Efficacy of antibiotic combinations in an experimental sepsis model with Pseudomonas aeruginosa.

This study aimed to compare the efficacy of fosfomycin, colistin, tobramycin and their dual combinations in an experimental sepsis model. After sepsis was established with a Pseudomonas aeruginosa isolate (P1), antibiotic-administered rats were divided into six groups: Fosfomycin, tobramycin, colistin and their dual combinations were administered by the intravenous or intraperitoneal route to the groups. The brain, heart, lung, liver, spleen and kidney tissues of rats were cultured to investigate bacterial translocation caused by P1. Given the antibiotics and their combinations, bacterial colony counts in liver tissues were decreased in colistin alone and colistin plus tobramycin groups compared with control group, but there were no significant differences. In addition, a non-statistical decrease was found in the spleen tissues of rats in the colistin plus tobramycin group. There was a > 2 log10 CFU/ml decrease in the number of bacterial colonies in the kidney tissues of the rats in the fosfomycin group alone, but the decrease was not statistically significant. However, there was an increase in the number of bacterial colonies in the spleen and kidney samples in the group treated with colistin as monotherapy compared to the control group. The number of bacterial colonies in the spleen samples in fosfomycin plus tobramycin groups increased compared to the control group. Bacterial colony numbers in all tissue samples in the fosfomycin plus colistin group were found to be close to those in the control group. Colistin plus tobramycin combinations are effective against P. aeruginosa in experimental sepsis, and clinical success may be achieved. New in vivo studies demonstrating the ability of P. aeruginosa to biofilm formation in tissues other than the lung are warranted in future.

Relationship between Blastocystis spp. with Accompanying Food Hypersensitivity and Chronic Spontaneous Urticaria.

INTRODUCTION: The relationship between Blastocystis spp. and chronic spontaneous urticaria (CSU) is unclear. The aim of this study was to evaluate the role of this parasitic infection on CSU and to search for risky groups in CSU patients with this parasite. METHODS: Seventy adult CSU patients with Blastocystis spp. in their stool samples forming Group A and 70 CSU patients without any parasite as Group B were prospectively compared regarding urticaria activity score-7 (UAS7), medication scores (MS), and laboratory parameters. All patients received CSU treatment, and additionally, those in group A received an antiparasitic antibiotic. Eight months later, the same parameters were compared between the ones in remission (group A1) and those still having CSU symptoms (group A2) in group A. RESULTS: UAS7 and MS were lower in group A than in group B (p: 0.007, p < 0.001) 8 months later, while the initial scores were similar. The presence of food hypersensitivity reactions (FHRs) was higher in group A than in group B (p < 0.001) and was detected as a significant risk factor in the presence of Blastocystis spp. infection (p: 0.002, OR [CI] = 0.151 [0.045-0.502]). In group A, UAS7, MS, serum total IgE levels, and blood eosinophil counts decreased 8 months later (p < 0.001, p < 0.001, p: 0.003, p: 0.004, respectively). Additionally, total IgE levels and eosinophil counts decreased in group A1 (p: 0.033, p: 0.002) while they did not change in group A2. DISCUSSION/CONCLUSION: The eradication of Blastocystis spp. can improve the disease activity in CSU and the presence of FHRs seems to be risky in CSU patients with Blastocystis spp.

Successful Treatment of Acanthamoeba Meningoencephalitis in an Immunocompetent Child.

Acanthamoeba meningoencephalitis is a very rare entity with high mortality rate. Its diagnosis is usually delated, and the optimal approach for its treatment is uncertain. In presented case, early diagnosis was made with direct examination of cerebral spinal fluid, and immediate initiation of accurate treatment with amphotericin B and fluconazole survived patient.

[Effects of azithromycin and Chlorella vulgaris treatment on certain cytokine values and NK cell activity in an acute murine toxoplasmosis model]. / Akut toksoplazmozlu fare modelinde Azitromisin ve Chlorella vulgaris'in tedavi etkinliginin, NK aktivasyonu ve bazi sitokin degerlerinin arastirilmasi.

Toxoplasmosis is a common infection with a complicated treatment process. Azithromycin (AZT) is a macrolide antibiotic that can be effectively used in patients with cerebral and ocular toxoplasmosis and has fewer side effects. Chlorella vulgaris (CV), a single-cell green algae that contains nutrients and has various biological effects. CV extract (CVE) has been shown to have protective effects against infections via immune enhancement by increasing the cytotoxicity of NK cells, IL-12 and IFN-γ levels. The aim of this study was to investigate the protective effects of AZT and CV, individually and in combination, against acute toxoplasmosis in mice, and their effects on NK cell cytotoxixity, IL-12, IFN-γ, and IL-2 levels. Six groups of mice (Balb/c) were formed. With the exception of the healthy control (HC) group, all other groups were infected with 1 ml (11 x 104 trofozoit/ml) Toxoplasma gondii RH strain trophozoites. No further action was performed for infected control (IC) group. After 24 hours from trophozoite infection, CVE was given to CV group, AZT to azithromycin group and CVE + AZT combination to CV + AZT group by oral gavage for 6 days. All of the mice from IC, CV, AZT and CV + AZT groups were sacrified on the 8th day of the infection and serum, peritoneal fluid and spleen samples were collected. Trophozoite count of the groups were determined in all groups except HC group and the average growth inhibition activity was calculated by using the growth inhibition formula. In all groups IL-12, IFN-γ, IL-2 levels were measured with ELISA method and cytotoxicity of the NK cells were measured using Cytotox 96 Non-Radioactive Cytotoxicity Assay. The number of trophozoites were significantly lower in the CV group than the IC group (p< 0.001), and also significantly lower in CV + AZT combination group than the AZT group. According to the growth inhibition calculations CV treatment showed 88.6%, AZT treatment 98.46%, AZT + CV combination treatment 99.4% antiprotozoal activity against T.gondii compared with the IC group. NK cell cytotoxicity in the CV and the combination group were significantly higher than all the other groups (p< 0.001). IL-12 and IFN-γ levels were highest in IC group and the lowest in AZT + CV group. This situation has been linked to the fact that the severity of the infection has fallen considerably. IL-2 levels were significantly higher in CV, CV + AZT groups than in the other groups (p< 0.001). In our study, even CV administration alone caused a significant decline in infection.This may be related to the increased NK cytotoxicity, IL-2, IL-12 and IFN-γ levels. CV + AZT combination seems to be an effective treatment option than AZT alone, particularly in patients who are difficult to treat with common methods or in patients with immunosuppression.

[Effects of atovaquone and astragalus combination on the treatment and IL-2, IL-12, IFN-γ levels on mouse models of acute toxoplasmosis]. / Atovakuon ve astragalus kombinasyonunun akut toksoplazmozlu fare modeli tedavisi ve IL-2, IL-12, IFN-γ düzeyleri üzerindeki etkileri.

Reactivation of Toxoplasma gondii infections and serious clinical manifestations such as encephalitis may develop in immunocompromised subjects and AIDS patients. Different protocols are used for the treatment of toxoplasmosis in high-risk patient groups, however life-long prophylactic therapy against reactivation risk in AIDS patients may lead to several undesired results. Atovaquone is an effective antiprotozoal agent against toxoplasmosis with minor side effects. On the other hand, Astragalus membranaceus root extract (AmE) has been shown to have immunomodulatory and antimicrobial activities, empowering immunity by enhancing proliferation and activation of phagocytic cells mainly macrophages, and inducing Th1 type immune response. The aim of this study was to investigate the effectiveness of atovaquone alone and in combination with AmE, in the treatment of toxoplasmosis, and on the levels of IL-2, IL-12 and IFN-γ in experimentally infected mice with T.gondii. For this purpose, four experimental groups, each consisting of eight BALB/c mice, were set with the approval of Ethics Committee for the Animal Experiments. All the mice were infected with 0.5 ml of a suspension containing 2 x 104/ml trophozoites prepared from T.gondii RH strain by intraperitoneal injection. Twenty-four hours after the infection, atovaquone (100 mg/kg/day) was given to atovaquone group, AmE (0.075 mg/g) to astragalus group and atovaquone (100 mg/kg/day) plus AmE (0.075 mg/g) to Atovaquone + Astragalus (Ato + Astra) group by oral gavage. The mice in the fourth group, which was the control group, were all infected but untreated. The above administrations were carried out for seven days. On the 8th day peritoneal fluids of mice were collected under anaesthesia and trophozoite numbers per 1 ml were detected by counting on the Thoma slide. In addition, the heart bloods of mice were drawn and IL-2, IL-12, IFN-γ levels were determined in serum samples by using commercial ELISA kits (eBioscience, Austria). The mean number of trophozoites in Ato + Astra group was found significantly lower than the number of trophozoites in the other three groups (p< 0.05). The number of trophozoites in the atovaquone and astragalus groups were found significantly lower than the number of trophozoites in the control group (p< 0.05). There was a significant increase in IL-2 levels of astragalus group compared with the other three groups, in addition when IL-2 levels of Ato + Astra group were compared with ones in other three groups, a significant decrease was noticed (p< 0.05). There was a definite increase in IL-12 levels of atovaquone, astragalus and the control groups compared to those in Ato + Astra group (p< 0.05). A significant increase was found in IFN-γ levels in atovaquone and Ato + Astra groups compared with those in the control group (p< 0.05). Within the reach of our literature survey, this study was the first research in which the effectiveness of the combination of atovaquone and AmE was investigated in the treatment of acute toxoplasmosis. The results of our study suggested that there might be a synergy between atovaquone and AmE in the treatment of acute toxoplasmosis. In case these results are supported by further studies, atovaquone and AmE combination may have a potential to be used for therapy in immunocompromized patients such as AIDS patients who have a risk for toxoplasmosis.

Interaction between caspofungin or voriconazole and cefoperazone-sulbactam or piperacillin-tazobactam by in vitro and in vivo methods.

Immunosuppressive patients are at risk of fungal and bacterial infections. Therefore, these patients receive prophylactic, preemptive, empirical or target antifungal and concomitant antibiotic therapy. To this end, caspofungin (CAS) or voriconazole (VRC) antifungals and cefoperazone-sulbactam (CPZ/SAM) or piperacillin-tazobactam (PIP/TAZ) antibiotics may be used. Here, we aimed to investigate the interaction between these antifungals and antibiotics by in vitro and in vivo methods. The interaction was tested by chequerboard analysis and fractional inhibitory concentration index (FICI). It was also tested in a neutropenic mice-invasive candidiasis model and evaluated by fungal burden in kidney tissue of infected animals from the first day to the fifth day of treatment with 24 h intervals. A synergism was detected between CAS and CPZ/SAM (FICI = 0.1) and PIP/TAZ (FICI = 0.3). Fungal burden in tissues of drug-treated mice was reduced compared with controls in a time-dependent manner. In comparison with CAS-alone treated group, there were 1.32 log10 reductions of fungal burden in CAS + CPZ/SAM (p = 0.002) and in CAS + PIP/TAZ group (p = 0.14). The same interactions were not found with VRC and antibiotics. CPZ/SAM had stronger synergistic interaction with CAS than PIP/TAZ. The mechanism of synergism is not well understood. This is most likely due to an increase in the anticandidal effect of CAS plus antibiotics.

[Investigation of combined effectiveness of spiramycin and beta-glucan in mice models of acute toxoplasmosis and determination of IL-10, IL-12 and TNF-α levels]. / Spiramisin ve Beta-Glukan Kombinasyonu Etkinliginin Akut Toksoplazmozlu Fare Modelinde Arastirilmasi ve IL-10, IL-12 ve TNF-α Düzeylerinin Belirlenmesi.

Toxoplasmosis which is caused by Toxoplasma gondii, has a high risk of fetal infection development if the infection occurs during pregnancy. Treatment with oral spiramycin is recommended during pregnancy in order to prevent the transmission of protozoa to fetus and development of infection. Since beta- glucan is known to stimulate the immune system and increase the phagocytic activity of the cells, it has been shown to exhibit immunomodulatory effect on many infectious diseases. The objectives of this study were to investigate the effectiveness of beta-glucan alone and in combination with spiramycin and to determinate the levels of interlökin (IL)-10, IL-12 and tumor nekrosis factor (TNF)-α in mice experimentally infected with T.gondii. For this purpose, four experimental groups each consisting of eight BALB/c mice, were formed with the approval of Ethics Committee for the Animal Experiments. All the mice were infected with 2 ml of suspension containing 2 x 102/ml of trophozoite prepared from T.gondii RH strain (Refik Saydam National Public Health Agency, Parasitology Laboratory of Communicable Diseases Research Department, Ankara, Turkey), by intraperitoneal injection. Twenty-four hours after the infection, beta-glucan (3 mg/day) was given to the beta-glucan group, spiramycin (200 mg/kg/day) to the spiramycin group, beta-glucan (3 mg/day) plus spiramycin (200 mg/kg/day) to the beta-glucan-spiramycin (BG-S) group by oral gavage. The fourth group which was the control group was infected but untreated. The above administration was carried out for seven days. On the 8th day, under anaesthesia, 1 ml normal saline was given into the peritoneum, drawn back later and the number of trophozoites in 1 ml of peritoneal fluid was determined by counting them on the Thoma slide. Moreover, by drawing the heart blood; IL-10, IL-12, TNF-α levels were determined in serum samples by ELISA method (eBioscience Platinum, Austria). The number of trophozoites in the BG-S group was found significantly lower than the number of trophozoites in control, beta-glucan and spiramycin groups (p< 0.05). There was no significant difference between the beta-glucan and spiramycin groups, however the number of trophozoites in both groups was significantly lower than the number of trophozoites in the control group (p< 0.05). There was a certain decrease in IL-10 level in spiramycin and BG-S groups, compared to the control group, in addition when IL-10 levels in spiramycin and BG-S groups were compared with BG group, a significant decrease was noticed (p< 0.05). There was no difference in IL-12 levels between the groups, while there was a certain decrease in TNF-α level in beta-glucan, spiramycin, BG-S group in comparison to the control group. Within the reach of our literature survey, this study is the first research in which the effectiveness of the combination of beta-glucan and spiramycin in the treatment of acute toxoplasmosis was investigated. The results of our study suggested that there might be synergy between beta-glucan and spiramycin in the treatment of acute toxoplasmosis.

Human ocular filariasis: further evidence on the zoonotic role of Onchocerca lupi.

BACKGROUND: Among ocular vector-borne pathogens, Onchocerca volvulus, the agent of the so-called "river blindness", affects about 37 million people globally. Other Onchocerca spp. have been sporadically reported as zoonotic agents. Cases of canine onchocerciasis caused by Onchocerca lupi are on the rise in the United States and Europe. Its zoonotic role has been suspected but only recently ascertained in a single case from Turkey. The present study provides further evidence on the occurrence of O. lupi infesting human eyes in two patients from Turkey (case 1) and Tunisia (case 2). The importance of obtaining a correct sample collection and preparation of nematodes infesting human eyes is highlighted. METHODS: In both cases the parasites were identified with morpho-anatomical characters at the gross examination, histological analysis and anatomical description and also molecularly in case 1. RESULTS: The nematode from the first case was obviously O. lupi based on their morphology at the gross examination, histological analysis and anatomical description. In the second case, although the diagnostic cuticular characters were not completely developed, other features were congruent with the identification of O. lupi. Furthermore, the morphological identification was also molecularly confirmed in the Turkish case. CONCLUSIONS: The results of this study suggest that O. lupi infestation is not an occasional finding but it should be considered in the differential diagnosis of other zoonotic helminths causing eye infestation in humans (e.g., D. immitis and Dirofilaria repens). Both cases came from areas where no cases of canine onchocerciasis were previously reported in the literature, suggesting that an in depth appraisal of the infestation in canine populations is necessary. Physicians and ophthalmologists are advised on how to preserve nematode samples recovered surgically, to allow a definitive, correct etiological diagnosis.

Investigation of intestinal parasites in pig feces that are also human pathogens.

A total of 238 pig fecal specimens were collected from pig farms in Corlu (Tekirdag), Ayazma, and Arnavutköy (Istanbul) during the summer. Out of the 238 pig specimens, 105 were from pigs younger than 6 months and 133 from pigs older than 6 months. These were investigated for intestine parasites in particular the ones that are human pathogens. Cryptosporidium spp. was detected In 21 fecal specimens (8.8%), Giardia spp. in 9 (3.7%), Balantidium coli cysts in 4 (1.6%) and Ascaris suum eggs in 9 (4.1%). Giardia lamblia were found in 8 (7.6%) of 105 pigs younger than 6 months, Cryptosporidium spp. in 12 (11.4%), Balantidium coli cysts in 2 (1.5%). In the pigs older than 6 months Giardia lamblia were found in 1 (0.7%), Cryptosporidium spp. in 9 (6.7%), Balantidium coli cysts in 2 (1.5%). and Ascaris suum eggs in 9 (6.7%). The difference in the rate of G. lamblia (p=0.01) in pigs less than 6 months and of A. suum in those over 6 months was found to be statistically significant (p=0.005). Our results revealed that pigs are important sources of these parasites.

The influence of glucose added urine on the in vitro antimicrobial activity of various antibiotics.

BACKGROUND & OBJECTIVE: Factors associated with the medium, including calcium and magnesium ion concentration and pH have been shown to affect the results of susceptibility testing but very little is known about glycosuria and the effect of glucose on the antimicrobial effect of antibiotics. In this study we assessed the influence of glucose added urine on the in vitro activities of various antibiotics by the microbroth dilution method. METHODS: Sixteen Escherichia coli isolates from patients with urinary infections were used in this study. Nine antibiotics were tested for their antimicrobial activity. Minimum inhibitory concentrations (MICs) were performed by the microbroth dilution method parallel in Mueller Hinton broth and glucose added urine. RESULTS: MICs of nearly all antibiotics were higher in glucose added urine than MICs in broth. MIC(90) against ampicillin was 32-fold higher in glucose added urine than MIC(90) in broth. MIC(90)s against ampicillin-sulbactam, cephalothin, cefuroxime, ceftriaxone and ciprofloxacin in glucose added urine were significantly (P<0.05) higher than MIC(90) in broth. Equal MIC(90) in glucose added urine and broth were obtained for amikacin, sulphamethoxazole and trimethoprime. INTERPRETATION & CONCLUSION: Our findings demonstrated that MICs of antibiotics are influenced by the glucose added urine.

[Investigation of intestinal parasites in AIDS patients]. / AIDS hastalarinda barsak parazitlerinin arastirilmasi.

In this study, enteric parasites were investigated in the stool samples of 38 AIDS patients (23 with chronic diarrhea and 15 without diarrhea) prospectively. At least three stool samples from each patient were investigated microscopically for ova or trophozoites. The samples were concentrated with formol-ether method and wet preparations stained with lugol were examined. In addition, the concentrated samples were stained with modified asid-fast (Kinyoun's), rhodamine-auramine, modified trichrom and calcoflor methods. Enteric parasites were detected in 18 (47%) of the 38 patients, 16 patients harbored a single parasite, and 2 patients were found to be infected with more than one parasite. Only one (7%) of 15 AIDS patients without diarrhea, were found to be infected with Giardia lamblia. On the other hand, 17 (74%) of 23 AIDS patients with chronic diarrhea were found to be infected with various enteric parasites. Cryptosporidium spp. was detected in 9 (39%) of these 23 patients, and in 2 of them Microsporidium spp. accompanied Cryptosporidium. In 2 (9%) of these 23 patients G. lamblia were detected, while Isospora belli, Blastocystis hominis, Entamoeba histolytica, Strongyloides stercoralis and Trichuris trichiura were detected in one patient each. As a result, the detection rate of emerging parasites, including Cryptosporidium spp, Microsporidium spp, I. belli, B. hominis, and S. stercoralis was significantly higher than conventional parasites (39% versus 13%; z=2.34, p=0.01), and CD4 T cell counts were found to be significantly lower among AIDS patients with chronic diarrhea than those without diarrhea (x2=34.33, p<0.001).

The influence of urine on the in vitro antimicrobial activity of various antibiotics against different Escherichia coli phenotypes.

BACKGROUND: The influence of urine on the in vitro activities of various antibiotics used in the therapy of urinary tract infections was assessed by the microbroth dilution method in this study. METHODS: Thirty Escherichia coli strains were used: 10 E. coli strains susceptible to ampicillin, 10 strains resistant to ampicillin and ampicillin+sulbactam and ten extended spectrum beta-lactamase producer strains. The minimum inhibitory concentrations (MICs) of ampicillin, ampicillin + sulbactam, cephalothin, cefuroxime, ceftriaxone, amikacin, ciprofloxacin, sulfamethoxazole and trimethoprim were performed parallel in Mueller-Hinton broth and human urine by the microbroth dilution method. RESULTS: The MIC(90) of all antibiotics except cephalothin were higher in the urine. MICs performed in the urine were found significantly higher than those performed in broth. CONCLUSIONS: This study demonstrated that MICs of antibiotics are influenced by the human urine and that MICs of some antibiotics used in the treatment of urinary tract infections may be overestimated by the standard antibiotic testing methods.

The effects of streptozotocin-induced diabetes on brucellosis of rats.

It is believed that an infection is more common and runs a more protracted course in people with diabetes. In clinical practice, it is important to be aware of these associations, as the prognosis is often dependent upon prompt recognition and appropriate treatment. To show the course of brucellosis in the diabetic state, a model of Brucella melitensis infection was used in the setting of streptozotocin-induced diabetes in rat. B. melitensis infection proceeded more severely in diabetic rats and the severity of diabetes affected the prognosis. However, no association was found between B. melitensis and insulin using in vitro and in vivo experiments. Our study illustrates that B. melitensis infection in diabetes should be taken seriously.