Comparative molecular and microbiologic diagnosis of bacterial endocarditis

Sequencing of 16S rDNA, and of sodAint and rpoBint in some cases, was applied to DNA from heart valves of 46 patients (36 with definite and 10 with possible endocarditis). Sequence-based identifications were compared with those obtained with conventional methods. Among the 36 definite cases, 30 had positive blood cultures and 6 had negative cultures. Among the 30 positive cases, sequencing of 16S rDNA permitted identification of species (18), genus (8), or neither (4); sodAint and rpoBint sequencing was necessary for species identification in 8 cases. Species identifications were identical in only 61.5%, when conventional techniques and DNA sequencing were used. In five of the six blood culture-negative endocarditis cases, sequencing identified Bartonella quintana (3), B. henselae (1), and Streptococcus gallolyticus (1). Our results demonstrate a clear benefit of molecular identification, particularly in cases of blood culture-negative endocarditis and of possible endocarditis, to confirm or invalidate the diagnosis. Moreover, in 19.4% of the definite cases, the improvement in species identification by sequencing led to improved patient management.     

Emergence of Raoultella ornithinolytica on O‘ahu: A Case of Community-acquired R. ornithinolytica Urinary Tract Infection

Human infection with Raoultella ornithinolytica is rare, with only ten cases having been reported previously. This case report describes a local patient diagnosed with community-acquired R. ornithinolytica urinary tract infection in 2014.     

Tropheryma whipplei Endocarditis

Tropheryma whipplei endocarditis differs from classic Whipple disease, which primarily affects the gastrointestinal system. We diagnosed 28 cases of T. whipplei endocarditis in Marseille, France, and compared them with cases reported in the literature. Specimens were analyzed mostly by molecular and histologic techniques. Duke criteria were ineffective for diagnosis before heart valve analysis. The disease occurred in men 40–80 years of age, of whom 21 (75%) had arthralgia (75%); 9 (32%) had valvular disease and 11 (39%) had fever. Clinical manifestations were predominantly cardiologic. Treatment with doxycycline and hydroxychloroquine for at least 12 months was successful. The cases we diagnosed differed from those reported from Germany, in which arthralgias were less common and previous valve lesions more common. A strong geographic specificity for this disease is found mainly in eastern-central France, Switzerland, and Germany. T. whipplei endocarditis is an emerging clinical entity observed in middle-aged and older men with arthralgia.     

Epidemiology,Clinical Manifestations,and Outcomes of Streptococcus suis Infection in Humans

Streptococcus suis, a bacterium that affects pigs, is a neglected pathogen that causes systemic disease in humans. We conducted a systematic review and meta-analysis to summarize global estimates of the epidemiology, clinical characteristics, and outcomes of this zoonosis. We searched main literature databases for all studies through December 2012 using the search term “streptococcus suis.” The prevalence of S. suis infection is highest in Asia; the primary risk factors are occupational exposure and eating of contaminated food. The pooled proportions of case-patients with pig-related occupations and history of eating high-risk food were 38.1% and 37.3%, respectively. The main clinical syndrome was meningitis (pooled rate 68.0%), followed by sepsis, arthritis, endocarditis, and endophthalmitis. The pooled case-fatality rate was 12.8%. Sequelae included hearing loss (39.1%) and vestibular dysfunction (22.7%). Our analysis identified gaps in the literature, particularly in assessing risk factors and sequelae of this infection.     

Bats as Reservoir Hosts of Human Bacterial Pathogen,Bartonella mayotimonensis

A plethora of pathogenic viruses colonize bats. However, bat bacterial flora and its zoonotic threat remain ill defined. In a study initially conducted as a quantitative metagenomic analysis of the fecal bacterial flora of the Daubenton’s bat in Finland, we unexpectedly detected DNA of several hemotrophic and ectoparasite-transmitted bacterial genera, including Bartonella. Bartonella spp. also were either detected or isolated from the peripheral blood of Daubenton''s, northern, and whiskered bats and were detected in the ectoparasites of Daubenton''s, northern, and Brandt''s bats. The blood isolates belong to the Candidatus-status species B. mayotimonensis, a recently identified etiologic agent of endocarditis in humans, and a new Bartonella species (B. naantaliensis sp. nov.). Phylogenetic analysis of bat-colonizing Bartonella spp. throughout the world demonstrates a distinct B. mayotimonensis cluster in the Northern Hemisphere. The findings of this field study highlight bats as potent reservoirs of human bacterial pathogens.     

Methicillin-Resistant Staphylococcus aureus (MRSA) Detected at Four U.S. Wastewater Treatment Plants

Background: The incidence of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infections is increasing in the United States, and it is possible that municipal wastewater could be a reservoir of this microorganism. To date, no U.S. studies have evaluated the occurrence of MRSA in wastewater.Objective: We examined the occurrence of MRSA and methicillin-susceptible S. aureus (MSSA) at U.S. wastewater treatment plants.Methods: We collected wastewater samples from two Mid-Atlantic and two Midwest wastewater treatment plants between October 2009 and October 2010. Samples were analyzed for MRSA and MSSA using membrane filtration. Isolates were confirmed using biochemical tests and PCR (polymerase chain reaction). Antimicrobial susceptibility testing was performed by Sensititre® microbroth dilution. Staphylococcal cassette chromosome mec (SCCmec) typing, Panton-Valentine leucocidin (PVL) screening, and pulsed field gel electrophoresis (PFGE) were performed to further characterize the strains. Data were analyzed by two-sample proportion tests and analysis of variance.Results: We detected MRSA (n = 240) and MSSA (n = 119) in 22 of 44 (50%) and 24 of 44 (55%) wastewater samples, respectively. The odds of samples being MRSA-positive decreased as treatment progressed: 10 of 12 (83%) influent samples were MRSA-positive, while only one of 12 (8%) effluent samples was MRSA-positive. Ninety-three percent and 29% of unique MRSA and MSSA isolates, respectively, were multidrug resistant. SCCmec types II and IV, the pvl gene, and USA types 100, 300, and 700 (PFGE strain types commonly found in the United States) were identified among the MRSA isolates.Conclusions: Our findings raise potential public health concerns for wastewater treatment plant workers and individuals exposed to reclaimed wastewater. Because of increasing use of reclaimed wastewater, further study is needed to evaluate the risk of exposure to antibiotic-resistant bacteria in treated wastewater.     

Unique Clone of Coxiella burnetii Causing Severe Q Fever,French Guiana

Acute Q fever is an emergent and severe disease in French Guiana. We obtained 5 Coxiella burnetii isolates from samples of patients from Cayenne and found an epidemic clone circulating in Cayenne. This clone has caused pneumonia and endocarditis and seems to be more virulent than previously described strains.     

Parasitic Infection of an Endemic Fish (Blicca bjoerkna) and an Exotic Fish (Hemiculter beucisculus) In Anzali Lagoon,Caspian Sea,Iran

Background

In Anzali Lagoon, there are some endemic and exotic fishes. The present study was conducted to compare the parasitic fauna of Blicca bjeorkna, as an endemic fish and Hemiculter leucisculus, as an introduced fish to the lagoon.

Methods

A parasitological investigation was done on 78 specimens of B. bjoerkna and 114 of H. leucisculus. The fishes were collected from August 2009 to April 2010 by the electro fishing from Anzali Lagoon.

Results

Eleven parasites species were found in 192 fish samples. The prevalence and mean intensity of parasites in each host were as follows: Parasites from B. bjorkna were Trichodina perforata (53.85%); Myxobolus musayevi (27.19%, 1±0.79); Dactylogyrus difformis (88.05%, 8±7.24) and D. sphyrna (5.18%, 0.95±0.51), Diplostomum spataceum (98.72%, 9.51±9.01), Posthodiplostomum cuticula (15.38%, 4.25±2.5), Ripidocotyle sp. (1.28%, 2±0.74); Contracaecum osculatum (17.95%, 1.64±0.79), Philometra rischta (12.8%, 1.4±0.54), and Raphidascaris acus (1.04%, 0.03±0.26). The H. leucisculus were infected with T. perforata (27.19%), D. spataceum (7.89%, 1.33±0.54), Ps. tomentosa (7.02%, 1.62±0.49) and R. acus (0.88%, 3±0.28). B. bjoerkna was presented as a new host for M. musayevi and C. osculatum, while H. leucisculus was introduced as a new host for T. perforata and Ps. tomentosa.

Conclusion

The prevalence of parasites was significantly more in native fish than that of exotic fish (P<0.05). This reduction in parasitic infection in H. leucisculus may be due to its immune system resistance, well adaptation to the new environment, host-specific limitation for endemic parasites and disability of introduced parasite to complete its life cycle in the new host as well.     

TEM and SHV Genes in Klebsiella pneumoniae Isolated from Cockroaches and Their Antimicrobial Resistance Pattern

Objectives

Klebsiella pneumoniae is a gram-negative rod bacterium, a known cause of community-acquired bacterial pneumonia and is an important hospital-acquired pathogen that causes severe morbidity and mortality. The aim of this study was to identify the TEM and SHV genes in K. pneumoniae isolated from cockroaches obtained from hospitals.

Methods

In this study, 250 cockroaches were collected from different hospitals in the province of Chaharmahal Va Bakhtiari, which is located in southwest Iran. The samples were examined for the presence of K. pneumoniae by plating onto a combination of culture media, and the antimicrobial susceptibility patterns of isolated K. pneumoniae from samples were evaluated using the disk diffusion test. In addition, from the culture, genomic bacterial DNA was extracted, and sequence-specific targets (TEM and SHV genes) were amplified using the polymerase chain reaction (PCR) method.

Results

Out of 250 cockroach samples collected from various hospitals, 179 samples (71.60%) were positive for K. pneumoniae. PCR reaction was performed using specific oligonucleotide primers (TEM-F, TEM-R and SHV-F, SHV-R) for the amplification of each gene, and amplified products were visualized on 1% agarose gel electrophoresis. Of all the specimens amplified by PCR in this research, 32 samples (17.87%) were positive for TEM and 15 samples (8.37%) were positive for SHV.

Conclusion

Detection of TEM and SHV genes using molecular methods and their pattern of antimicrobial resistance can provide useful information about the epidemiology of and risk factors associated with K. pneumoniae infection.     

Molecular Epidemiology of Cryptosporidiosis in Iranian Children,Tehran, Iran

Background

Cryptosporidium is a worldwide protozoan parasite and one of the most common causes of infection and diarrhea in humans and cattle. The aim of the present study was determination of subtypes of Cryptosporidium among children with diarrhea in Tehran by sequence analysis of the highly polymorphic 60-kDa glycoprotein (GP60) gene.

Methods

Fecal samples were collected from 794 diarrheic children. Initial identification of Cryptosporidium was carried out on stool samples by Ziehl-Neelsen acid-fast staining method. DNA was extracted from positive microscopically samples and Cryptosporidium genotypes and subtypes were determined, accordingly.

Results

Out of 794 collected samples, 19 (2.40%) were positive for Cryptosporidium oocysts. Sequences analysis of GP60 gene showed that 17 (89.47%) of the positive isolates were Cryptosporidium parvum and 2 (10.52%) were C. hominis. All subtypes of C. parvum isolates belonged to allele families IIa (6/17) and IId (11/17). The most common allele in all 17 isolates belonged to IId A20G1a (41.18%). A22G1 (IF) subtype was detected in two C. hominis isolates of the children.

Conclusion

The predominancy of C. parvum species (specially, IId A20G1a subtype) in current study underlines the importance of zoonotic Cryptosporidium transmission in Iran.     

Helminth Infections in Rattus ratus and Rattus norvigicus in Tehran,Iran

Background

The aim of this study was to determine intestinal and liver helminth infections in Rattus rodents in Tehran Iran.

Methods

Overall, 306 traps were put in 39 different regions in Tehran from 2009 to 2010. Rodents, including R. rattus and R. norvegicus were caught by live-traps. They become unconscious and the spinal cords were cut, afterwards the body was dissected and the stomach, small intestine, large intestine, liver, and cecum were studied separately. The dominant type and the prevalence rate of parasites in the rodents were determined based on the infected parts of their body.

Results

After recognition of the helminthes’ types, among the 120 total number of rodents, 39 belonged to males, while among the infected rodents, 57(47.5%) were female and 18(15%) were male. The prevalence of infection in Tehran was 62.5%. Seventy cases (58.33%) of helminth infections were observed in R. rattus and 5 cases (4.16%) were observed in R. norvegicus. The maximum prevalence (15.5%) was seen in the center and east part of Tehran, while the minimum (9.16%) was in the north part of the city. The helminthes types and the corresponding percentages were Hymenolepis nana fraterna (35.8%), Heterakis spumosa (17.5%), Hymenolepis diminuta (7.5%) and Capillaria annulosa (1.6%). The dominant rodent was Rattus rattus and among the identified helminthes, Hymenolepis diminuta and Hymenolepis nana fraterna are zoonotic ones.

Conclusion

The information presented here improves our understanding of the major parasitic infections that rodents harbor and can transmit to human and animal populations in Iran. To prevent infectivity of human, the hazard of the identified zoonotic species needs to be contemplated.     

Radular Teeth Morphology in Limax (Caspilimax) keyserlingi (Martens, 1880) and Parmacella ibera (Eichwald, 1841) from Northern Iran

Background

Slugs have been known worldwide as important pests of agricultural and horticultural production. They also play a role as intermediate or definitive hosts of helminths parasite. In this purpose, current study was carried out to examine slug radular teeth structure and slug infection with helminths larvae in north of Iran.

Methods

A total number of 114 slugs were collected from center and east parts of Mazandaran province from May 2011 to June 2012. The specimens were rinsed, measured, and identified. The radula of all collected slugs was extracted and stained by using Mallory II. For detecting the helminths parasite infection, foot- head and viscera of examined slugs were removed, minced, and digested with 4.5% acid pepsin.

Results

Two species of Limax (Caspilimax) keyserlingi (Martens 1880) (11.4%, 13/114) and Parmacella ibera (Eichwald 1841) (88.6%, 101/114) were prevalent in the region. There was significant difference between body length and shell size. P. ibera had the highest number of teeth rows (145±5). The radular teeth formula was approximately similar in both identified slugs. In P. ibera, there was no significant difference in the average length and width of radula. The radular teeth in L. keyserlingi were larger and thicker than P. ibera. In all examined slugs for helminths larvae infection, P. ibera (7.69%, 1/13) was infected with Strongyloid larvae from Fereidonkenar area.

Conclusion

Two prevalent species of slugs exist in the same region of which P. ibera has capability to play a role as intermediate host of nematode helminths. Radular morphology within the slug species may be also systemically informative.     

Helminth Infections of House Mouse (Mus musulus) and Wood Mouse (Apodemus sylvaticus) from the Suburban Areas of Hamadan City,Western Iran

Background

To determine the prevalence and intensity of helminths and their zoonotic importance in small rodents inhabiting in the suburban areas of Hamadan City, Iran.

Methods

The present survey was conducted on the helminth infections of two species of rodents Apodemus sylvaticus (n=60) and Mus musculus(n=72) in the suburban areas of Hamadan City during 2010-2012. Rodents were collected and examined for helminth in the different organs. The nematodes were collected in 5% formalin solution and cleared in lactophenol, cestodes and trematodes collected from intestine fixed in AFA solution and stained by acetocarmine, cleared in xylol for identification.

Results

Helminths found in A. sylvaticus and M. musculus and their prevalence for the first time in suburban areas of Hamadan City were as follows; In A. sylvaticus: Cysticercus fasciolaris(3.33%), Syphacia fredrici(26.67%), S. stroma(8.33%), Anoplocephalidae sp. (1.67%), Skrjabinotaenia lobata(5%), Plagiorchis muris(1.67%) and in M. musculus:Hymenolepis nana (16.67%), H.diminuta (5.55%), S. obvelata(30.56%), S. ohtarom (9.72%), Rodentolepis crassa (1.39%), C. fasciolaris (1.39%). Among 11 species in two rodents 4 species including S. obvelata, H. nana, H.diminuta,and P. muris have zoonotic importance. Statistically the relation between gender and their helminth infections was not significant in either M. musculus or A. sylvaticus (P>0.05).

Conclusion

This study reports 11 species of helminths and on the other hand 3 species were identified for the first time in Iran and 5 species of them have potential health importance for public health and cat.     

Two New Species of the Genus Pallisentis Van Cleave, 1928 (Acanthocephala: Quadrigyridae) from the Intestine of Channa punctatus (Bloch, 1793) from the River Gomti at Lucknow,India

Background:

Acanthocephalans are fish parasites of worldwide distribution, penetrate their thorny proboscis into the intestinal wall of host and absorb nutrients. No diagnostic tool is available except postmortem investigations and identification by parasitologists. The aim of present study was to explore and assign taxonomical status to Pallisentis species prevalent in food fishes of river Gomti, Lucknow, India.

Methods:

A survey of fishes of river Gomti was carried out during the year 2011–2013. Acanthocephalans recovered from the intestine of Channa punctatus were kept in refrigerator for eversion of proboscis, fixed in A.F.A. fixative (50% alcohol, formalin and acetic acid in ratio of 100: 6: 2.5) for 24 hours further preserved in 70% ethanol. Camera Lucida diagrams of acetoalum carmine stained permanent mounts were made for morphometric studies.

Results:

Two new species of genus Pallisentis were identified and named as P. channai n. sp. and P. vinodai n. sp., their taxonomical status is based on major characters of proboscis hooks, spines of collar and trunk region, cement gland nuclei. On average 9 fishes were found infected with Pallisentis spp. out of 60 fishes examined randomly.

Conclusion:

Pallisentis spp. are important parasitic infection in Channidae fishes with the prevalence rate of 15%. Two new species of Pallisentis recognized from Channa punctatus of river Gomti, Lucknow, India and diagnostic features of genus are given.     

Incidence of Giardia lamblia Subspecies by PCR-RFLP in Stool Specimens of Hospitalized Children at Urmia Mutahhari Hospital,West Azerbaijan Province,Iran

Background

Giardia lamblia is one of the most prevalent intestinal flagellate protozoa that infects a wide range of vertebrate hosts causing severe intestinal disorder in children.This study was performed to determine subspecies of G.lamblia by the PCR-RFLP method, targeting the glutamate dehydrogenase(gdh)locus, in hospitalized children at Urmia Mutahhari Hospital, West Azerbaijan Province,Iran and determining the infection transformational storages in this area.

Methods

Overall, 720 stool specimens were collected from the hospitalized children, 34 samples were positive and Giardia cysts were detected under the microscope. Cysts were partially purified by the sucrose density gradient method and then washed with sterile distilled water to remove effectively the PCR inhibitors. Genomic DNA of G. lamblia isolates was extracted by freeze-thaw cycles followed by phenol/ chloroform/isoamyl alcohol method. The single step PCR-RFLP assay was used to differentiate the assemblages between A and B, which were found in humans. In this method, 432 bp expected size was amplified, and then for detection of subspecies, specific restriction RsaI and BspLI enzymes were used.

Results

Totally 34 samples were positive in terms of Giardia cyst out of 720 examined samples microscopically, so the parasite spread rate is reported 4.72%. Analysis PCR-RFLP on these samples revealed that 28 samples (93.3%) have the genotype BIII and 2 samples (6.7%) belong to the subgroup BIV.

Conclusion

PCR-RFLP is a proper analytical method for determining the genotype among parasite types, using the glutamate dehydrogenizes zone’s genes. Based on the results, an animal origin of infection cycle is suggested.     

Sentinel-based Surveillance of Coyotes to Detect Bovine Tuberculosis, Michigan

Bovine tuberculosis (TB) is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of Michigan’s Lower Peninsula. Bovine TB in deer and cattle has created immense financial consequences for the livestock industry and hunting public. Surveillance identified coyotes (Canis latrans) as potential bio-accumulators of Mycobacterium bovis, a finding that generated interest in their potential to serve as sentinels for monitoring disease risk. We sampled 175 coyotes in the bovine TB–endemic area. Fifty-eight tested positive, and infection prevalence by county ranged from 19% to 52% (statistical mean 33%, SE 0.07). By contrast, prevalence in deer (n = 3,817) was lower (i.e., 1.49%; Mann-Whitney U_4,4 = 14, p<0.001). By focusing on coyotes rather than deer, we sampled 97% fewer individuals and increased the likelihood of detecting M. bovis by 40%. As a result of reduced sampling intensity, sentinel coyote surveys have the potential to be practical indicators of M. bovis presence in wildlife and livestock.     

PCR-RFLP Analysis of 28 SrDNA for Specification of Fasciola gigantica (Cobbold, 1855) in the Infected Lymnaea auricularia (Linnaeus, 1785) Snails from Northwestern Iran

Background

Fasciolosis in livestock is a crucial concern in the globe, mainly due to its impact on human health. The aim of this study was to determine the rate of infection with Fasciola gigantica (Cobbold, 1855) larvae in the field-collected snails of Lymnaea auricularia (Linnaeus, 1785) from northwestern Iran using a molecular approach.

Methods

A total of 6,759 pond snails were collected from 28 freshwater bodies in West Azarbaijan. PCR was performed to amplify a 618-bp fragment of the nuclear 28 SrRNA gene of Fasciola. The PCR products were digested by AvaII restriction enzyme to create restriction fragment length polymorphism (RFLP) patterns specific for the detection of F. gigantica.

Results

Of the total collected snails 496 (7.34 %) were L. auricularia, among which 4.64% (23 out of 496) were infected with a Fasciola species according to the PCR analysis. Only 2.22% (11 out of 496) of the infected snails were from the mountainous areas. The highest Fasciola infection rate recorded in the snails of a single site was 1.81% (9 out of 496 snails). Based on the RFLP pattern, F. gigantica accounted for 2.42% of the infection rates in the study sites.

Conclusion

Application of PCR-RFLP was proven to be a useful approach for valid and robust detection of the infection with F. gigantica in its intermediate host snails. These findings may therefore be applicable for establishment of the control programs against dissemination of the infection in different regions.