Phylo-geographic analysis of whitefly on the basis of mitochondrial cytochrome oxidase 1 gene

Abstract Bemisia tabaci is a species complex that causes damage to its broad range of plant hosts through serious feeding. It transmits plant viruses of different groups to important agricultural crops. Some important cash crops of Pakistan are sugar cane, rice, tobacco and seed oil. It shows high genetic variability and is differentiated as races or biotypes. Biotypes are, biotype Q, biotype B, biotype B2, biotype M, biotype L, biotype A, biotype H, biotype C, biotype K, biotype N, biotype R, biotype E, biotype P, biotype J, biotype S, biotype AN. Although the current report based on the Bayesian study of mitochondrial cytohrome oxidase gene1 (CO1) DNA sequences has classified the different populations of whiteflies into twelve genetic groups which are Mediterranean, Sub-Saharan Africa silverleafing, Indian Ocean, Asia II, Asia I, Australia, New World, Italy, China, Sub-Saharan Africa non-silverleafing, Mediterranean/Asia Minor/Africa and Uganda sweet potato. Begomoviruses is largest group of viruses transmitted by B. tabaci and cause major diseases of crops such as tomato and chili leaf curl disease, cassava mosaic disease; yellow mosaic disease of legumes and cotton leaf curl disease. The main objective of current study is to inculpate knowledge regarding genetic diversity of whitefly in cotton fields across Pakistan via analysis of partial DNA sequence of mitochondrial gene Cytochrom Oxidase I (mtCO1).

Resumo Bemisia tabaci é um complexo de espécies que causa danos a uma ampla gama de hospedeiros vegetais por meio de alimentação séria. Ele transmite vírus de plantas de diferentes grupos para importantes safras agrícolas. Algumas safras comerciais importantes do Paquistão são cana-de-açúcar, arroz, tabaco e óleo de semente. Apresenta alta variabilidade genética e é diferenciado em raças ou biótipos. Os biótipos são: biótipo Q, biótipo B, biótipo B2, biótipo M, biótipo L, biótipo A, biótipo H, biótipo C, biótipo K, biótipo N, biótipo R, biótipo E, biótipo P, biótipo J, biótipo S, biótipo AN . Embora o relatório atual baseado no estudo bayesiano das sequências de DNA do gene 1 da oxidase do citocromo mitocondrial (CO1) tenha classificado as diferentes populações de moscas-brancas em doze grupos genéticos, que são Mediterrâneo, África Subsaariana com folha de prata, Oceano Índico, Ásia II, Ásia I, Austrália, Novo Mundo, Itália, China, África Subsaariana sem folha prateada, Batata-doce Mediterrâneo / Ásia Menor / África e Uganda. Os begomovírus são o maior grupo de vírus transmitidos por B. tabaci e causam as principais doenças de culturas, como a doença do cacho do tomate e da pimenta-malagueta, doença do mosaico da mandioca, doença do mosaico amarelo de leguminosas e doença do enrolamento da folha do algodão. O principal objetivo do presente estudo é inculpar conhecimento sobre a diversidade genética da mosca-branca em campos de algodão em todo o Paquistão por meio da análise da sequência parcial de DNA do gene mitocondrial Citocromo Oxidase I (mtCO1).

Phylo-geographic analysis of whitefly on the basis of mitochondrial cytochrome oxidase 1 gene / Análise filogeográfica da mosca-branca com base no gene citocromo oxidase 1 mitocondrial

Bemisia tabaci is a species complex that causes damage to its broad range of plant hosts through serious feeding. It transmits plant viruses of different groups to important agricultural crops. Some important cash crops of Pakistan are sugar cane, rice, tobacco and seed oil. It shows high genetic variability and is differentiated as races or biotypes. Biotypes are, biotype Q, biotype B, biotype B2, biotype M, biotype L, biotype A, biotype H, biotype C, biotype K, biotype N, biotype R, biotype E, biotype P, biotype J, biotype S, biotype AN. Although the current report based on the Bayesian study of mitochondrial cytohrome oxidase gene1 (CO1) DNA sequences has classified the different populations of whiteflies into twelve genetic groups which are Mediterranean, Sub-Saharan Africa silverleafing, Indian Ocean, Asia II, Asia I, Australia, New World, Italy, China, Sub-Saharan Africa non-silverleafing, Mediterranean/Asia Minor/Africa and Uganda sweet potato. Begomoviruses is largest group of viruses transmitted by B. tabaci and cause major diseases of crops such as tomato and chili leaf curl disease, cassava mosaic disease; yellow mosaic disease of legumes and cotton leaf curl disease. The main objective of current study is to inculpate knowledge regarding genetic diversity of whitefly in cotton fields across Pakistan via analysis of partial DNA sequence of mitochondrial gene Cytochrom Oxidase I (mtCO1).

Bemisia tabaci é um complexo de espécies que causa danos a uma ampla gama de hospedeiros vegetais por meio de alimentação séria. Ele transmite vírus de plantas de diferentes grupos para importantes safras agrícolas. Algumas safras comerciais importantes do Paquistão são cana-de-açúcar, arroz, tabaco e óleo de semente. Apresenta alta variabilidade genética e é diferenciado em raças ou biótipos. Os biótipos são: biótipo Q, biótipo B, biótipo B2, biótipo M, biótipo L, biótipo A, biótipo H, biótipo C, biótipo K, biótipo N, biótipo R, biótipo E, biótipo P, biótipo J, biótipo S, biótipo AN . Embora o relatório atual baseado no estudo bayesiano das sequências de DNA do gene 1 da oxidase do citocromo mitocondrial (CO1) tenha classificado as diferentes populações de moscas-brancas em doze grupos genéticos, que são Mediterrâneo, África Subsaariana com folha de prata, Oceano Índico, Ásia II, Ásia I, Austrália, Novo Mundo, Itália, China, África Subsaariana sem folha prateada, Batata-doce Mediterrâneo / Ásia Menor / África e Uganda. Os begomovírus são o maior grupo de vírus transmitidos por B. tabaci e causam as principais doenças de culturas, como a doença do cacho do tomate e da pimenta-malagueta, doença do mosaico da mandioca, doença do mosaico amarelo de leguminosas e doença do enrolamento da folha do algodão. O principal objetivo do presente estudo é inculpar conhecimento sobre a diversidade genética da mosca-branca em campos de algodão em todo o Paquistão por meio da análise da sequência parcial de DNA do gene mitocondrial Citocromo Oxidase I (mtCO1).

Detection and characterization of Streptococcus downei, a rare bacterial species of mutans streptococci from caries-active patients

Background: The oral bacteria, mutans streptococci (MS), are an etiological agent of dental caries. Of MS, Streptococcus downei are rarely isolated bacteria. Aim: The aim of this study was to isolate and characterize S. downei from caries-active subjects. Materials and Methods: In all, 65 dental plaque samples were collected from dental caries-active subjects. All the isolates were further identified and characterized using 16S rDNA sequencing, biochemical tests, antibiogram, and minimum inhibitory concentration (MIC). Results: Five isolates have been identified as S. downei using 16S rDNA sequencing. Phylogenetic analysis showed that S. downei was closely related to S. sobrinus. The biotype traits of these five isolates were IV (n = 3), V (n = 1), and variants (n = 2). The study proposed one new biotype, classified as biotype VIII for the variant strain. The antibiogram tests revealed that all the strains of S. downei were susceptible to all the antibiotics used in the study with higher sensitivity to penicillin and ampicillin. The MIC of ampicillin and erythromycin against S. downei was 0.047 and 0.39 μg/mL, respectively. Conclusion: The study reports the prevalence of S. downei in caries-active subjects and recommends further investigations to determine its role in the disease.

Biotyping of Arcobacter butzleri isolated from poultry products: rescuing a phenotypic method / Biotipagem de Arcobacter butzleri desde produtos aviares: resgatando um método fenotípico

Arcobacter butzleri is an emergent zoonotic foodborne pathogen associated to enteritis and occasionally to bacteremia in human beings. Biotyping of this bacterium is important in order to establish the circulating strains and its dissemination routes. The purpose of this work was to determine the circulating A. butzleri biotypes in poultry products for human consumption in Southern Chile using the method proposed by Lior and Woodward, in order to explore the possibility of introducing this biotyping scheme as a routine laboratory tool. From the 60 strains studied the prevalent biotypes were 8A, 8B, 7A, 4A and 4B. The most frequently isolated biotype, independently of the sample of origin, was 8A with (44 strains, 73.3%). The less frequently isolated biotype was 4B (two strains 3.3%). The biotyping method used results to be simple, easy to handle and yields stable results. Therefore, it might be rescued to be used as a phenotypic tool for epidemiological marking of A. butzleri.

Arcobacter butzleri é um patógeno emergente, zoonótico e de transmissão alimentar, associado a enterite e, ocasionalmente, a bacteremia em seres humanos. A biotipagem desta bactéria é importante para estabelecer os biótipos circulantes e suas rotas de disseminação. Os objetivos deste trabalho foram determinar os biótipos de A. butzleri circulantes em alimentos de origem aviar para consumo humano, no sul do Chile, explorando a possibilidade de introduzir o método de biotipagem proposto por Lior e Woodward como uma ferramenta de rotina no laboratório. Entre as 60 cepas estudadas, os biótipos 8A, 8B, 7A e 4B foram os mais prevalentes. O biótipo mais frequentemente isolado, independentemente da amostra de origem, foi o biótipo 8A (44 cepas, 73,3%). O biótipo 4B apresentou a menor frequência de isolamento (duas cepas, 3,3%). O método de biotipagem utilizado resultou ser simples de executar, fácil de manusear e produz resultados estáveis. Portanto, pode ser resgatado para ser usado como uma ferramenta fenotípica para marcação epidemiológica de A. butzleri.

Biological typing and drug resistance analysis of Haemophilus strains from respiratory tract / 国际检验医学杂志

Objective To investigate season distribution,biological typing and drug resistant of Haemophitus in Qingdao Central Hospital.Methods The sputum and throat swab were collected from patients with respiratory tract infection,221 Haemophilus strains were identified and typed by the manual method and MicSCAN4 automatic analyzer,HNID identification plate.Antimicrobial susceptibility was tested by Kirby-Bauer method,and cephalosporins nitrate thiophene paper method was used to detect β-lacta-mase.Results A total of 96 strains of Haemophilus influenzae(1.6%)were isolated,10(10.4%)strains of Haemophilus influenzae were identified as type Ⅰ,31(32.3%)as type Ⅱ,40(41.7%)as typeⅢand 1 5(1 5.6%)as other types.A total of 125 strains Hae-mophitus parl influenzae(2.1%)were isolated,1 5 (12.0%)strains of Haemophilus parl influenzae were identified as type Ⅰ,23 (18.4%)as typeⅡ,69(55.2%)as type Ⅲ and 18(14.4%)as type Ⅳ,other types were not identified.The highest infected rate was in winter.Resistance rate of Haemophilus influenzae and Haemophitus parl influenzae to ampicillin were 40.6% and 44.8%,to tri-methoprim-sulfamethoxazole were 5 1.0% and 66.4%.The prevalence ofβ-lactamase of all strains were 40.6%and 44.8%.But sus-ceptible rates of Haemophilus to cefotaxime,cefuroxime,meropenem,chloramphenicol were over 90.0%.Conclusion The respira-tory tract infections to Haemophilus influenzae and Haemophitus parl influenzae is more frequently found in winter.Type Ⅱ and type Ⅲ are the most prevalent types.The resistance rates of Haemophilus to ampicillin and trimethoprim-sulfamethoxazole are in-creasing,should not be used as empirical treatment of Haemophilus infection.Antibiotics such as cefotaxime,cefuroxime,meropen-em could be chosen for the treatment of respiratory tract infection caused by Haemophilus.

Phenotypical characterization and adhesin identification in Escherichia coli strains isolated from dogs with urinary tract infections

Pathogenic strains of Escherichia coli are the most common bacteria associated with urinary tract infections in both humans and companion animals. Standard biochemical tests may be useful in demonstrating detailed phenotypical characteristics of these strains. Thirteen strains of E. coli isolated from dogs with UTIs were submitted to biochemical tests, serotyping for O and H antigens and antimicrobial resistance testing. Furthermore, the presence of papC, sfa, and afa genes was evaluated by PCR, and genetic relationships were established using enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). The antimicrobial that showed the highest resistance rate among the isolates was nalidixic acid (76.9%), followed by cephalotin (69.2%), sulfamethoxazole + trimethoprim (61.5%), tetracycline (61.5%), streptomycin (53.8%), ciprofloxacin (53.8%), ampicillin (46.2%), gentamicin (30.8%) and chloramphenicol (23.1%). No isolate was resistant either to meropenem or nitrofurantoin. Among the five clusters that were identified using ERIC-PCR, one cluster (A) had only one strain, which belonged to a serotype with zoonotic potential (O6:H31) and showed the genes papC+, sfa+, afa-. Strains with the genes papC-, sfa+, afa- were found in two other clusters (C and D), whereas all strains in clusters B and E possessed papC-, sfa-, afa- genes. Sucrose and raffinose phenotypic tests showed some ability in discriminating clusters A, B and C from clusters D and E.

Biotypes and Enterotoxigenicity of Staphylococci Isolated from Camel’s Meat in Jordan.

A total of 264 camel’s meat and nasal swab samples were collected for isolation and typing of Staphylococci from Irbid Governorate in northern Jordan. About 97 % and 85% of meat and nasal swabs samples showed typical colonies of Staphylococcus aureus on Baird- Parker agar respectively. Out of 243 presumptively identified isolates, only 74 and 64 were confirmed as S. aureus by Microbact system and PCR technique respectively. About 67% of the isolates were typable by Devriese’s scheme. Fifteen of those isolates (23%) were specifically allocated to human, bovine, ovine or abattoir where, 14% of these host specific isolates belonged to human biovar. The other 44% belonged to non-host specific biovars with majority of them were allocated to NHS1 biovar. When tested for the presence of toxin genes, 71.9% of S. aureus isolates had SE(s) genes with SEA being the most prominent at 91.3%. The study also showed that not only coagulase positive isolates contain toxin genes, coagulase negative isolates also possess toxin genes and thus are considered potential hazards in camel’s meat.

Differentiation of Candida dubliniensis from Candida albicans with the use of killer toxins / Avaliação das toxinas killer na diferenciação entre Candida albicans e Candida dubliniensis

The aim of this study was to report the ability of killer toxins, previously used as biotyping techniques, as a new tool to differentiate C. albicans from C. dubliniensis. The susceptibility of C. albicans and C. dubliniensis to killer toxins ranged from 33.9 to 93.3 percent and from 6.67 to 93.3 percent, respectively.

Avaliou-se a capacidade das toxinas killer, previamente utilizadas na biotipagem de C. albicans, como método para diferenciar C. albicans de C. dubliniensis. A susceptibilidade de C. albicans e C. dubliniensis às toxinas killer variou de 33,9 por cento a 93,3 por cento para C. albicans e de 6,67 por cento a 93,3 por cento para C. dubliniensis.

Phenotypic Characterization of Salmonella Typhimurium Isolates from Food-animals and Abattoir Drains in Buea, Cameroon.

Salmonella spp. have been extensively incriminated worldwide as common causes of bacterial gastroenteritis in humans, with food-animals serving as important reservoirs. The study was aimed at investigating cattle and pigs slaughtered in Buea as reservoirs of Salmonella Typhimurium and the susceptibility of isolates to antibiotics. In total, 230 specimens (comprising 50 each from the rectum, ileum, and gall bladder of cattle; and 10 each from same anatomical sites of pigs and 50 from abattoir drains) were analyzed for Salmonella using the standard microbiological, biochemical and serological techniques. Antibiotic susceptibility of the isolates was determined by the Kirby-Bauer disc-diffusion test. The isolates were characterized into biotypes using the API 20E kit, and results were analyzed using the chi-square test. Seventy-five (32.6%) of the 230 specimens were positive for S. Typhimurium, with pigs and abattoir drains presenting the highest level of isolation (40%). Biochemical typing grouped the isolates into five biotypes. Biotype I was the most prevalent (30.6%) while biotype IV was the least prevalent (9.3%) and was absent in samples from pigs. Antibiotic susceptibility studies revealed 14 antibiotypes based on antibiotics used in the study. The predominant antibiotype AMXR DOXRCEFR was recorded in 13 (17.3%) of the isolates. Multidrug resistance (to four or more antibiotics) was recorded in 50.7% (38/75) of the isolates. The most active drugs were ciprofloxacin (98.6%), ofloxacin (93.3%), amikacin (90.6%), and gentamicin (84%). All the isolates (100%) were resistant to tetracycline and ampicillin. Cattle and pigs were found to be reservoirs of S. Typhimurium in the environment of Buea, Cameroon, implying that foods from these sources, if not properly handled, could serve as vehicles for its transmission to humans.

Study on characteristics of Haemophilus influenzae strains by phenotyping and pulsed-field gel electrophoresis genotyping / 中华微生物学和免疫学杂志

objective To investigate the epidemiological and molecular typing features of the pathogenic Haemophilus influenzae(H.influenzae)by biotyping,serotyping and pulsed-field gel electrophoresis(PFGE).Methods A total of 273 invasive isolates of H influenzae were collected from the pediatric patients with pneumonia at Chengdu Children Hospital of Sichuan province from 1988 and 2004 to 2007.The idenbfication of H.influenzae strains were done according to the laboratory standard methodology described by Manual of Clinical Microbiology(American).All strains were biotyped according to Kilian's classification with the API[R]NH system.And serotyped by a slide agglutination assay with type a to f specific antlaerum as described by Pittman.PCR method for identification of H.influenzae were performed as described by Falla.One hundred of 273 strains were analyzed by PFGE as described by Saito with some modifications.The resuIts of PFGE were analyzed by Bionumerics soft(Version 4.0,Applied Maths BVBA,Belium).Restilts 78.2%of 273 cases occurred under 1 years old.Eight biotypes were found among the 273 H.influenzae isolates.17.6%(48/273)of all isolates belonged to biotype Ⅰ,43.6%(119/273)were biotype Ⅱ,22.7%(62/273)were biotype Ⅲ,7.3%(20/273)were biotype Ⅳ,5.9%(16/273)were biotype Ⅴ,0.4%(1/273)were biotype Ⅵ,1.8%(5/273)were biotype Ⅶ and 0.7%(2/273)were biotype Ⅷ.respeetively.99.6% of all 273 isolates were nontypeable.There was only one isolate was serotvpe f Ninty-six PFGE genotypes were obtained in this study.One hundred strains demonstrated a variety of genomic Datterns by PFGE.The most isolates of the flame PFGE genotype(type 35)was 3 isolates.Each of93 PFGE genotypes was represented by only a single isolate.The genotypes distribution didn't correlate with the time distribution of the strains were isolated.Conclusion Nontypeable H.influenzae primarily caused acute Dneumoma in children under 1 years old.They mostly belonged to biotype Ⅰ,Ⅱ and Ⅲ biotypes.The nontypeable H.influenzae strains appeared to more heterogeneous patterns by PFGE genotyping.Genotyping may helP understand the molecular characteristics of outbreak and endemicity according to the results of PFGE.PFGE genotyping proved to have a much stronger discriminatory power than either serotyping or biotyping.Our findings suggest that PFGE analysis is useful for the epidemiologieal study of H.influenzae infections.

Phenotypic characterization of Candida spp. isolates from chronic periodontitis patients

Aim: Several typing methods for Candida spp. have been suggested in the literature in order to distinguish isolates for studies about the virulence or infection routes of these microorganisms and, in particular, for epidemiological purposes. The aim of this study was to establish a comparison between the phenotypic profile of oral Candida isolates from periodontitis patients and control individuals. Methods: The morphotyping and biotyping of 35 C. albicans isolates obtained from chronic periodontitis patients and 48 isolates from control individuals were performed. For morphotyping, the isolates were plated on malt extract agar and incubated for 10 days. Sixteen different morphotypes were observed for C. albicans, the most frequently observed being 0000 and 0001. Results: Biotype 0000 (complete absence of fringe) was most prevalent among the isolates obtained from periodontitis patients compared to those from control individuals, with statistical significance. Biotyping revealed 5 different biotypes with higher prevalence of the biotype 357 among the isolates from control and periodontitis groups. Conclusions: The results obtained by biotyping of the isolates did not permit to differentiate a characteristic model related to periodontal disease, whilst the morphotype 0000 was most frequently isolated from periodontitis patients.

Typing of Acinetobacter baumannii isolated from patients in intensive care unit with IRS-PCR / 第三军医大学学报

Objective To type Acinetobacter baumannii using infrequent restriction site PCR (IRS PCR). Methods Strain specific electrophoretic patterns from PCR products by amplifying DNA sequences flanking infrequent restriction sites of 15 strains of Acinetobacter baumannii were compared with the results of biotyping and antimicrobial susceptibility. Results The 15 bacteria were divided into 5 gene types with IRS PCR, but 3 with biotyping, and 4 with antimicrobial susceptibility. Conclusion IRS PCR method for typing Acinetobacter baumannii is of strong sensitivity, high recognition, good repeatability, convenient operation, and wide range of application.

Characteristics of Shigella sonnei Isolated in Kyungsangbuk-do in 1998: Biochemical Characteristics, Biotyping, Antibiotic Resistance Pattern, and Plasmid Profile / 한국역학회지

Thirty nine strains and 109 strains of Shigella sonnei were isolated from the outbreaks of Youngchun and Kyungju, respectively, while 15 strains were isolated from sporadic cases of eight regions in Kyungbuk province from September to November in 1998. We investigated the relationship among the S. sonnei strains by using biochemical characteristics, biotyping, antibiotic resistance pattern, and plasmid profile. Among the isolates, seven strains of S. sonnei isolated in Youngchun showed gelatin hydrolyase positive but the others showed gelatin hydrolyase negative. One hundred and fifty two strains were a type, while eleven among thirty nine strains isolated in Youngchun were g type. Antibiotics resistance patterns of S. sonnei strains isolated in Youngchun and Kyungju were significantly different. Thirty nine strains of S. sonnei isolated in Youngchun were resistant to SM, TE, and TMP/SMX, while eighty six of S. sonnei among one hundred and nine strains isolated in Kyungju were resistant to AM, CB, K, SM, TE, and TMP/SMX. Antibiotics resistance patterns of residual twenty three isolates were similar to those of eighty six strains. The Plasmid profiles of strains of S. sonnei isolated from the Kyungju were different from those of S. sonnei strains isolated in Youngchun. The Plasmid profiles of S. sonnei strains isolated from Youngchun were identical to those of a S. sonnei strains randomly selected from the outbreak in Daegu in 1998. The Plasmid profiles of S. sonnei strains isolated from Kyungju were identical to those of two strains of S. sonnei randomly selected from the outbreaks of Kanglung and Wonju in 1998. From the above results, it is considered that the strains of S. sonnei isolated from Kyungju and Youngchun region are not identical clone.