Survey of Wild Mammal Hosts of Cutaneous Leishmaniasis Parasites in Panamá and Costa Rica

The eco-epidemiology of American cutaneous leishmaniasis (ACL) is driven by animal reservoir species that are a source of infection for sand flies that serve as vectors infecting humans with Leishmania spp parasites. The emergence and re-emergence of this disease across Latin America calls for further studies to identify reservoir species associated with enzootic transmission. Here, we present results from a survey of 52 individuals from 13 wild mammal species at endemic sites in Costa Rica and Panama where ACL mammal hosts have not been previously studied. For Leishmania spp. diagnostics we employed a novel PCR technique using blood samples collected on filter paper. We only found Leishmania spp parasites in one host, the two-toed sloth, Choloepus hoffmanni. Our findings add further support to the role of two-toed sloths as an important ACL reservoir in Central America.     

High prevalence of Brachyspira spp. in layers kept in alternative husbandry systems associated with frequent species variations from end of rearing to slaughter

A longitudinal survey was conducted to investigate the presence of Brachyspira species in layer flocks. A total of 66 layer flocks kept in alternative husbandry systems were sampled at three time points: end of rearing, at peak of lay and at end of lay. Content from caecal samples of freshly killed birds was cultured at each sampling time point and processed for further investigations. Gross pathological lesions in caeca were recorded during post mortem investigation. Spirochaetes were isolated from 50 flocks: three flocks were positive at all three sampling points, 25 flocks at two and 22 flocks at one sampling point, respectively. The presence of Brachyspira spp. could not be related to specific gross pathological caecal lesions or antibiotic treatments. The number of positive flocks increased with the age of birds. Furthermore, organic flocks were more often positive than flocks from barn systems. In total 80 spirochaetal cultures were obtained. B. intermedia (43.8%) was the most common species, followed by B. pulli (13.8%) and B. pilosicoli (12.5%). Brachyspira murdochii and B. innocens were found in 5.0% and 2.5%, respectively, whereas 11.3% of Brachyspira isolates could not be identified to species level. In 11.3% of the samples mixed infections were detected. Finally, the longitudinal survey revealed for the first time a possible shift in the Brachyspira species in a substantial number (32%) of layer flocks during their lifetime.     

Management of severe pain after dermal contact with caterpillars (erucism): a prospective case series

Context: Erucism, envenomation caused by dermal contact with larval forms of moths, may result in intense local pain, mainly after contact with puss caterpillars (family Megalopygidae).

Objective: To evaluate the response to different treatments for controlling severe pain in a case series of erucism in Campinas, southeastern Brazil.

Patients and methods: Prospective cohort study. A Numeric Pain Rating Scale (NPRS 0–10) was used to assess pain intensity in the Emergency Department (ED). Pain was considered as severe upon ED admission (T0) when the NPRS was ≥8. Inclusion criteria: age ≥8 years old, severe pain at T0, with continuous assessment of pain intensity in all patients using the NPRS during the ED stay (T5, T15, T30, T60?min and at discharge).

Results: Fifty-five patients fulfilled the inclusion criteria and were divided into three groups according to the initial treatment at T0: local anesthesia alone with 2% lidocaine (group 1, n?=?15), local anesthesia and analgesics (group 2, n?=?26) and analgesics without local anesthesia (group 3, n?=?14). Most patients were admitted within 2?h after dermal contact with the stinging bristles of caterpillars (median =90?min, IQR: 40–125?min). In 22 cases (40%), the caterpillar was brought for identification (Podalia spp., n?=?18; Megalopyge spp., n?=?4). There was a significant decrease in pain from T5 onwards with all of the treatments. When the short-term response (T5 and T15) was considered, analgesia was more effective in groups 1 and 2 compared to group 3 (p?n?=?25/55), mainly in group 1 (n?=?11/15). The median length of stay in the ED was 120?min (IQR: 80–173?min).

Conclusions: The association of local anesthesia with analgesics was apparently a good combination for the rapid management of severe pain in the ED.     

Occurrence of extended spectrum beta-lactamase enzymes in clinical isolates of Klebsiella species from Harar region, eastern Ethiopia

Extended Spectrum beta-Lactamases (ESBLs) producer and multidrug resistant Klebsiella spp. are becoming a major nosocomial pathogen globally. There are no documented reports yet on the occurrence of ESBL enzymes in Klebsiella spp. species from Ethiopia. This study was undertaken to isolate and determine the occurrence of ESBLs and multi-drug resistant Klebsiella spp. in different clinical samples obtained from patients. A cross-sectional survey was conducted in four different hospitals of Harar region (Hiwot Fana, Misrak-Arbegnoch, Police and Army) from December 2003 to February 2004. Three hundred and eighty four clinical specimens (202 sputum, 164 urine and 18 pus) were collected from patients admitted in different wards. Antimicrobial susceptibilities were performed on 57 clinical isolates by standard disk diffusion procedures against eight antimicrobial agents. The ESBLs detection was made by using cefotaxime and ceftazidime alone and in combination with clavulanate. A total of 57 (15%) Klebsiella spp. were isolated from 384 patients. Of the 57 isolates, 33 (58%) were from sputum, 18 (31.5%) from urine and 6 (10.5%) from pus. Of the 57 Klebsiella spp., 54 (94.7%) were identified as K. pneumoniae and 3 (5.3%) as K. oxytoca. Resistance was found against cephalosporins [cefotaxime (39.0%), cefoxitin (39.0%), ceftazidime (40.0%), ceftriaxone (40.0%), cephalothin (42.0%)], chloramphenicol (70.0%), gentamicin (61.0%) and trimethoprim-sulphamethoxazole (65.0%). Analyzed Klebsiella isolates were characterized also by a high degree of multi-resistance (67.0%). In 19/57 (33.3%) of the Klebsiella isolates, ESBL production was detected. Rates of detection of ESBL producers were 42.1, 26.3, 26.3 and 5.3% in Hiwot-Fana, Misrak-Arbegnoch, Police and Army hospitals, respectively. Multi-drug resistant isolates were more prevalent among the ESBLs producers (95.0%) than non-producers (53.0%) (p=0.24). In conclusion, our results show that awareness of ESBL production by Klebsiella spp. is clinically important. In the absence of infection control measures, ESBL producing organisms readily pass horizontally from patient to patient. These strains also transiently colonize the hands of hospital staff members, thereby facilitating patient-to-patient transmission of the organism.     

Evaluation of nested polymerase chain reaction for the early detection of Leptospira spp. DNA in serum samples from patients with leptospirosis

The aim of this study was to analyse the nested polymerase chain reaction (nested PCR) in human serum samples of patients with clinical manifestations of leptospirosis. The cases of leptospirosis were defined by the microagglutination test (MAT). The samples were collected in 2010. Of 1042 serum samples collected from 521 patients, 28 (5.4%) were considered positive cases of leptospirosis, and 493 (94.6%) were negative. Twenty-three confirmed cases had no MAT-detectable antibodies in the acute sample (mean of 5.6 days after onset). Nested PCR was positive in 22/23 (95.7%) patients during the acute phase of the disease, with negative results by MAT. Nested PCR was negative in all convalescent serum samples with positive results by MAT. All negative cases of leptospirosis were negative by nested PCR. The nested PCR is an alternative diagnostic tool for early detection of leptospires in sera during the first 7 days of the disease.     

北京市集贸市场生鲜猪肉肠球菌的耐药特征分析

  目的  探讨鸡蛋蛋壳表面肠球菌的存在情况和耐药性。  方法  从养鸡场取新鲜鸡蛋,在蛋壳表面进行擦拭,叠氮钠 - 结晶紫 - 七叶苷琼脂（BEAA）进行分离培养,16S rDNA通用引物进行PCR扩增、产物测序,对分离的肠球菌进行耐药性检测。  结果  在160个样品中分离到5种共124株肠球菌,分离率为77.5 %,主要为粪肠球菌（58.1 %）和屎肠球菌（13.1 %）,其次为鸟肠球菌（5.0 %）、鸡肠球菌（1.9 %）、铅黄肠球菌（1.2 %）。耐药性检测结果表明肠球菌对14种抗生素表现出不同的耐药性,耐药谱广,最多发现9重耐药菌。  结论  新鲜鸡蛋肠球菌携带率较高,其耐药性严重,应引起相关部门重视。     

志贺菌耐多药外排泵emrE基因作用分析

目的 对临床耐多药志贺菌株H24的1种小多重耐药性家族(SMR)外排泵emrE基因进行定向分子进化,研究基因突变增强emrE外排泵耐药性的潜力,调查志贺菌外排泵基因emrE基因垂直进化对临床耐药发展的影响。方法 通过用体外易错PCR(EP-PCR)方法对emrE基因进行随机突变实验,用微量稀释生长曲线法进行菌株的耐药性筛选,用荧光实时定量RT-PCR测定基因表达情况,应用DNA分析软件DNAStar进行生物信息学分析。结果 在31个突变子中筛选到了突变子ep2-emrE(DH5α),它的emrE基因有6个氨基酸突变,分别为Thr-28→Ala,Cys-39→Ser,Tyr-40→终止子,Ser-72→Arg,Leu-93→Phe,Ile-101→Phe。它不但增强四环素和红霉素的耐药能力,还产生新的氯霉素耐药能力。结论 尽管emrE基因较易获得新增耐药性,但在志贺菌对四环素、红霉素等抗生素耐药性发展中,emrE基因的进化突变没有被筛选,表明有其他能力更强的耐药基因在突变进化中起作用。