/10.1111/j.1750-3841.2007.00648.

Effect of Clarithromycin in Patients with Sepsis and Ventilator-Associated Pneumonia

Background. Because clarithromycin provided beneficiary nonantibiotic effects in experimental studies, its efficacy was tested in patients with sepsis and ventilator-associated pneumonia (VAP). Methods. Two hundred patients with sepsis and VAP were enrolled in a double-blind, randomized, multicenter trial from June 2004 until November 2005. Clarithromycin (1 g) was administered intravenously once daily for 3 consecutive days in 100 patients; another 100 patients were treated with placebo. Main outcomes were resolution of VAP, duration of mechanical ventilation, and sepsis-related mortality within 28 days. Results. The groups were well matched with regard to demographic characteristics, disease severity, pathogens, and adequacy of the administered antimicrobials. Analysis comprising 141 patients who survived revealed that the median time for resolution of VAP was 15.5 days and 10.0 days among placebo- and clarithromycin-treated patients, respectively (P=.011); median times for weaning from mechanical ventilation were 22.5 days and 16.0 days, respectively (P=.049). Analysis comprising all enrolled patients showed a more rapid decrease of the clinical pulmonary infection score and a delay for advent of multiple organ dysfunction in clarithromycin-treated patients, compared with those of placebo-treated patients (P=.047). Among the 45 patients who died of sepsis, time to death was significantly prolonged in clarithromycin-treated compared with placebo-treated patients (P=.004). Serious adverse events were observed in 0% and 3% of placebo- and clarithromycin-treated patients, respectively (P=.25). Conclusions. Clarithromycin accelerated the resolution of VAP and weaning from mechanical ventilation in surviving patients and delayed death in those who died of sepsis. The mortality rate at day 28 was not altered. Results are encouraging and render new perspectives on the management of sepsis and VA

Effects of Selenium and Cadmium on Breast Muscle Fatty-Acid Composition and Gene Expression of Liver Antioxidant Proteins in Broilers

The present work was part of a project intended to evaluate whether organic selenium (Se) has the potential to protect against toxic effects exerted by cadmium (Cd). For this reason, 300 as-hatched, one-day-old broiler chickens were randomly allocated in four dietary treatments with five replicate pens per treatment. Chickens in T1 treatment, were offered a diet supplemented with 0.3 ppm Se (as Se-yeast), without added Cd; in T2 treatment, they were offered a diet with 0.3 ppm Se and 10 ppm Cd; in T3 treatment, they were offered a diet with 0.3 ppm Se and 100 ppm Cd; in T4 treatment, chickens were offered a diet supplemented with 3 ppm Se and 100 ppm Cd. Cadmium was added to the diets in T2, T3, and T4 as CdCl2. On the fourth and sixth weeks, liver and breast samples were obtained from two broilers per replicate pen. Relative gene expression levels of catalase (CAT), superoxide dismutase 1 (SOD1) and 2 (SOD2), methionine sulfoxide reductase A (MSRA) and B3 (MSRB3), iodothyronine deiodinase 1 (DIO1), 2 (DIO2), and 3 (DIO3), glutathione peroxidase 1 (GPX1) and 4 (GPX4), thioredoxin reductase 1 (TXNRD1) and 3 (TXNRD3), and metallothionein 3 (MT3) were analyzed by real-time quantitative PCR in liver, whereas the fatty-acid (FA) profile of breast muscle was determined by gas chromatography. Broilers supplemented with 0.3 ppm Se could tolerate low levels of Cd present in the diets, as there were no significant changes in the breast muscle FA profile, whereas excess Cd led to decreased polyunsaturated fatty acids (PUFAs), and in particular n-6 PUFA. Furthermore, treatments mainly affected the messenger RNA (mRNA) expression of SOD2, TXNRD3, and MT3, while age affected CAT, MSRB3, DIO2, DIO3, GPX4, TXNRD1, and MT3. In conclusion, dietary Se may help against the negative effects of Cd, but cannot be effective when Cd is present at excessive amounts in the diet

Screening for resistant gram-negative microorganisms to guide empiric therapy of subsequent infection

To define the potential of resistant gram-negative colonization surveillance to predict etiology of subsequent infection and improve adequacy of empiric antimicrobial treatment. Retrospective cohort study. A mixed medical-surgical six-bed intensive care unit (ICU), from November 2003 to December 2006. All patients having at least one episode of ventilator-associated pneumonia (VAP) or bloodstream infection (BSI) caused by a resistant gram-negative pathogen during the study period. Colonization surveillance of the respiratory tract and gastrointestinal tract was systematically performed in all ICU patients. Tracheal aspirates were obtained twice weekly and rectal swabs once weekly. Both tracheal and rectal samples were cultured in antibiotic-enriched media (containing ceftazidime, ciprofloxacin, imipenem or piperacillin/tazobactam), to focus on resistant gram-negative pathogen isolation. Colonization concordance between resistant, gram-negative pathogens of infectious episodes and previous, recent (&lt;= 7 days) colonization of the respiratory and gastrointestinal tract was determined, based on species identity and antimicrobial susceptibility. Concordance was 82% in VAP and 86% in BSI cases and was further confirmed by molecular testing of 15 randomly selected cases by REP-PCR. Previous colonization had high sensitivity and specificity in VAP, but was less specific in BSI cases. Knowledge of previous colonization improved the rate of adequate empiric antimicrobial treatment (91 vs. 40% in VAP and 86 vs. 50% in BSI cases, P &lt; 0.05). Colonization surveillance for resistant gram-negative microorganisms is predictive of subsequent infection etiology and can improve empiric antimicrobial treatment adequacy in a critical care setting

Dietary Orange Pulp and Organic Selenium Effects on Growth Performance, Meat Quality, Fatty Acid Profile, and Oxidative Stability Parameters of Broiler Chickens

In this study, orange pulp (OP) and/or organic Se were fed to broilers in order to investigate their effects on the performance, behavior, breast meat quality, and oxidative stability. A total of 240 chicks were allocated to four groups: a control group; an OP group, fed with OP at 50 g/kg of diet; a Se group, fed with organic Se at 0.15 ppm; and an OP + Se group, fed with OP and organic Se at 50 g/kg and 0.15 ppm, respectively. The selenium and OP + Se groups showed improved meat oxidative stability during frozen storage from 90 to 210 days (p p > 0.05), apart from a reduction in the meat pH and the dressing percentage in the OP-supplemented groups (p p p > 0.05). The dietary supplementation of broiler chickens with the citrus industry byproduct orange pulp at 50 g/kg, along with organic Se at 0.15 ppm, beneficially improves the meat oxidative stability and the meat nutritional value, with no negative side effects on the performance or the meat quality

Clinical experience of serious infections caused by Enterobacteriaceae producing VIM-1 metallo-beta-lactamase in a Greek university hospital

Background. The dissemination of acquired metallo-beta-lactamases (MBLs) in members of the family Enterobacteriaceae is regarded as an emerging clinical threat. The clinical characteristics and outcomes of 17 cases of infection due to MBL-producing isolates were analyzed. Methods. During a 3-year period, medical records for all patients with confirmed infection due to an MBL-producing strain belonging to the Enterobacteriaceae family were retrospectively analyzed. We screened for MBL production with the imipenem-ethylenediaminetetraacetic acid disk synergy test, and results were confirmed by polymerase chain reaction. Genetic relatedness between isolates was evaluated by repetitive extragenic palindromic polymerase chain reaction. Results. Fourteen cases of bacteremia and 3 cases of ventilator-associated pneumonia due to an MBL-producing isolate were studied. Most of the patients had previously been colonized with an MBL-producing organism, and almost 60% had been exposed to carbapenems before infection. The isolated pathogens (Klebsiella pneumoniae, 14 cases; and Klebsiella oxytoca, Enterobacter cloacae, and Enterobacter aerogenes, 1 case each) exhibited variable minimum inhibitor concentrations of carbapenems (1 to &gt; 32 mu g/mL) and resistance to most other beta-lactams. Tigecycline was active against all isolates, whereas colistin and gentamicin were active against 88% of them. Molecular studies confirmed the presence of a gene belonging to bla(VIM-1) cluster in all isolates. Among the 12 K. pneumoniae isolates, which were subjected to molecular typing, 11 distinct clones were identified. Five cases (similar to 30%) occurred in patients who were already receiving carbapenem-containing treatment, and carbapenem treatment was considered to have failed. Twelve cases were treated with a colistin-containing regimen. The attributable mortality rate was 18.8%. Conclusions. MBL-producing Enterobacteriaceae can cause severe, often fatal infection in severely ill patients, irrespective of the MIC of carbapenems. Colonization with an MBL-producer is a preceding event, highlighting the importance of surveillance. Both infection control practices and antibiotic policies should be intensified to contain the spread of these problematic bacteria

Dietary Orange Pulp and Organic Selenium Effects on Growth Performance, Meat Quality, Fatty Acid Profile, and Oxidative Stability Parameters of Broiler Chickens

In this study, orange pulp (OP) and/or organic Se were fed to broilers in order to investigate their effects on the performance, behavior, breast meat quality, and oxidative stability. A total of 240 chicks were allocated to four groups: a control group; an OP group, fed with OP at 50 g/kg of diet; a Se group, fed with organic Se at 0.15 ppm; and an OP + Se group, fed with OP and organic Se at 50 g/kg and 0.15 ppm, respectively. The selenium and OP + Se groups showed improved meat oxidative stability during frozen storage from 90 to 210 days (p &lt; 0.05), whereas the performance and meat quality were unaffected by the dietary treatments (p &gt; 0.05), apart from a reduction in the meat pH and the dressing percentage in the OP-supplemented groups (p &lt; 0.05). A synergistic action between OP and Se was observed for the meat oxidative stability. The polyunsaturated fatty acid (FA) and &alpha;-linolenic acid (ALA) contents in the breast meat lipid fractions were increased in the OP groups (p &lt; 0.05). Dietary intervention did not affect the feeding or drinking behaviors of the broilers (p &gt; 0.05). The dietary supplementation of broiler chickens with the citrus industry byproduct orange pulp at 50 g/kg, along with organic Se at 0.15 ppm, beneficially improves the meat oxidative stability and the meat nutritional value, with no negative side effects on the performance or the meat quality

Colistin-resistant isolates of Klebsiella pneumoniae emerging in intensive care unit patients: first report of a multiclonal cluster

Objectives: Infections due to multidrug-resistant (MDR) Gram-negative pathogens in the ICU have prompted the use of colistin, an antibiotic forgotten for decades. The aim of this retrospective observational study was to record and present the emergence of colistin-resistant Klebsiella pneumoniae (CRKB) in a Greek ICU. Methods: In a new university tertiary hospital, the first patients admitted in the ICU were already colonized or infected with MDR pathogens, and this led to frequent colistin use as part of empirical or microbiological ly documented therapy. Colistin resistance was defined as MIC &gt;4 mg/L by the Etest method. All CRKB isolated in surveillance cultures or clinical specimens in the ICU during the period 2004-5 were recorded along with patients’ characteristics. Results: Eighteen CRKB were isolated from 13 patients over a 16 month period, representing either colonizing or infective isolates. Patients’ mean age was 70 years, with a mean APACHE 11 score at admission of 22. They all had a long hospitalization (median 69 days) and a long administration of colistin (median 27 days). Colistin-resistant isolates were implicated as pathogens in two bacteraemias, a ventilator-associated pneumonia and two soft tissue infections. Repetitive extragenic palindromic PCR identified six distinct clones, and horizontal transmission was also documented. Conclusions: Selective pressure due to extensive or inadequate colistin use may lead to the emergence of colistin resistance among K. pneumoniae isolates, jeopardizing treatment options in the ICU, potentially increasing morbidity and mortality of critically ill patients and necessitating prudent use of colistin

Inhaled activated protein C attenuates lung injury induced by aerosolized endotoxin in mice

The serine protease activated protein C (APC) possesses prominent anticoagulant and anti-inflammatory actions. In this study, we investigated the effect of inhaled recombinant human (rh) APC in a murine lung injury model. Animals inhaled 10 mg of Pseudomonas lipopolysaccharide (LPS) in 3 mL normal saline (NS); 3 0 min prior to LPS, mice were pretreated with inhaled rhAPC (4 mg/3 mL NS; APC + LPS group) or NS (LPS group), A control animal group inhaled vehicle (NS) twice. 24 h later, total cells and cell-types, protein content, and the cytokines tumor necrosis factor-a, interleukin (IL)-6, macrophage inflammatory protein-la, and mouse keratinocyte-derived chemokine (a homolog of human IL-8) were estimated in bronchoalveolar lavage fluid (BALF). Lung pathology given as total histology score (THS), wet/dry lung weight ratios, and lung vascular cell adhesion molecule (VCAM)-1 expression were additionally assessed. rhAPC inhalation attenuated the aerosolized LPS-induced increases of. total cells, nentrophils and macrophages in BALF, lung tissue VCAM-1 protein levels, and THS. Total protein levels and cytokines in BALF, and wet/dry weight ratios were increased in the LPS group, but rhAPC pretreatment did not significantly alter the LPS-induced responses. In conclusion, in this murine septic model of lung injury, inhaled rhAPC appears to attenuate lung inflammation, without reversing the observed increases in lung permeability and BALF cytokines. This effect may be associated with leukocyte trafficking modifications, related, at least in part, to VCAM-1 reduction. (c) 2006 Elsevier Inc. All rights reserved