The correlation between phenotypes and genotypes of macrolides, lincosamides and streptogramins B resistance in Staphylococcus aureus

For 31 clinical strains of S. aureus the correlation between phenotype and genotype of resistance to macrolides, lincosamides and streptogramins B (MLSB) was established.. Phenotypes were determined on the basis of: susceptibility to erythromycin and clindamycin and the ability to an induction of the resistance (phenotypes S, susceptible; R , constitutive resistant, D, resistant after induction with erythromycin, D+, resistant after induction with erythromycin and with a presence of the small colonies inside inhibition zone between erythromycin and clindamycin discs), and on the basis of the resistance to spectinomycin (spR, resistant, spS, susceptible). Among examined S. aureus strains eight phenotypes of resistance to MLSB were recognized (the corresponding genotypes are given in brackets). Six phenotypes were typical: SspS (lack of MLS-B resistance genes), NEGspS (msrA/B, 1 strain), D+spS (ermCi, 4 strains),. DspR (ermAi, 11 strains and ermAi + msrA/B, 2 strains), RspR (ermAc, 4 strains and ermA + msrA/B,1 strain and ermA + ermC, 1 strain) and RspS (ermCc, 6 strains and ermB, 1 strain). Two rare phenotypes in two single strains were observed: SspR (ermAi, the strain with altered inducibility, inductor other than erythromycin) and DspS (ermAi, presumably mutation or lack of spc in Tn554).     

Reduced susceptibility to vancomycin and biofilm formation in methicillin-resistant Staphylococcus epidermidis isolated from blood cultures

This study aimed to correlate the presence of ica genes, biofilm formation and antimicrobial resistance in 107 strains of Staphylococcus epidermidis isolated from blood cultures. The isolates were analysed to determine their methicillin resistance, staphylococcal cassette chromosome mec (SCCmec) type, ica genes and biofilm formation and the vancomycin minimum inhibitory concentration (MIC) was measured for isolates and subpopulations growing on vancomycin screen agar. The mecA gene was detected in 81.3% of the S. epidermidis isolated and 48.2% carried SCCmec type III. The complete icaADBC operon was observed in 38.3% of the isolates; of these, 58.5% produced a biofilm. Furthermore, 47.7% of the isolates grew on vancomycin screen agar, with an increase in the MIC in 75.9% of the isolates. Determination of the MIC of subpopulations revealed that 64.7% had an MIC ≥ 4 μg mL^-1, including 15.7% with an MIC of 8 μg mL^-1 and 2% with an MIC of 16 μg mL^-1. The presence of the icaADBC operon, biofilm production and reduced susceptibility to vancomycin were associated with methicillin resistance. This study reveals a high level of methicillin resistance, biofilm formation and reduced susceptibility to vancomycin in subpopulations of S. epidermidis. These findings may explain the selection of multidrug-resistant isolates in hospital settings and the consequent failure of antimicrobial treatment.     

Macrolide resistance genotypes and phenotypes among erythromycin-resistant clinical isolates of Staphylococcus aureus and coagulase-negative staphylococci, Italy

One hundred macrolide-resistant staphylococcal isolates from clinically relevant infections in Italy during a 19-month period were studied. Four distinct resistance phenotypes were observed using the triple-disk induction test (erythromycin, clindamycin, telithromycin): the cMLS_B phenotype (24 isolates); the iMLS_B phenotype (41 isolates); the MS phenotype (three isolates); and the iMTS phenotype (erythromycin-induced telithromycin resistance) (32 isolates). ermC and ermA genes predominated within erythromycin-resistant Staphylococcus aureus isolates with iMLS_B phenotype and cMLS_B phenotype, respectively. Among erythromycin-resistant CoNS isolates, half of the strains showed the iMTS or MS/ msrA association, and ermC gene predominated among isolates with MLS_B phenotype. By pulsed-field gel electrophoresis, high genetic heterogeneity was observed among the isolates studied. Both independent acquisition of macrolide resistance genes and spread of specific resistant clones were observed. Association between certain clonal types and specific types of infection could be detected. To our knowledge, this is the first report on characterization of erythromycin-resistant staphylococci in Italy.     

The antimicrobial susceptibility,biofilm formation and genotypic profiles of Staphylococcus haemolyticus from bloodstream infections

We analysed the antimicrobial susceptibility, biofilm formation and genotypic profiles of 27 isolates of Staphylococcus haemolyticus obtained from the blood of 19 patients admitted to a hospital in Rio de Janeiro, Brazil. Our analysis revealed a clinical significance of 36.8% and a multi-resistance rate of 92.6% among these isolates. All but one isolate carried the mecA gene. The staphylococcal cassette chromosome mec type I was the most prevalent mec element detected (67%). Nevertheless, the isolates showed clonal diversity based on pulsed-field gel electrophoresis analysis. The ability to form biofilms was detected in 66% of the isolates studied. Surprisingly, no icaAD genes were found among the biofilm-producing isolates.     

Shigella in Brazilian children with acute diarrhoea: prevalence,antimicrobial resistance and virulence genes

Diarrhoeal disease is still considered a major cause of morbidity and mortality among children. Among diarrhoeagenic agents, Shigella should be highlighted due to its prevalence and the severity of the associated disease. Here, we assessed Shigella prevalence, drug susceptibility and virulence factors. Faeces from 157 children with diarrhoea who sought treatment at the Children''s Hospital João Paulo II, a reference children´s hospital in Belo Horizonte, state of Minas Gerais, Brazil, were cultured and drug susceptibility of the Shigella isolates was determined by the disk diffusion technique. Shigella virulence markers were identified by polymerase chain reaction. The bacterium was recovered from 10.8% of the children (88.2% Shigella sonnei). The ipaH, iuc, sen and ial genes were detected in strains isolated from all shigellosis patients; set1A was only detected in Shigella flexneri. Additionally, patients were infected by Shigella strains of different ial, sat, sen and set1A genotypes. Compared to previous studies, we observed a marked shift in the distribution of species from S. flexneri to S. sonnei and high rates of trimethoprim/sulfamethoxazole resistance.     

Mutational and acquired carbapenem resistance mechanisms in multidrug resistant Pseudomonas aeruginosa clinical isolates from Recife, Brazil

An investigation was carried out into the genetic mechanisms responsible for multidrug resistance in nine carbapenem-resistant Pseudomonas aeruginosaisolates from different hospitals in Recife, Brazil. Susceptibility to antimicrobial agents was determined by broth microdilution. Polymerase chain reaction (PCR) was employed to detect the presence of genes encoding β-lactamases, aminoglycoside-modifying enzymes (AMEs), 16S rRNA methylases, integron-related genes and OprD. Expression of genes coding for efflux pumps and AmpC cephalosporinase were assessed by quantitative PCR. The outer membrane proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The blaSPM-1, blaKPC-2 and blaGES-1 genes were detected in P. aeruginosaisolates in addition to different AME genes. The loss of OprD in nine isolates was mainly due to frameshift mutations, premature stop codons and point mutations. An association of loss of OprD with the overexpression of MexAB-OprM and MexXY-OprM was observed in most isolates. Hyper-production of AmpC was also observed in three isolates. Clonal relationship of the isolates was determined by repetitive element palindromic-PCR and multilocus sequence typing. Our results show that the loss of OprD along with overexpression of efflux pumps and β-lactamase production were responsible for the multidrug resistance in the isolates analysed.     

The genus Gymnetron from China with description of pre-imaginal stages of G. miyoshii,G. auliense and G. vittipenne (Coleoptera,Curculionidae)

There are four species of Gymnetron in China recorded to date including Gymnetron miyoshii Miyoshi, 1922, Gymnetron villosipenne Roelofs, 1875, Gymnetron auliense Reitter, 1907 and Gymnetron vittipenne Marseul, 1876, of which the last two are new country records. The pre-imaginal stages including eggs, mature larvae and pupae of Gymnetron miyoshii, Gymnetron auliense and Gymnetron vittipenne are described and illustrated. In addition, their diagnostic characters (larvae and pupae) are discussed and differentiated, and notes on some of their biological parameters are provided. Potential ecological impacts between Gymnetron weevils and their host Veronica spp. also are provided.     

Resistance of Trisomic and Diploid Hybrids of Beta vulgaris and B. procumbens to the Sugarbeet Nematode,Heterodera schachtiis Arnold

Trisomic and diploid hybrids of sugarbeet (Beta vulgaris L.) X wild beet (B. procumbens Chr. Sin.) inherited the gene for resistance to Heterodera schachtii Schm. from B. procumbens. The hybrids showed partial resistance to H. schachtii, manifested in failure of larvae to reach maturity. Although significantly greater numbers of female nematodes developed on plants inoculated with populations from the Netherlands or Italy than on plants inoculated with a population from the Salinas Valley, California. the totals for all populations on resistant plants were small. Greater numbers of males than females developed on root-slice cultures of resistant hyhrids when compared to a susceptible cultivar.     

Transmission of the Pinewood Nematode, Bursaphelenchus xylophilus,to Slash Pine Trees and Log Bolts by a Cerambycid Beetle,Monochamus titillator, in Florida

Field-collected adults of the southern pine sawyer, Monochamus titillator (F.) (Coleoptera: Cerambycidae), naturally infested with fourth-stage juveniles (dauerlarvae) of the pinewood nematode, Bursaphelenchus xylophilus (Steiner and Buhrer, 1934) Nickle, 1970, were maturation fed on excised shoots of typical slash pine, Pinus elliottii Engelm. var elliottii, for 21 days. During August 1981, a male and female adult beetle were held in a sleeve cage placed on the terminal of a side branch of each of seven replicate, healthy 10-year-old slash pine trees. All seven branch terminals showed evidence of beetle feeding on the bark after 1 week, and pinewood nematodes were present in wood samples taken near these feeding sites. Four of the seven trees showed wilt symptoms in 4-6 weeks and died about 9 weeks after beetle feeding. Pinewood nematodes were recovered from the roots and trunks of the dead trees. Each of seven replicate slash pine log bolts was enclosed in a jar with a pair of the same beetles used in the sleeve cages. After 1 week, wood underlying beetle oviposition sites in the bark of all replicate log bolts was infested with the pinewood nematode.     

Positional cloning of rp2 QTL associates the P450 genes CYP6Z1, CYP6Z3 and CYP6M7 with pyrethroid resistance in the malaria vector Anopheles funestus

Pyrethroid resistance in Anopheles funestus is threatening malaria control inAfrica. Elucidation of underlying resistance mechanisms is crucial to improve the successof future control programs. A positional cloning approach was used to identify genesconferring resistance in the uncharacterised rp2 quantitative trait locus (QTL)previously detected in this vector using F6 advanced intercross lines (AIL). A113 kb BAC clone spanning rp2 was identified and sequenced revealing acluster of 15 P450 genes and one salivary protein gene (SG7-2). Contrary toA. gambiae, AfCYP6M1 is triplicated in A. funestus, whileAgCYP6Z2 orthologue is absent. Five hundred and sixty-five new singlenucleotide polymorphisms (SNPs) were identified for genetic mapping from rp2P450s and other genes revealing high genetic polymorphisms with one SNP every36 bp. A significant genotype/phenotype association was detected forrp2 P450s but not for a cluster of cuticular protein genes previouslyassociated with resistance in A. gambiae. QTL mapping using F6 AIL confirms therp2 QTL with an increase logarithm of odds score of 5. Multiplex geneexpression profiling of 15 P450s and other genes around rp2 followed byindividual validation using qRT–PCR indicated a significant overexpression in theresistant FUMOZ-R strain of the P450s AfCYP6Z1, AfCYP6Z3,AfCYP6M7 and the glutathione-s-transferase GSTe2 with respective foldchange of 11.2, 6.3, 5.5 and 2.8. Polymorphisms analysis of AfCYP6Z1 andAfCYP6Z3 identified amino acid changes potentially associated with resistancefurther indicating that these genes are controlling the pyrethroid resistance explained bythe rp2 QTL. The characterisation of this rp2 QTL significantly improvesour understanding of resistance mechanisms in A. funestus.     

A new species of Bembidion ( Ecuadion) from Ecuador (Coleoptera,Carabidae, Bembidiini), with a key to members of the georgeballi species group

A new species of ground beetle, Bembidion ricei, is described from the Andes mountains of Ecuador east of Quito. It belongs to the georgeballi species group of subgenus Ecuadion, and is most similar to Bembidion georgeballi. A key to the species of the group is provided.     

A revision of the Megachile subgenus Litomegachile Mitchell with an illustrated key and description of a new species (Hymenoptera,Megachilidae, Megachilini)

The species of Megachile subgenus Litomegachile are revised with a review of the species morphology, biology, and plant associations. A new species, Megachile pankus, is described and illustrated. Megachile mendica snowi Mitchell is elevated to species. Megachile var. nupta Cresson and Megachile texana var. cleomis Cockerell are synonymized with Megachile brevis and Megachile texana, respectively. An illustrated key for Litomegachile is also provided.     

A new species of Newportia Gervais, 1847 from Puerto Rico,with a revised key to the species of the genus (Chilopoda,Scolopendromorpha, Scolopocryptopidae)

A new species of the centipede genus Newportia, Newportia stoevisp. n., is described from Rio Encantado Cave, Puerto Rico. It differs from all congeners by having sternites distinctly margined laterally and ultimate legs bearing 4 spinous processes on both prefemur and femur, and 2 on tibia. The value of some terms used in the taxonomy of the genus have been analyzed and an amended identification key to the species of Newportia is provided.     

Polycystic echinococcosis in the state of Acre,Brazil: contribution to patient diagnosis,treatment and prognosis

The lack of knowledge regarding polycystic hydatid disease results in delayed or even incorrect diagnosis. The lack of systematic information regarding treatment also makes it difficult to assess the results and prognosis in patients with peritoneal and hepatic lesions caused by Echinococcus vogeli. Here we describe the clinical features of patients, propose a radiological classification protocol and describe a therapeutic option for the treatment of hydatid disease that previously had only been used for cases of cystic echinococcosis (Echinococcus granulosus). A prospective cohort study was initiated in 1999 and by 2009 the study included 60 patients. These patients were classified according to the PNM classification (parasite lesion, neighbouring organ invasion and metastases) and placed in one of three therapeutic modalities: (i) chemotherapy with albendazole at a dose of 10 mg/kg/day, (ii) surgical removal of cysts or (iii) percutaneous puncture of the cysts via puncture, aspiration, injection and re-aspiration (PAIR). The results were stratified according to therapeutic outcome: "cure", "clinical improvement", "no improvement", "death" or "no information". The PNM classification was useful in indicating the appropriate therapy in cases of polycystic hydatid disease. In conclusion, surgical therapy produced the best clinical results of all the therapies studied based on "cure" and "clinical improvement" outcomes. The use of PAIR for treatment requires additional study.     

Description of Trilineellus clathrocutis n.g., n.sp. (Tylenchorhynchinae: Tylenchida Thorne, 1949) with a Key to Species and Observations on Tylenchorhynchus sensu stricto

TrilineeIlus clathrocutis n.g., n.sp. is described and illustrated. It was found as an associate of corn (Zea mays) in Stockton, Georgia, USA, and is related to a group of Tylenchorhynchus sensu lato species having three lines in nonareolated lateral fields. This new species is closely related to Tylenehorhynehus divittatus Siddiqi 1961, T. sculptus Seinhorst 1963, and T. triglyphus Seinhorst 1963 (syn. T. chonai Sethi & Swarup 1968) Tarjan 1973. It differs from these species primarily by having longitudinal striae on the body. These four species are differentiated from Tylenchorhynchus sensu stricto by having three lateral lines instead of four. They differ from Uliginotylenchus Siddiqi 1971 by having nonareolated lateral fields, fewer than 25 annules on conoid rounded tails, differently shaped gubernacula, nonattenuated stylets, and other distinctive characters. They differ from Triversus Sher 1973 by having the male tail enclosed by the bursa and by having rounded female tails. SEM observations of T. clathrocutis reveal a cuticle deeply cut by longitudinal and horizontal striae and bearing wide (> 2.0 μm) annules. Trilineellus is proposed to accommodate the new species and the three-incisured species still within Tylenchorhynchus. Tylenchorhynchus is thereby the repository for species within Tylenchorhynchinae having four lines in the lateral field, no conspicuous labial disc, and bursa enclosing the male tail.     

The draft genome sequence of multidrug-resistant Pseudomonas aeruginosa strain CCBH4851, a nosocomial isolate belonging to clone SP (ST277) that is prevalent in Brazil

The high occurrence of nosocomial multidrug-resistant (MDR) microorganisms isconsidered a global health problem. Here, we report the draft genome sequence of aMDR Pseudomonas aeruginosa strain isolated in Brazil that belongsto the endemic clone ST277. The genome encodes important resistance determinantgenes and consists of 6.7 Mb with a G+C content of 66.86% and 6,347 predictedcoding regions including 60 RNAs.     

New and Known Species of Pratylenchus Filipjev, 1936 (Nematoda: Pratylenchidae) from Haryana,India, with Remarks on Intraspecific Variations

Two new monosexual and one bisexual species Pratylenchus Filipjev, 1936 collected from Haryana state of India are described and illustrated. The primary distinguishing features of these species are Pratylenchus microstylus n. sp.: L = 331-458 μm, spear = 11 or 12 μm; Pratylenchus cruciferus n. sp.: L = 648-793 μm, central core of lateral fields with oblique lines, hemizonid 2-8 annules anterior to excretory pore; Pratylenchus ekrami n. sp.: spear = 11-13 μm, spermatheca oblong, post vulval uterine sac with differentiated cells, tail with 26-40 annules, males abundant. Studies on intraspecific variations of P. cruciferus, P. ekrami, and P. coffeae (Zimmermann, 1898) Goodey, 1951 revealed that spear length and value of ''V'' are the least variable characters. Body length and size of post vulval uterine sac varies to varying degrees in different species. Shape of median bulb in P. ekrami, number of incisures in P. coffeae, and tail shape in P. ekrami and P. coffeae exhibit the greatest amount of intraspecific variations. P. zeae Graham, 1936 and P. thornei Sher & Allen, 1953 are the other species collected during the present studies.     

Ancyronyx Erichson, 1847 (Coleoptera,Elmidae) from Mindoro,Philippines, with description of the larvae and two new species using DNA sequences for the assignment of the developmental stages

Ancyronyx buhid sp. n. and Ancyronyx tamaraw sp. n. are described based on adults and larvae, matched using their cox1 or cob DNA sequence data. Additional records of Ancyronyx schillhammeri Jäch, 1994 and Ancyronyx minerva Freitag & Jäch, 2007 from Mindoro are listed. The previously unknown larva of Ancyronyx schillhammeri is also described here, aided by cox1 data. The new species and larval stages are described in detail and illustrated by SEM and stacked microscopic images. Keys to the adult and larval Ancyronyx species of Mindoro and an updated checklist of Philippine Ancyronyx species are provided. The usefulness as bioindicators, the phylogenetic relationships and biogeographic aspects affecting the distribution patterns are briefly discussed.