DDT and pyrethroid resistance status and laboratory evaluation of bio-efficacy of long lasting insecticide treated nets against Culex quinquefasciatus and Culex decens in Ghana.

Nuisance from Culex mosquitoes in Ghana has a serious negative impact on the standard of living in many urban communities. In addition, a perceived lack of efficacy of long lasting insecticide treated nets (LLINs) against nuisance mosquitoes contributes to their discontinued use. This again compromises malaria control, even if Anopheles species themselves would still be susceptible to the insecticides used. Control strategies involve pyrethroid insecticides but information on Culex mosquito susceptibility to these insecticides is limited. A nationwide survey was conducted to address this problem. In adults, susceptibility to permethrin, deltamethrin and DDT as well as enzyme activity and kdr mutation were determined. Cone and tunnel bioassay were also carried out to determine the efficacy of LLINs against the mosquitoes. Culex quinquefasciatus and Culex decens were identified in the study area. Higher deltamethrin and DDT resistance and relatively low permethrin resistance were observed in both species. High enzyme activities and kdr mutations were observed in C. quinquefasciatus but not in C. decens. However, reduced efficacy of LLINs was observed in both mosquito species. This adds up to the evidence of the spread of pyrethroid resistance in mosquitoes and its negative impact on control strategies.

Preliminary assessment of the potential role of urbanization in the distribution of carbamate and organophosphate resistant populations of Culex species in Ghana.

BACKGROUND: Besides its role as a pathogen vector, Culex species also indirectly promotes the transmission of malaria if the use of bed nets or indoor residual spraying is discontinued due to a lack of insecticide efficacy against it. A recent survey revealed widespread occurrence of pyrethroid resistance among urban populations of this mosquito in Ghana. In order to plan and implement insecticide-based resistance management strategies, this study was carried out to assess resistance status of Culex species to organophosphate and carbamate in urban areas in Ghana and the possible mechanisms involved as well as environmental factors associated with its distribution. METHODS: Mosquito larvae were sampled from various land use and ecological settings and in different seasons. In adults, susceptibility to organophosphates (fenitrothion, malathion) and carbamates (propoxur, bendiocarb) were determined. Mixed function oxidase (MFO) and α- and ß-esterase assays, as well as a PCR diagnostic assay to determine ace1 mutation were performed in individual mosquitoes. RESULTS: Culex quinquefasciatus as well as C. decens and other unidentified Culex species were found breeding in polluted water bodies in the study sites. Across all sites and seasons, carbamate induced mortality was 94.1% ± 15.4 whereas mortality caused by organophosphate was 99.5% ± 2.2. In addition, ace1 mutation and high levels of esterases were detected in some of the mosquito populations. There was a strong correlation between susceptibility status of the mosquitoes and the level of absorbance of ß-esterase (Pearson r=- 0.841, p=0.004). CONCLUSIONS: The study found low prevalence of resistance to carbamate and organophosphate insecticides among Culex species from Ghana. However, there were populations with ace1 mutations and high levels of esterases, which can confer high resistance to these classes of insecticides. Thus, it is important to monitor activities or behaviour that has the potential to select for carbamate and organophosphate resistance populations.

Value of histopathology, immunohistochemistry, and real-time polymerase chain reaction in the confirmatory diagnosis of Encephalitozoon cuniculi infection in rabbits.

Morphological lesions in kidneys and brain are all too often considered diagnostic for confirmation of encephalitozoonosis in rabbits. The current study evaluated the diagnostic value of histology versus other etiological tests, including immunohistochemistry and real-time polymerase chain reaction (PCR) for Encephalitozoon cuniculi infection diagnosis. Samples of brain, heart, lungs, intestine, liver, and kidneys from 81 rabbits were examined for morphological lesions attributed to E. cuniculi infection as well as for the presence of spores and E. cuniculi antigen. Of these, 55 rabbits were tested for E. cuniculi DNA. Histological changes consistent with E. cuniculi infection were seen in 33 rabbits (41%, 33/81) representing 87% (33/38) of all rabbits with confirmed E. cuniculi infection. Brains of these rabbits displayed 6 different types of focal lesions corresponding to the stage of infection and specific tissue response. In 5 rabbits that were tested positive, histology was either inconclusive or inconspicuous. Etiological diagnosis was based on histological spore detection in 16% (6/38) of infected rabbits. Immunohistochemistry was more sensitive (42%, 16/38) than histological spore detection, and real-time PCR proved to be the most sensitive of all investigated methods (30/35, 86% of the examined rabbits with E. cuniculi infection). Encephalitozoon cuniculi infection rarely occurs without characteristic kidney and brain lesions. However, the spectrum of brain changes is wider than previously reported. Based on these findings, confirmation of pathogenic E. cuniculi infection should include standard histology of the predilection sites and a specific etiological assay, preferably real-time PCR.

Treatment of tuberculosis in a region with high drug resistance: outcomes, drug resistance amplification and re-infection.

INTRODUCTION: Emerging antituberculosis drug resistance is a serious threat for tuberculosis (TB) control, especially in Eastern European countries. METHODS: We combined drug susceptibility results and molecular strain typing data with treatment outcome reports to assess the influence of drug resistance on TB treatment outcomes in a prospective cohort of patients from Abkhazia (Georgia). Patients received individualized treatment regimens based on drug susceptibility testing (DST) results. Definitions for antituberculosis drug resistance and treatment outcomes were in line with current WHO recommendations. First and second line DST, and molecular typing were performed in a supranational laboratory for Mycobacterium tuberculosis (MTB) strains from consecutive sputum smear-positive TB patients at baseline and during treatment. RESULTS: At baseline, MTB strains were fully drug-susceptible in 189/326 (58.0%) of patients. Resistance to at least H or R (PDR-TB) and multidrug-resistance (MDR-TB) were found in 69/326 (21.2%) and 68/326 (20.9%) of strains, respectively. Three MDR-TB strains were also extensively resistant (XDR-TB). During treatment, 3/189 (1.6%) fully susceptible patients at baseline were re-infected with a MDR-TB strain and 2/58 (3.4%) PDR-TB patients became MDR-TB due to resistance amplification. 5/47 (10.6%) MDR- patients became XDR-TB during treatment. Treatment success was observed in 161/189 (85.2%), 54/69 (78.3%) and 22/68 (32.3%) of patients with fully drug susceptible, PDR- and MDR-TB, respectively. Development of ofloxacin resistance was significantly associated with a negative treatment outcome. CONCLUSION: In Abkhazia, a region with high prevalence of drug resistant TB, the use of individualized MDR-TB treatment regimens resulted in poor treatment outcomes and XDR-TB amplification. Nosocomial transmission of MDR-TB emphasizes the importance of infection control in hospitals.

High prevalence of Capillaria plica infections in red foxes (Vulpes vulpes) in Southern Germany.

The nematode Capillaria plica is an ubiquitous parasite of the urinary tract of Canidae and Felidae. It causes a wide spectrum of clinical symptoms, ranging from asymptomatic infections to urinary bladder inflammation, pollacisuria, dysuria, and hematuria. Foxes serve as reservoir hosts and are considered to be a potential source of infection for companion and hunting dogs as well as domestic cats which acquire the infection by ingestion of earthworms which are the intermediate hosts. Despite its importance, epidemiological studies on this parasite are scarce and almost entirely lacking altogether for Central Europe. Therefore, we examined 116 red foxes (Vulpes vulpes) for the infection of C. plica by pathologic examination of the urinary bladders and microscopy of mucosal smears and urine sediments. The parasite was detected in 90 (78%; 95% CI, 68.9-84.8%) of the foxes, originating from all administrative districts of Bavaria (Southern Germany). Since Bavaria is characterized by a high number of forests and wildlife sanctuaries that provide ideal living conditions for foxes, the corresponding risk of infection of companion and hunting dogs by oral ingestion of earthworms as the intermediate hosts can likewise not be excluded. Because of the scarcity of reports on prevalences of C. plica worldwide, we also include a brief review of the available literature.

Characteristics of drug-resistant tuberculosis in Abkhazia (Georgia), a high-prevalence area in Eastern Europe.

Although multidrug-resistant (MDR) tuberculosis (TB) is a major public health problem in Eastern Europe, the factors contributing to emergence, spread and containment of MDR-TB are not well defined. Here, we analysed the characteristics of drug-resistant TB in a cross-sectional study in Abkhazia (Georgia) between 2003 and 2005, where standard short-course chemotherapy is supplemented with individualized drug-resistance therapy. Drug susceptibility testing (DST) and molecular typing were carried out for Mycobacterium tuberculosis complex strains from consecutive smear-positive TB patients. Out of 366 patients, 60.4% were resistant to any first-line drugs and 21% had MDR-TB. Overall, 25% of all strains belong to the Beijing genotype, which was found to be strongly associated with the risk of MDR-TB (OR 25.9, 95% CI 10.2-66.0) and transmission (OR 2.8, 95% CI 1.6-5.0). One dominant MDR Beijing clone represents 23% of all MDR-TB cases. The level of MDR-TB did not decline during the study period, coinciding with increasing levels of MDR Beijing strains among previously treated cases. Standard chemotherapy plus individualized drug-resistance therapy, guided by conventional DST, might be not sufficient to control MDR-TB in Eastern Europe in light of the spread of "highly transmissible" MDR Beijing strains circulating in the community.

Genetic evidence for multi-event imports of avian influenza virus A (H5N1) into Bavaria, Germany.

The almost simultaneous initial detections of avian influenza A H5N1 viruses in central Europe in February 2006, at a time devoid of migratory bird activity, raised the question of the origin of these viruses. This report presents molecular data from Europe providing evidence for multiple and spatially overlapping H5N1 introductions into Bavaria, Germany.

Disseminated lethal Encephalitozoon cuniculi (genotype III) infections in cotton-top tamarins (Oedipomidas oedipus)--a case report.

For the first time, Encephalitozoon (E.) cuniculi genotype III ('dog strain') was verified in two cotton-top tamarins (Oedipomidas oedipus) by light microscopy, immunohistochemistry, electron microscopy, PCR and sequencing. The animals had a disseminated lethal infection with this protist. In earlier reports, genotype III had been found only in domestic dogs, man, emperor tamarins (Saguinus imperator) and golden lion tamarins (Leontopithecus rosalia). This investigation establishes now that the 'dog strain' can occur in cotton-top tamarins too. This is further evidence for the zoonotic potential of E. cuniculi. Furthermore, free E. cuniculi spores were identified also in blood vessels of several tissues. These findings indicate that during a disseminated infection E. cuniculi spores can occur in peripheral blood, too. We propose that blood should also be included in the investigations for the detection of microsporidia, so that a possible disseminated course of an infection can be detected.

"Staphylococcus pettenkoferi," a novel staphylococcal species isolated from clinical specimens.

In this report we describe a novel species of coagulase-negative novobiocin susceptible staphylococci obtained from an epidemiologically unrelated blood culture and a wound infection. These isolates significantly differed from all other validated Staphylococcus species based on phenotypic characteristics and 16S rRNA gene sequencing. Both isolates had identical 16S rRNA sequences and phylogenetic trees constructed from evolutionary distances showed that this species formed a distinct and deep subline that was most closely related to members of the Staphylococcus saprophyticus cluster group (S. kloosii, S. gallinarum, S. arlettae, S. saprophyticus, S. xylosus, S. equorum, S. succinus and S. cohnii) and Staphylococcus auricularis. Furthermore these strains could each be distinguished from all other staphylococci based on at least one phenotypic trait. Therefore we propose the designation of "Staphylococcus pettenkoferi" a novel species of coagulase-negative staphylococci.

First detection of the microsporidium Enterocytozoon bieneusi in non-mammalian hosts (chickens).

Faecal samples taken from eight underweight, approximately 5-week-old broiler chickens in a poultry abattoir were investigated for microsporidial infections by light microscopy, electron microscopy, and PCR. In two of six chickens, which were suspected of being infected with microsporidia by light microscopy, Enterocytozoon bieneusi (genotype 'J') was detected by PCR and DNA sequencing, and in one of the two PCR-positive samples by extensive electron microscopy. This is the first time that E. bieneusi has been detected in chickens, i.e. in a non-mammalian species.