Prolonging culture to 15 days improves bacterial detection in bone and joint infections.

Since the optimal incubation period of cultures for diagnosis of bone and joint infections is still a matter of debate, the present study aimed to evaluate the effects of different incubation periods (5 and 15 days) on microbial isolation. Samples from 387 patients with bone and joint infections (including prosthetic ones) were analyzed from March 2012 to February 2014. In 197 patients (51 %) growth was obtained within 48 hrs, while in 124 (32 %) and 66 (17 %) patients cultures yielded positive results within and after 5 days of incubation, respectively. Of 449 microorganisms isolated, 247 grew within 48 hrs, 131 within the first 5 days of incubation while 71 were isolated after 5 days. Staphylococcus aureus was the most frequently isolated pathogen within 48 hrs, while Propionibacteria were prevalently isolated after 5 days of incubation. Interestingly, about 25 % of microorganisms isolated after 5 days of incubation were coagulase-negative staphylococci. Extending incubation period of broth cultures improves isolation rates of pathogens involved in bone and joint infections thus improving management of these infections.

A comparative study of the use of bioactive glass S53P4 and antibiotic-loaded calcium-based bone substitutes in the treatment of chronic osteomyelitis: a retrospective comparative study.

The treatment of chronic osteomyelitis often includes surgical debridement and filling the resultant void with antibiotic-loaded polymethylmethacrylate cement, bone grafts or bone substitutes. Recently, the use of bioactive glass to treat bone defects in infections has been reported in a limited series of patients. However, no direct comparison between this biomaterial and antibiotic-loaded bone substitute has been performed. In this retrospective study, we compared the safety and efficacy of surgical debridement and local application of the bioactive glass S53P4 in a series of 27 patients affected by chronic osteomyelitis of the long bones (Group A) with two other series, treated respectively with an antibiotic-loaded hydroxyapatite and calcium sulphate compound (Group B; n = 27) or a mixture of tricalcium phosphate and an antibiotic-loaded demineralised bone matrix (Group C; n = 22). Systemic antibiotics were also used in all groups. After comparable periods of follow-up, the control of infection was similar in the three groups. In particular, 25 out of 27 (92.6%) patients of Group A, 24 out of 27 (88.9%) in Group B and 19 out of 22 (86.3%) in Group C showed no infection recurrence at means of 21.8 (12 to 36), 22.1 (12 to 36) and 21.5 (12 to 36) months follow-up, respectively, while Group A showed a reduced wound complication rate. Our results show that patients treated with a bioactive glass without local antibiotics achieved similar eradication of infection and less drainage than those treated with two different antibiotic-loaded calcium-based bone substitutes.

Behaviour of perioperative values of haemoglobin, haematocrit and red blood cells in elderly patients undergoing lower limb arthroplasty: a retrospective cohort study on non-transfused patients.

Little is known on how elderly patients recover pre-operative haemoglobin, haematocrit and red blood cell levels after total hip and knee arthroplasties. In this study we aimed to evaluate blood loss and recovery blood levels in relation to gender, type of surgery and preoperative haemoglobin values. We conducted a retrospective cohort study on 187 patients over 65 years of age who underwent total knee or total hip arthroplasty between January 2008 and December 2009. Preoperative blood analysis was carried out within 40 days prior to intervention followed by a 15-day postoperative follow-up. Haemoglobin recovery values in anaemic patients versus healthy patients was also estimated. All tested values decreased significantly during the first 3-5 postoperative days. Haemoglobin levels decreased statistically significantly more in males than in females, while no significant differences were observed for haematocrit and erythrocytes. Recovery of haemoglobin values did not differ significantly between healthy patients and patients with preoperative haemoglobin below 120 g/L. Furthermore, our data showed a higher blood loss in total hip arthroplasty, whilst recovery rates showed to be higher after a total knee arthroplasty procedure. In conclusion, the type of intervention and gender played an important role in blood loss and recovery rates in total joint arthroplasty.

Scientific evidence of three different insecticide-resistant profiles in Triatoma infestans (Hemiptera: Reduviidae) populations from Argentina and Bolivia.

Triatoma infestans (Klug, 1834) (Hemiptera, Reduviidae) is the main vector of Chagas disease in the southern cone South America. Chemical control to the vectors appears to be the best option to reduce the incidence of the disease. However, since 2002, high resistance to insecticides that correlated with field control failures was detected in T. infestans from Argentina and Bolivia. In this paper, we analyzed three T. infestans populations whose pyrethroid-resistance had been recently detected, and we defined at least three resistant profiles according to the toxicological and biochemical characteristics of the studied resistant populations. The resistance profiles were identified as Ti-R1, Ti-R2, and Ti-R3, corresponding to the Argentinean Acambuco, and the Bolivians Entre Ríos and Mataral. Ti-R1 exhibited nymphs and eggs with medium resistance level to deltamethrin (RR = 32.5 and 28.6; respectively). Pyrethroid-esterases played a relevant role in deltamethrin resistance. Ti-R2 exhibited nymphs with high resistance to deltamethrin (RR = 173.8) and low resistance to fipronil (RR = 12.4). Pyrethroid-esterases were involved in resistance. Moreover, eggs showed medium resistance level to deltamethrin (RR = 39.1). Ti-R3 had nymphs with low resistance to deltamethrin (RR = 17.4), and medium resistance to fipronil (RR = 66.8). Pyrethroid-esterases showed increased activity, and eggs possessed low resistance to deltamethrin (RR = 8.4). The characterization of the resistance to pyrethroid in these T. infestans populations from Argentina and Bolivia do not permit the generalization of three forms of resistance profile. So far as we appear to know, the forms of mechanisms and their frequencies reported here are selected independently, so additional sites might well show additional combinations of resistance mechanisms and their frequencies.

Iron status evaluation as a marker of postoperative joint infection: a pilot study.

We evaluated the effect of different inflammatory conditions on iron status and, as a consequence, the possible use of iron markers as indicators of infection in the diagnosis of postoperative prosthetic orthopaedic joint infections. The study population was consisted of 26 patients undergoing revision of total hip or total knee joint arthroplasty and subdivided into three groups according to the cause of prosthesis implant failure: 10 as having had previous infection (Group A), 10 patients were categorized as having infection (Group B); and the remaining 6 (Group C) as not having infection. These patients were assayed for mean corpuscular haemoglobin concentration (MCHC) and serum values of iron (Fe), ferritin (Fer), transferrin (Tf), soluble transferrin receptor (sTfR), and transferrin saturation (sat Tf). Septic patients display statistically significant lower serum iron concentration, higher sTfR and ferritin levels, lower, but not statistically significant, MCHC compared to non septic ones. Little differences were observed for Tf, sat Tf, tibc, TfR index, among the three groups of patients. Our study suggests that iron status parameters, in particular serum iron, ferritin, sTfR and TfR index, could be useful tools for the early detection and the diagnosis of orthopaedic prosthetic joint infections.

Procalcitonin, C-reactive protein, interleukin-6, and soluble intercellular adhesion molecule-1 as markers of postoperative orthopaedic joint prosthesis infections.

There is a universally recognized need to identify new, reliable markers of inflammation that can aid in the rapid diagnosis of orthopaedic joint prosthesis infections (OJP-Is). Since prompt diagnosis is key to timely intervention in the course of infection, different molecules have been studied. In this study, we examined three groups of patients: those with prosthesis infection, those without infection, and a third group with previous infection in whom the infection had been cleared. Four presumed markers of infection were tested: procalcitonin (PCT); C-reactive protein (CRP); interleukin-6 (IL-6); and soluble intercellular adhesion molecule-1 (sICAM-1). The results showed that PCT cannot be considered as a good marker of periprosthetic infection as no statistically significant difference in serum PCT levels emerged between patients with infection and controls or patients without infection. In contrast, both sICAM-1 and CRP may be considered as good markers of infection, as measurement of their levels allowed us to distinguish between patients with and without infection, and between patients with infection and those with previous infection, since marker levels quickly returned to baseline values after clearance of the infection. IL-6 was found to be a good marker for inflammation, as it distinguished between patients with infection and the other groups. In the patients with previous infection, the IL-6 values remained high versus the controls but lower and with a statistically significant difference versus the patients with infection. Further studies are needed to determine the cut-off value of IL-6 between patients with infection and those with previous infection.

New findings of insecticide resistance in Triatoma infestans (Heteroptera: Reduviidae) from the Gran Chaco.

Chagas disease is a major health concern in Latin America, and Triatoma infestans (Klug, 1834) is responsible for the majority of cases of Chagas disease in the continent. After the discovery of deltamethrin highly resistant populations in the neighboring areas to Yacuiba (Bolivia) and Salvador Mazza (Argentina), we studied T. infestans populations north and southward, with the aim of describing the range of the resistant area. In addition, tests were conducted to describe the susceptibility to fipronil in deltamethrin-resistant populations. Tarija populations were highly resistant to deltamethrin, showing that the resistant area is greater than previously reported. Argentinean and Paraguayan populations were susceptible or presented moderate to low levels of deltamethrin resistance. Resistance to fipronil was found in Bolivian populations, although this insecticide was effective against Argentinean populations. This study shows that the Argentinean and Paraguayan populations are currently under successful control with deltamethrin. However, continuous resistance monitoring is necessary. We found that fipronil is a viable option for Argentina, whereas the resistance found in Bolivia implies that new formulations are needed to control T. infestans in Bolivia. Further research is required to find new alternatives of control in those areas that are currently suffering from high infestation rates.

Ovicidal and adulticidal effects of monoterpenoids against permethrin-resistant human head lice, Pediculus humanus capitis.

The improper use of pediculicides containing permethrin has led to the development of resistance. Thus, new alternatives for control are needed. Plant-derived insecticides are attractive alternatives to common chemical insecticides because most of them are environmentally friendly and non-toxic to mammals. The toxic activity of 23 monoterpenoids belonging to several chemical classes was tested against the eggs of permethrin-resistant head lice, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae). Significant differences in ovicidal action were observed among the tested substances. The most effective chemicals were hydrocarbons and ethers, followed by ketones, alcohols, phenols and esters. A linear relationship between egg mortality and knockdown time (KT(50)) on adults by the tested components revealed that most of the components were effective on both egg and adult stages. The monoterpenoids described herein are good candidates as effective pediculicides.

Volatile compounds secreted by Brindley's glands of adult Triatoma infestans: identification and biological activity of previously unidentified compounds.

Volatile emissions of adult male Triatoma infestans were collected on non-polar SPME fibers and analyzed by gas chromatography linked to a mass spectrometer. A complex mixture of 16 short-chain esters and acids were identified. The composition of short-chain aliphatic acids (ethanoic to nonanoic acids) was similar to previously reported results. The most abundant aliphatic acid was 2-methylpropanoic acid, constituting 18% of the total volatile content. Also abundant were the esters 2- and 3-methylbutyl 2-methylpropanoate, which constituted 30% and 22%, respectively, of the total volatile content. A similar pattern of compounds was observed in the volatiles secreted by dissected male Brindley's glands; however, in this case, 2- and 3-methylbutan-1-ol were detected which were not found in live insect volatile emissions. Large variability in volatile composition was also observed among the glands excised from different insects. Electroantennographic (EAG) evaluation of the components of Brindley's gland showed significant responses for 2- and 3-methylbutyl 2-methylpropanoate compared to controls. The mixture of volatiles secreted by excised Brindley's glands and the isolated 2- and 3-methylbutyl 2-methylpropanoate had repellent effects on both male and female T. infestans, possibly associated with a defensive strategy.

Increased monooxygenase activity associated with resistance to permethrin in Pediculus humanus capitis (Anoplura: Pediculidae) from Argentina.

We studied the profile of permethrin resistance in populations of head lice infesting children 6-12 yr old in schools and their homes in and around Buenos Aires, Argentina. Five permethrin-resistant populations with different levels of resistance were collected: Hogar Loyola (HL), Republica de Turquia (RT), Hogar Mitre (HM), Guardia de Honor (GH), and Ricardo Guiraldes (RG). One susceptible population, Bandera Argentina (BA), also was collected. Their level of resistance was evaluated, and results showed resistance ratios of 13 for HL, 16 for RT, 22 for HM, 61 for GH, and 69 for RG. To elucidate the possible involvement of the cytochrome P450 monooxygenase system in conferring permethrin resistance, ethoxycoumarin-O-deethylase (ECOD) activity was measured in abdomens of individual third instars and adults by using a fluorometric assay. The ECOD activity was lower in the susceptible BA population (4.7 ng per louse) than in the resistant ones (13.7 ng per louse for RG, 12.3 ng per louse for GH, 8.6 ng per louse for RT, and 8.2 ng per louse for HL). ECOD activity was significantly correlated with the level of resistance in the field populations (r = 0.97, P = 0.0009), suggesting a role for cytochrome monooxygenase P450 system in permethrin resistance by head louse, Pediculus humanus capitis De Geer.

Prevalence and levels of permethrin resistance in Pediculus humanus capitis De Geer (Anoplura: Pediculidae) from Buenos Aires, Argentina.

Permethrin has been used extensively for control of Pediculus humanus capitis De Geer (Anoplura: Pediculidae) in Argentina since 1990, resulting in the development of resistance to this and other pyrethroids. This resistance was first detected in some field populations in 1997. A survey for resistance in Buenos Aires in 2001 revealed significant resistance levels in lice on children at 24 of 26 (92.3%) schools. When compared with a previously unexposed reference population, resistance ratios (RRs) obtained by exposing the insects to filter papers impregnated with permethrin ranged from 2 to 60 in 10 (39%) of the schools. RRs in the remaining 14 (61%) populations were too great to not be measured with the filter paper method (RR > 88.7). As an alternative, we used topical applications of 0.1 microl of acetone solution of permethrin on the dorsal abdomen of adults and third instars. This topical method, which has not been previously reported for head lice, was capable of quantifying higher levels of resistance. Highly resistant populations had RRs from 162.5 to 655.2. When applied to populations with low and intermediate levels of resistance, results from the filter paper and topical application methods were highly correlated, and RRs from topical application were higher than those from the filter paper method. Results from the combination of the two methods indicated that head lice among Buenos Aries school children are highly resistant to permethrin, and the resistance is widespread.

Toxic effect of aliphatic alcohols against susceptible and permethrin-resistant Pediculus humanus capitis (Anoplura: Pediculidae).

The effectiveness of 1-octanol, 1-nonanol, 1-decanol, 1-undecanol, and 1-dodecanol was evaluated by immersion method against susceptible and permethrin-resistant head lice, Pediculus humanus capitis De Geer, from Buenos Aires, Argentina. All the tested alcohols showed knockdown effect at 10 min and mortality 18 h after treatment. The highest activity was found for the 1-dodecanol (KC50 2.55%, LC50 2.28%) and the lowest for 1-octanol (KC50 8%, LC50 4.46%). The toxicity to the head lice systematically increased with the increase in carbon atoms in the n-aliphatic alcohol moiety, and with the octanol:water coefficient (r2 = 0.94). The pediculicidal activity of 1-dodecanol was not correlated with resistance to permethrin, because no significant difference was observed between toxicity parameters in the susceptible (MAR) and the permethrin-resistant populations which had different resistant levels (RR 5.77 x for E49 population, RR 9.5 x for HL population and RR > 35.3 x for GH population). The pediculicidal effect of aliphatic alcohols demonstrated in this study and the lack of correlation with the permethrin resistance may prove to have a practical value for use in susceptible and permethrin-resistant head lice control.

Resistance to insecticides and effect of synergists on permethrin toxicity in Pediculus capitis (Anoplura: Pediculidae) from Buenos Aires.

Permethrin-resistant colonies of Pediculus capitis (De Geer) from Buenos Aires were used to establish a resistance profile and to examine resistance mechanisms. All permethrin-resistant head lice (resistance ratio from 52.8 to > 88.7) were also resistant to d-phenothrin (resistance ratio from 40.86 to > 48.39) and deltamethrin (resistance ratio from 16.24 to 38.06). No cross-resistance to carbaryl was found in any of the pyrethroid-resistant P. capitis tested. Otherwise, all resistant colonies showed low to high levels of resistance to beta-cypermethrin. This pyrethroid had never been applied as a pediculicide in Argentina; however, the high level of resistance found in these permethrin-resistant colonies (resistance ratio from 9.74 to 50.97) demonstrated that pyrethroid cross-resistance occurred to this novel insecticide. Treatment with piperonyl butoxide (PBO) or triphenylphosphate (TPP) significantly decreased the toxicity of permethrin in the four colonies tested. The esterase inhibitor TPP produced lower enhancement of toxicity than the multifunction oxidase inhibitor PBO in the colonies having the highest resistance levels. Results presented here concerning the cross-resistance profile and synergism by enzyme inhibitors in permethrin-resistant head lice demonstrated that enhanced metabolism was involved in the pyrethroid resistance. However, the substantial degree of resistance that remained after synergism suggested the presence of another resistance mechanism. Cross-resistance to pyrethroid and susceptibility to the carbamate carbaryl suggested a common action mechanism.

Insecticide resistance in Brazilian Triatoma infestans and Venezuelan Rhodnius prolixus.

Resistance levels to insecticides used in control of Chagas Disease vectors were assessed in two species of bugs (Hemiptera: Reduviidae): Triatoma infestans (Klug) from Brazil and Rhodnius prolixus Stål from Venezuela. The resistance ratios, compared to susceptible laboratory strains, were determined by topical application bioassays. The T. infestans PA strain exhibited resistance ratios of 7x to deltamethrin, 3.6x to beta-cyfluthrin and 3.3x to cypermethrin, but was susceptible to beta-cypermethrin and lambda-cyhalothrin. Rhodnius prolixus CA strain showed resistance to all the pyrethroids evaluated, the resistance ratios ranging between 4.5x to lambda-cyhalothrin and 12.4x to cypermethrin. Deltamethrin resistance in both strains was decreased by piperonyl butoxide, suggesting oxidative metabolism as cause of resistance.

Laboratory studies of susceptibility and resistance to insecticides in Pediculus capitis (Anoplura; Pediculidae).

The susceptibility of local head lice to permethrin, sumithrin, deltamethrin, and carbaryl was determined by laboratory bioassays in field-collected colonies. Head lice collected from the infested heads of children 6-12 yr old were tested within 3 h of collection. The longest survival of control insects in the laboratory was obtained by keeping them in the dark at 18 degrees C and 70-80% RH. The base line susceptibility data obtained for insects collected from children not treated for lice, the reference colony, showed that deltamethrin caused the highest mortality of the insecticides tested (LC50, 0.06%). Permethrin, sumithrin, and carbaryl showed no significant difference in mortality (superposition of confidence intervals), being 10 times lower than that caused by deltamethrin. All field-collected lice required a higher LC50 of permethrin than the reference colony. Resistance levels varied from 3 to > 100 for colonies that were taken from children treated with anti-lice products. Lice colonies with permethrin resistance showed resistance to sumithrin and deltamethrin, but resistance was not observed to the carbamate carbaryl.

Reduction of Triatoma infestans (Hemiptera:Reduviidae) experimental populations exposed to feeding inhibitors.

We found that feeding behavior of Triatoma infestans (Klug) 5th-instar nymphs was blocked by topical application of the cis-isomers of methyl esters of N-substituted maleamic acids, synthesized in our laboratory. The antifeeding effect of these compounds was determined by exposing 5th-instar nymphs to impregnated filter papers. The greatest antifeeding effect was observed for the shortest alkyl chain of cis-N-substituted methyl maleamate (EC50: 2.47 micrograms/cm2 when the substituent was ethyl and 65.1 micrograms/cm2 when it was octil). The continuous exposure of a T. infestans population to ethyl, buthyl, heptil, or octil maleamates at an initial concentration of 0.1 mg/cm2 produced significant (P < 0.05) decrease in the development of the exposed populations ranging from 220 to 400 d for the different maleamates assayed. These results suggest the antifeeding maleamates as a specific alternative method of T. infestans control.

Feeding and mating deterrency by sulfhydryl reagents in Triatoma infestans.

Topical application of different sulfhydryl (SH) reagents in acetone solution resulted in a deterrence of the feeding activity of Triatoma infestans nymph V. The ED50 for topical application of N-ethylmaleimide (NEM), the more active compound assayed, was 8.9 micrograms/insect. This effect showed a dose-dependent reversibility. We also observed feeding deterrency when a gauze cloth impregnated with NEM was located between the food source and the nymphs. After topical NEM treatment, mating deterrency was assessed by counting spermatophores dropped and eggs laid. When a population of adult insects, nymphal stages and eggs was reared in a cage containing filter papers treated with 0.4 mg/cm2 of NEM, a significant decrease of population density was seen after 200 days of exposure. Weekly replacement of filter papers impregnated with 0.13 mg/cm2 NEM for a period of one year enabled population control to be maintained. Antifeeding and antimating effects of NEM in Triatoma infestans and the resulting population control could be attributed to a chemoreceptor blockage produced by SH reagents.