Biotyping of Multidrug Resistant <i>Campylobacter jejuni</i>from Poultry and Humans in Northern Region of Ghana

Thermophilic Campylobacters are zoonotic bacteria which are universally famous for causing enteritis in humans. They are normally found as commensals in the digestive tract of food animals with poultry being a major reservoir of the pathogenic species. This study highlighted the presence of Campylobacter in poultry (commercial and domestic) and humans (patients and asymptomatic individuals) and characterized strains by biotyping and susceptibility test in the Northern region of Ghana where animal husbandry is commonly practiced but zoonotic studies are limited. A total of 1087 specimens from stools of humans and cloacal swabs of poultry were screened from 25th October, 2017 to 7th May, 2019. Samples were cultured on modified charcoal-cefoperazone-deoxycholate agar and isolates identified using standard microbiological procedures and Lior Biotyping, while the antibiogram of isolates was determined by the Kirby-Bauer disk diffusion method. The frequency of Campylobacter was 43.1% and 12.9% respectively in poultry and humans. Campylobacter jejuni were recovered from 84% of commercial birds and 64% of domestic birds and in humans significantly fewer strains were observed in patients than asymptomatic individuals (p < 0.05). Biotype distribution revealed C. jejuni biotype I prevalence in domestic birds, patients and asymptomatic individuals whereas Bioytype II was largely found in commercial poultry. All isolated strains of Campylobacter were resistant to tetracycline and 69.4% of Campylobacter jejuni strains were resistant to erythromycin. Imipenem and the aminoglycosides were relatively effective as resistance of 10% and below 20% were respectively obtained. None of the endorsed treatment drugs (erythromycin, ciprofloxacin, and tetracycline) can be admitted in this region due to common resistance found among strains against these agents.

国产MALDI-TOF MS系统Smart MS 5020对临床微生物的鉴定性能评估

目的评估珠海迪尔Smart MS 5020型基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)系统及其2630种数据库(以下简称Smart MS 5020系统)对临床微生物的鉴定能力。方法共纳入2016—2020年浙江大学医学院附属第二医院留存的临床分离菌株913株,覆盖18个属52个种或复合群。采用直接涂抹法进行鉴定,比对仪器为德国布鲁克Biotyper MS系统及其DB2969数据库(以下简称Biotyper MS),对于两台质谱仪不一致的鉴定结果,采用16S rRNA扩增测序作为判定的金标准。针对复合群(鲍曼不动杆菌复合群、阴沟肠杆菌复合群以及肺炎克雷伯菌复合群),采用二代测序的鉴定结果作为金标准,统计学方法采用卡方检验。选取13株室内质控菌株作为重复性评估的检测样本,每株菌在Smart MS 5020系统上平行检测3次,判断其鉴定结果的一致性。结果对913株临床菌株,Biotyper MS系统种水平鉴定准确率为100%(913/913,95%CI:99.48%~99.58%),Smart MS 5020系统种水平鉴定准确率为99.78%(911/913,95%CI:99.12%~99.20%),2株表皮葡萄球菌鉴定为柯氏葡萄球菌;在临床微生物的鉴定能力方面,2种方法差异无统计学意义(P>0.05)。重复性评估结果显示,Smart MS 5020系统对13株室内质控菌株的鉴定结果完全一致。结论Smart MS 5020系统对临床常见微生物有很好的鉴定能力,包括复合群的鉴定能准确鉴定到复合群,对一些难区分菌如嗜血杆菌、链球菌等均能准确鉴定,但对复合群内具体种的细分能力有限,需要结合分子生物学、血清学或其他补充实验来准确鉴定。

Responses of Detoxifying, Antioxidant and Digestive Enzyme Activities to Host Shift of Bemisia tabaci (Hemiptera: Aleyrodidae)

The polyphagous B-biotype of Bemisia tabaci （Gennadius） has demonstrated a high capacity to adapt to numerous hosts from diverse plant families. To illustrate induced responses by the host plant, biochemical research on eight plant-insect interaction correlative enzymes, representing detoxifying, antioxidant and digestive pathways, were investigated. Transferring whitefly adults to Zhongmian 23 from the pre-adapted host Zhongmian 41 induced activities of carboxylesterase （by 1.80-fold）, glutathione S-transferase （by 3.79-fold）, proteinase （by 1.62-fold） and amylase （by 2.41-fold） activities, hut decreased polyphenol oxidase （by 1.89-fold） and peroxidase （by 1.88-fold）. However, transferring whitefly adults to the favorite host abutilon from Zhongmian 41 was associated with increased activities of cytochrome P450 monooxygenase （by 1.61-fold） and amylase （by 1.42-fold）, and decreased activities of polyphenol oxidase （by 2.96-fold） and peroxidase （by 2.07-fold）. Our results, together with previous studies, proved that multiple pathways are involved in coping with host shifts by polyphagous herbivores, and the taxonomic status and preference of the host transferred would affect which pathway would be induced. These results would represent a key challenge in developing long-term sustainable insect control strategies.

Sequence Analysis of mtDNA CO/Gene and Molecular Phylogeny of Different Geographical Populations of Bemisia tabaci (Gennadius)

Bemisia tabaci(Gennadius) is a serious pest in many cropping systems worldwide and occurs in different biotypes. ThemtDNA COI gene of the 12 Bemisia tabaci (Gennadius) populations from different regions and countries were analyzed.Based on mtDNA COI sequences, their biotypes were characterized and phylogenetic relationships among thesepopulations were established with the method of UPGMA. The results indicated the genetic similarity between thosepopulations from Beijing, Zhengzhou, Zaozhuang, Nanjing, Shanghai, Haikou, and the B-biotype populations fromCalifornia, Texas, Arizona reached 99.8-100%, which meant the nation-wide infested populations of B.tabaci in China inrecent years were B-biotypes. Another population collected from Kunming of Yunnan Province showed very high similaritywith Q-biotype B.tabaci from Spain and Morocco, which meant the Kunming population was Q-biotype. This is the firstreport on the invasion of Q-biotype into China.

cDNA Cloning of Heat Shock Protein Genes and Their Expression in an Indigenous Cryptic Species of the Whitefly Bemisia tabaci Complex from China

Thermal adaptation plays a fundamental role in shaping the distribution and abundance of insects,and heat shock proteins（Hsps）play important roles in the temperature adaptation of various organisms.To better understand the temperature tolerance of the indigenous ZHJ2-biotype of whitefly Bemisia tabaci species complex,we obtained complete cDNA sequences for hsp90,hsp70,and hsp20 and analyzed their expression profiles under different high temperature treatments by real-time quantitative polymerase chain reaction.The high temperature tolerance of B.tabaci ZHJ2-biotype was determined by survival rate after exposure to different high temperatures for 1 h.The results showed that after 41°C heat-shock treatment for 1 h,the survival rates of ZHJ2 adults declined significantly and the estimated temperature required to cause 50% mortality（LT50）is 42.85°C for 1 h.Temperatures for onset（Ton）or maximal（Tmax）induction of hsps expression in B.tabaci ZHJ2-biotype were 35 and 39°C（or 41°C）.Compared with previous studies,indigenous ZHJ2-biotype exhibits lower heat temperature stress tolerance and Ton（or Tmax）than the invasive B-biotype.

The biotypes and host shifts of cotton-melon aphids Aphis gossypii in northern China

Aphis gossypfi is a globally distributed species and therefore has a highly variable life cycle. Populations of A. gossypii in northern China exhibit greater genotypic diversity and a broader host range, yet the details of life cycles of different biotypes is still unclear. In this study, the Cytb and 16S gene regions of A. gossypfi collected from 5 common summer hosts and 4 primary hosts were analyzed. A total of 57 haplotypes were obtained from 1 046 individual A. gossyp# sequences. The sequence included 44 variable sites, 27 of which were parsimony informative sites and 17 of which were singleton variable sites. The most frequent 3 haplotypes were found in 896 individuals, representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual. A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals. Considering the individual host plant, 5 biotypes were identified. Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races. Type 3 was the most abundant biotype in cotton fields in northern China.

Comparative Analysis of Population Genetic Structure in Bemisia tabaci(Gennadius) Biotypes B and Q Based on ISSR Marker

Bemisia tabaci （Gennadius） biotypes B and Q are two invasive biotypes in the species complex. The comparison of the population genetic structure of the two biotypes is of significance to show their invasive mechanism and to their control. The intersimple sequence repeats （ISSR） marker was used to analyze the 16 B-biotype populations and 4 Q-biotype populations worldwide with a Trialeurodes vaporariorum population in Shanxi Province, China, and a B. tabaci non-B/Qbiotype population in Zhejiang Province, China, was used as control populations. The analysis of genetic diversity showed that the diversity indexes of biotype Q including Nei＇s gene diversity index, Shannon informative index, and the percentage of polymorphic loci were higher than those of biotype B. The high genetic diversity of biotype Q might provide the genetic basis for the excellent ecological adaptation. Cluster analysis suggested that the ISSR could not be used in the phylogenetic analysis though it could easily distinguish the biotypes of B. tabaci. The difference of the population genetic structure between the biotype B and the biotype Q exists based on the ISSR marker. Meanwhile, the results suggested that the molecular marker has its limitation in the phylogenetic analysis among the biotypes of B. tabaci.

Periodontally accelerated osteogenic orthodontics with platelet-rich fibrin in an adult patient with periodontal disease:A case report and review of literature

BACKGROUND In the clinical scenario,adult patients with periodontal diseases and dental malformation,characterized by dental crowding in lower anterior teeth with the thin biotype,often require orthodontic treatment.This case report aimed to evaluate the clinical and radiographic outcomes of periodontally accelerated osteogenic orthodontics(PAOO)combined with autologous platelet-rich fibrin(PRF)in an adult patient with class I malocclusion along with dental crowding,a thin periodontal biotype,and buccal plate deficiency.CASE SUMMARY A 32-year-old female complaining of dental crowding and gingival bleeding was referred to the orthodontic clinic.The patient underwent periodontal risk assessment prior to orthodontic treatment.She was diagnosed with a high risk of gingival recession due to dental crowding,root prominence,loss of buccal plates,and a thin gingival tissue biotype.The treatment regimen included PAOO combined with autologous PRF for alveolar augmentation and interproximal enamel reduction for moderate dental crowding.Clinically,PAOO-assisted orthodontic tooth movement in this case showed enhanced periodontium remodeling.Radiographic outcomes also showed statistically significant improvements(P<0.01)in the mandibular buccal alveolar bone.CONCLUSION This case report suggests the combination of autologous PRF with PAOO to enhance bone augmentation and long-term tissue support in adult orthodontic patients with periodontal disease.

Comparative Morphology and Morphometry of Six Biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae) from China

Morphology and morphometry of six biotypes （B, Q, Cv, ZHJ-1, ZHJ-2 and ZHJ-3） of Bemisia tabaci （Gennadius） from China on cotton plants were studied by using microscopes. Nymphal body sizes and characters were measured and observed, especially on the 4th instar （pupal case）, including the vasiform orifice, operculum, lingula, length and thickness of anterior and posterior wax margins, width of thoracic tracheal combs. Adult characters of both sexes were investigated including the body size, compound eyes, antennal segments, vasiform orifice, hind tarsi and genitals. The results indicated that differences of some morphological characters or morphometrics were significant among the six biotypes of B. tabaci in China： （1） Pupal sizes of the exotic biotypes （B and Q） were significantly larger than the indigenous biotypes with the following order as B〉Q〉ZHJ-I〉Cv〉ZHJ-3〉ZHJ-2; （2） for both male and female adults, sizes of all characters investigated in the invading biotypes （B and Q）, especially B, were much larger than those of the indigenous ones.

Studies on the Host Biotypes and Its Cause of Cotton Aphidin Nanjing,China

The fitness of cotton aphid, Aphis gossypii Glover, on different host plants was studied. The results showed that the cotton aphid population in Nanjing formed host preference bio-types on cucumber and cotton. The aphids from cotton and cucumber could not inter-transplant successfully between the two host plants. The apterous and alate offspring of stem mother from hibiscus could not survive, reproduce and form population normally when they were transplanted to cucumber and water melon, but they could form normally when transplanted to cotton and muskmelon. There was no significant difference between the stylet length of cotton aphids from cucumber and cotton, so the stylet length was not the cause of the host preference. When the aphids originated on hibiscus were transplanted to cotton, water melon and muskmelon, the stylet could reach host phloem to absorb host plant’s nutrients, but it couldn’t when transplanted to cucumber. Cotton aphid can feed on muskmelon without secretion of water-soluble saliv

Bemisia tabaci Biotype Dynamics and Resistance to Insecticides in Israel During the Years 2008-2010

The sweetpotato whitefly Bemisia tabaci（Hemiptera：Aleyrodidae）is an extremely polyphagous insect pest that causes significant crop losses in Israel and worldwide.B.tabaci is a species complex of which the B and Q biotypes are the most widespread and damaging worldwide.The change in biotype composition and resistance to insecticide in Israel was monitored during the years 2008-2010 to identify patterns in population dynamics that can be correlated with resistance outbreaks.The results show that B biotype populations dominate crops grown in open fields,while Q biotype populations gradually dominate crops grown in protected conditions such as greenhouses and nethouses,where resistance outbreaks usually develop after several insecticide applications.While in previous years,Q biotype populations were widely detected in many regions in Israel,significant domination of the B biotype across populations collected was observed during the year 2010,indicating the instability of the B.tabaci population from one year to another.Reasons for the changing dynamics and the shift in the relative abundance of B.tabaci biotype,and their resistance status,are discussed.

Isolation and Identification of Fusarium Wilt Pathogens from Bitter Gourd in Hainan Province

In order to isolate and identify the pathogens causing Fusarium wilt in bitter gourd in Hainan Province, China, four bitter gourd plants exhibiting symptoms of Fusarium wilt were collected from Tunchang County, Hainan Province. Four pathogen strains （HNTC-01, HNTC-02, HNTC-03 and HNTC-04） were isolated from the four plants, and identified via morphological observation and molecular techniques. The results showed that HNTC--01, HNTC-03 and HNTC-04 were F. oxysporum, while HNTC-02 was F. graminearum. The tests on pathogenicity and host specificity showed that HNTC-01, HNTC-03 and HNTC-04 were F. oxysporum Schl. f. sp. momordicae. Phylogenetic analysis revealed that HNTC-01, HNTC-03, HNTC-04 were clustered together with 14 strains of F. oxysporum, and HNTC-02 was clustered together with six strains of F. graminearum. In addition, rDNA-ITS region can be used to distinguish different Fusarium species, but can not be used to distinguish different biotypes within a Fusarium species. This study could provide a scientific basis for the diagnosis and prevention of wilt in bitter gourd.

An update on Gardneralla vaginalis associated bacterial vaginosis in Malaysia

Objective: To update the status of Gardnerella vaginalis(G. vaginalis) as a causative agent of bacterial vaginosis(BV) in Malaysia and to define its epidemiology, metronidazole resistance and virulence properties.Methods: It is a single-centre(Gynaecology clinic at the Hospital Kuala Lumpur,Malaysia) prospective study with laboratory-based microbiological follow up and analyses. Vaginal swabs collected from the patients suspected for BV were subjected to clinical BV diagnosis, isolation and identification of G. vaginalis, metronidazole susceptibility testing, vaginolysin and sialidase gene PCR, Piot's biotyping and amplified ribosomal DNA restriction analysis genotyping.Results: Among the 207 patients suspected for BV, G. vaginalis was isolated from 47 subjects. G. vaginalis coexisted with Trichomonas vaginalis and Candida albicans in 26 samples. Three G. vaginalis isolates were resistant to metronidazole. Biotyping revealed 1 and 7 as the common types. Amplified ribosomal DNA restriction analysis genotype II was found to be more common(n = 22; 46%) than I(n = 12; 25.53%) and III(n = 13;27.6%). All genotype I and III isolates carried the sialidase gene, while 91.6% and 84.6%contained the vaginolysin gene. Genotype I was significantly associated with postgynaecological surgical complications and abortions(P = 0.002).Conclusions: The existence of pathogenic G. vaginalis clones in Malaysia including drug resistant strains should not be taken lightly and needs to be monitored as these may bring more complications especially among women of child bearing age and pregnant women.

Nucleic Acid Determination and Risk Assessment of Brucella in Milk Products

Brucellosis is a worldwide zoonosis caused by Brucella, which poses great threat to human health. Totally 234 milk samples from a scale dairy farm were amplified by nested PCR, and the pathogens were further separated. The results showed that a DNA band of 419 bp was amplified from 18 out ot＂234 milk samples. Among 18 positive milk samples, nine samples amplified the DNA band of 535 bp, which were B. su/s ; one sample amplified the DNA bands of 535 bp and 285 bp, which was the mixture of B. su/s and B. bov/s. The results laid a foundation for prevention and control of brucellosis.

Species and Biotype Analysis of Brucella Strains Isolated from Cattle and Sheep in Xinjiang

[ Objective] The paper aimed to understand the epidemiological characteristics of Brucella strains in Xinjiang and then provide an available integrated measure to prevent and control brucellosis. [ Method ] Eleven suspected Brucella strains were isolated by traditional methods, which were further identified by AMOS-PCR assay. Conventional biochemical tests were carried out to identify the biological subtype of sheep Brucella. [ Result] Nine strains were all B. meliten- s/s, and biological test indicated that all of them were B. melitensis biotype 3. [ Conclusion] B. melitensis biotype 3 was the predominant strain of Brucella in Xin- jiang, and AMOS-PCR assay could be applied safely and quickly as an assistant tool to detect Brucella. The results of molecular epidemiology laid a foundation for updating prevention and control strategy against brucellosis in Xinjiang.

Impact of Rice Stem Borers and Identification of <i>Orseolia oryzivora</i>Harris &Gagné, 1982 (Diptera: Cecidomyiidae) Biotypes in the Southern Cameroon

The study compares the impact due to rice stem borers in two sites (Yaoundé and Ntui). It also shows the diversity of the African Rice Gall Midge (AfRGM) biotypes in southern Cameroon (Santchou, Ndop, Tonga, Ebolowa, Ba?gom, Yaoundé and Ntui). The New Rice for Africa (NERICA) varieties 3, 8, 9 and 13 sown in Ntui were less attacked than those sown in Yaoundé. At both sites, damages ranged from 0.78% to 2.7%. In terms of diversity, the main stem-borer species were O. oryzivora, Diopsis apicalis, D. longiconis and Chilo zacconius. Molecular analyses of Orseolia oryzivora larvae collected in the localities of Santchou, Ndop, Tonga, Ebolowa, Ba?gom and Yaoundé showed the existence of more than one O. oryzivora biotype in southern Cameroon’s rice basins.

Gingival biotype: The probe test utility

Purpose: To evaluate the relevance of the probe test in classifying the gingival biotype and to determine if differences exist between maxillary and mandibular gingival biotypes in the same patient. Materials and Methods: 53 subjects were included. The gingival thickness was evaluated as thick or thin. This evaluation was based on the transparency of the periodontal probe through the gingival margin while probing the sulcus. In another hand, 124 clinicians were asked to identify the gingival biotype in the same 53 subjects based on visual inspection on pictures. For each examiner, the accuracy in identifying the gingival biotype was assessed using the percentage of cases correctly identified. Results: the percentile of agreement is greater than 70% in maxilla as well as in mandible. An imbalance was observed on the maxillary pictures concerning the gingival biotype and on the thin biotype between maxilla and mandible despite there being no statistical difference (p < 0.0001). The percentile of agreement is quite similar in the mandible for both thin (74%) and thick (70%) gingival biotypes. Agreement between mandibular probe and maxilla probe results demonstrates a weak percentile of agreement of gingival biotypes between maxillary and mandibular anterior teeth (57%). Conclusion: The use of probe transparency through the gingival margin is an accurate and simple method for gingival biotype diagnosis to discriminate a thin gingival from a thick gingival. Furthermore, individualizing the gingival biotype for each of the arches appears to be a new outcome that needs further investigation.

Prevalence of Gingival Biotype in a Syrian Population and Its Relation to Tooth Shapes: A Cross-Sectional Study

Background: Gingival biotype is one of the most important factors that impede success in dental treatments;it affects the outcomes of periodontal surgery, and restorative treatments. Researchers classified gingival biotypes into thin biotype and thick biotype. It is crucial to identify tissue biotype before treatment. Aim of Study: To determine the prevalence of gingival biotypes in a Syrian population, in addition, to study the distribution of gingival biotypes according to gender and tooth shape. Material and Methods: This cross sectional study included 500 volunteers (300 males and 200 females) from the patients who had visited the department of periodontology-dental faculty at Damascus University. Gingival thickness was assessed to determine the gingival biotype for the maxillary central incisors using the direct measurement technique (Trans-gingival probing). Shapes of the maxillary incisors were recorded. A written informed consent was taken from each patient. Statistical analysis was done using test k2 p < 0.05. Results: The mean age was 26.8 ± 4.4 years. Thick gingival biotype was detected in 58.4% of the sample and most of patients are men while the prevalence of thin gingival biotype was 41.6% of the sample. It was also observed that patients with thin gingival biotype had triangular tooth shape in 99.5% (p < 0.05). Conclusion: Thick gingival biotype was observed to be more prevalent in a Syrian population than thin biotype. Most Syrian males had thick gingival biotype with square tooth shape while females had thinner biotype and triangular tooth shape.

Effect of Different Temperature and Humidity on Bemisia tabaci

Bemisia tabaci has many biotypes,and has become one of the main pests on vegetables,flowers,cotton and other crops,seriously influencing the safe production of crops.Temperature has a great influence on the survival,development,reproduction and behavior of Bemisia tabaci,which can affect the change of body temperature and metabolism of B.tabaci.It needs certain water to maintain normal life activities,such as digestion,nutrition transportation,and temperature regulation.There is a suitable humidity range for the growth and development of B.tabaci in different development stages.High humidity or drought have a great impact on the growth and development of B.tabaci,especially on its survival or reproduction.At the same time,the environmental temperature and humidity are the important factors that influence the geographical distribution and population change of B.tabaci.

Field Screening of Lesotho and South African Wheat Cultivars for Russian Wheat Aphid Resistance

Russian wheat aphid (Diuraphis noxia) is an international wheat pest and was first recorded in South Africa in 1978 in the Bethlehem area in the Eastern Free State. Le-sotho lies adjacent to one of the largest wheat producing areas in South Africa, the Eastern Free State, where winter wheat and facultative types are cultivated under dry land conditions. Wheat (Triticum aestivum L.) is an important crop adapted to all agro-ecological zones of Lesotho. Russian wheat aphid may have a significant impact on wheat yield. No monitoring or pest control is being done in Lesotho and at this stage there is very little information on the Russian wheat aphid resistance of wheat culti-vars cultivated in Lesotho. In view of this it is important to monitor the distribution of Russian wheat aphid biotypes in Lesotho and determine the level of Russian wheat aphid resistance in local Lesotho wheat cultivars. Two local Lesotho wheat cultivars, Bolane and Makalaote were screened together with South African cultivars Elands, Matlabas, Senqu, PAN3379, PAN3118 and SST387, in the glasshouse against all four known biotypes that occur in South Africa. All these cultivars were also planted in 5 m plots in the field at two localities Leribe and Roma in the lowlands of Lesotho. These cultivars were screened in the field for Russian wheat aphid resistance. The predomi-nant Russian wheat aphid biotypes in these areas were also determined. The Lesotho cultivar, Bolane had resistance against RWASA2 in the glasshouse, while Makalaote did not have any Russian wheat aphid resistance in either the glasshouse or field screenings. To contribute to food security an increasing wheat yield potential is a high priority. Russian wheat aphid has been included in the list of important international cereal pests. Russian wheat aphid adapts to changing environments and taking their ecology, distribution, virulence patterns, and variability into account is important in minimizing the gap between actual and attainable yields. Current management prac-tices for winter wheat in South Africa include the use of resistant cultivars, which is the most economical management strategy for Russian wheat aphid. Introducing Russian wheat aphid resistant cultivars in Lesotho will improve overall yield and as a result food security. This will also result in lower Russian wheat aphid pest pressure in the adjacent wheat production areas in the Eastern Free State, South Africa.