Isolation and characterization of bacteria associated with silkworm gut under antibiotic-treated larval feeding / Isolamento e caracterização de bactérias associadas ao trato intestinal do bicho-da-seda sob alimentação larval tratada com antibióticos

Abstract The impact of antibiotics on growth, cocoon production was assessed in addition to isolation and characterization of bacteria associated with silkworm gut of infected larvae. Larval rearing was maintained at recommended conditions of temperature and humidity. Silkworm larvae showing abnormal symptoms were collected from the control group and dissected for gut collection. Bacteria were isolated from the gut content by spreading on agar plates and incubated at 37 °C for 48 hrs. Bacterial identification and phylogenetic analysis were carried out by 16S rRNA gene sequencing. The isolated bacteria were subjected to antimicrobial susceptibility test (disc diffusion methods) by using Penicillin (10 µg/mL), Tetracycline (30 µg/mL), Amoxicillin (25 µg/mL), Ampicillin (10 µg/mL), and Erythromycin (15 µg/mL). All isolated strains showed positive results for the catalase test. We isolated and identified bacterial strains (n = 06) from the gut of healthy and diseased silkworm larvae. Based on the 16S rRNA gene sequence, isolated bacteria showed close relation with Serratia, Bacillus, and Pseudomonas spp. Notably, 83.3% of strains were resistant to Penicillin, Tetracycline, Amoxicillin, Ampicillin, and Erythromycin but 16.6% showed antibiotic susceptibility to the above-mentioned commonly used antibiotics. Silkworm larvae fed on penicillin-treated leaves showed significant improvement in larval weight, larval length, and cocoon production. Significantly higher larval weight (6.88g), larval length (5.84cm), and cocoon weight (1.33g) were recorded for larvae fed on leaves treated with penicillin as compared to other antibiotics. Isolated bacterial strains showed close relation with Serratia spp., Bacillus spp. and Pseudomonas spp.

Resumo O impacto dos antibióticos no crescimento e na produção do casulo foi avaliado, além do isolamento e caracterização das bactérias associadas ao intestino de larvas infectadas do bicho-da-seda. A criação das larvas foi mantida nas condições recomendadas de temperatura e umidade. As larvas do bicho-da-seda com sintomas anormais foram coletadas do grupo controle e dissecadas para coleta do intestino. As bactérias foram isoladas do conteúdo intestinal por espalhamento em placas de ágar e incubadas a 37° C durante 48 horas. A identificação bacteriana e a análise filogenética foram realizadas pelo sequenciamento do gene 16S rRNA. As bactérias isoladas foram submetidas a teste de sensibilidade antimicrobiana (métodos de difusão em disco) com penicilina (10 µg / mL), tetraciclina (30 µg / mL), amoxicilina (25 µg / mL), ampicilina (10 µg / mL) e eritromicina (15 µg / mL). Todas as cepas isoladas apresentaram resultados positivos para o teste da catalase. Isolamos e identificamos cepas bacterianas (n = 06) do intestino de larvas de bicho-da-seda saudáveis e doentes. Com base na sequência do gene 16S rRNA, as bactérias isoladas mostraram estreita relação com Serratia, Bacillus e Pseudomonas spp. Notavelmente, 83,3% das cepas eram resistentes a penicilina, tetraciclina, amoxicilina, ampicilina e eritromicina, mas 16,6% mostraram suscetibilidade aos antibióticos comumente usados mencionados acima. As larvas do bicho-da-seda alimentadas com folhas tratadas com penicilina apresentaram melhora significativa no peso larval, comprimento larval e produção de casulo. Peso larval significativamente maior (6,88g), comprimento larval (5,84cm) e peso do casulo (1,33g) foram registrados para larvas alimentadas com folhas tratadas com penicilina, em comparação com outros antibióticos. Cepas bacterianas isoladas mostraram estreita relação com Serratia spp., Bacillus spp. e Pseudomonas spp.

Screening, biochemical characterization and antibiotics resistance/susceptibility of bacteria isolated from native soil and water samples / Triagem, caracterização bioquímica e resistência/suscetibilidade a antibióticos de bactérias isoladas de amostras nativas de solo e água

The present study was conducted to isolate and characterize bacteria from water and soil sample taken from the Lahore Canal at different sites i.e. Mall Road, Mohlanwal and Khera site. Isolated bacterial strains were identified on the basis of morphological and biochemical tests. Identification was confirmed by culturing bacteria on selective media. Antibiotic resistance test was also performed to observe the resistance of bacteria against different antibiotics. Blood agar test was performed for identification of different pathogenic bacteria. The result revealed that water and soil samples of Lahore Canal Lahore from different sites were contaminated with Escherichia coli, Salmonella sp., Vibrio sp., Bacillus spp., Enterococcus sp. and Staphylococcus spp. Due to presence of these pathogens, this water is not suitable for any domestic and irrigation use. Study also revealed that water of the Lahore Canal is harmful for human health as it is contaminated with bacteria that can cause severe disease e.g., Escherichia coli can cause gastroenteritis, Bacillus spp. can cause nausea and vomiting, Enterococcus may infect urinary tract, Salmonella sp. is responsible for Bacteremia, Staphylococcus spp. can cause mild fever and Vibrio sp. can be the reason of cholera. Thus it is rendered unfit for any kind of human use even other than drinking like swimming, bathing, washing etc., until and unless some remedial measures are employed to eradicate pathogenic microorganisms by WASA and LWMS according to standards of WHO. Similarly, it is quite harmful, when and where ever it is used for irrigation without proper treatment.

O presente estudo foi realizado para isolar e caracterizar bactérias de amostras de água e solo retiradas do Canal Lahore, em Lahore, em diferentes locais, ou seja, Mall Road, Mohlanwal e Khera. As cepas bacterianas isoladas foram identificadas com base em testes morfológicos e bioquímicos. A identificação foi confirmada por cultura de bactérias em testes de meios seletivos. O teste de resistência aos antibióticos também foi realizado para observar a resistência das bactérias a diferentes antibióticos. Foi realizado o teste de ágar sangue para identificar diferentes bactérias patogênicas. O resultado revelou que amostras de água e solo do Canal Lahore, Lahore, de diferentes localidades estavam contaminadas com Escherichia coli, Salmonella sp., Vibrio sp., Bacillus spp., Enterococcus sp. e Staphylococcus spp. Por causa da presença desses patógenos, essa água não é adequada para qualquer uso doméstico e de irrigação. O estudo revelou que a água do Canal Lahore é prejudicial à saúde humana, pois está contaminada com bactérias que podem causar doenças graves, por exemplo: Escherichia coli pode ocasionar gastroenterite; Bacillus spp. pode causar náuseas e vômitos; Enterococcus sp. pode infectar o trato urinário; Salmonella sp. é responsável pela bacteremia; Staphylococcus spp. pode causar febre leve; e Vibrio sp. pode ser a razão da cólera. Assim, torna-se imprópria para uso humano, como natação, banho, lavagem etc., até que algumas medidas corretivas sejam empregadas para erradicar microrganismos patogênicos por WASA e LWMS de acordo com os padrões da OMS. Da mesma forma, é bastante prejudicial, quando usada para irrigação sem tratamento adequado.

Isolation and characterization of bacteria associated with silkworm gut under antibiotic-treated larval feeding

Abstract The impact of antibiotics on growth, cocoon production was assessed in addition to isolation and characterization of bacteria associated with silkworm gut of infected larvae. Larval rearing was maintained at recommended conditions of temperature and humidity. Silkworm larvae showing abnormal symptoms were collected from the control group and dissected for gut collection. Bacteria were isolated from the gut content by spreading on agar plates and incubated at 37 °C for 48 hrs. Bacterial identification and phylogenetic analysis were carried out by 16S rRNA gene sequencing. The isolated bacteria were subjected to antimicrobial susceptibility test (disc diffusion methods) by using Penicillin (10 µg/mL), Tetracycline (30 µg/mL), Amoxicillin (25 µg/mL), Ampicillin (10 µg/mL), and Erythromycin (15 µg/mL). All isolated strains showed positive results for the catalase test. We isolated and identified bacterial strains (n = 06) from the gut of healthy and diseased silkworm larvae. Based on the 16S rRNA gene sequence, isolated bacteria showed close relation with Serratia, Bacillus, and Pseudomonas spp. Notably, 83.3% of strains were resistant to Penicillin, Tetracycline, Amoxicillin, Ampicillin, and Erythromycin but 16.6% showed antibiotic susceptibility to the above-mentioned commonly used antibiotics. Silkworm larvae fed on penicillin-treated leaves showed significant improvement in larval weight, larval length, and cocoon production. Significantly higher larval weight (6.88g), larval length (5.84cm), and cocoon weight (1.33g) were recorded for larvae fed on leaves treated with penicillin as compared to other antibiotics. Isolated bacterial strains showed close relation with Serratia spp., Bacillus spp. and Pseudomonas spp.

Resumo O impacto dos antibióticos no crescimento e na produção do casulo foi avaliado, além do isolamento e caracterização das bactérias associadas ao intestino de larvas infectadas do bicho-da-seda. A criação das larvas foi mantida nas condições recomendadas de temperatura e umidade. As larvas do bicho-da-seda com sintomas anormais foram coletadas do grupo controle e dissecadas para coleta do intestino. As bactérias foram isoladas do conteúdo intestinal por espalhamento em placas de ágar e incubadas a 37° C durante 48 horas. A identificação bacteriana e a análise filogenética foram realizadas pelo sequenciamento do gene 16S rRNA. As bactérias isoladas foram submetidas a teste de sensibilidade antimicrobiana (métodos de difusão em disco) com penicilina (10 µg / mL), tetraciclina (30 µg / mL), amoxicilina (25 µg / mL), ampicilina (10 µg / mL) e eritromicina (15 µg / mL). Todas as cepas isoladas apresentaram resultados positivos para o teste da catalase. Isolamos e identificamos cepas bacterianas (n = 06) do intestino de larvas de bicho-da-seda saudáveis e doentes. Com base na sequência do gene 16S rRNA, as bactérias isoladas mostraram estreita relação com Serratia, Bacillus e Pseudomonas spp. Notavelmente, 83,3% das cepas eram resistentes a penicilina, tetraciclina, amoxicilina, ampicilina e eritromicina, mas 16,6% mostraram suscetibilidade aos antibióticos comumente usados mencionados acima. As larvas do bicho-da-seda alimentadas com folhas tratadas com penicilina apresentaram melhora significativa no peso larval, comprimento larval e produção de casulo. Peso larval significativamente maior (6,88g), comprimento larval (5,84cm) e peso do casulo (1,33g) foram registrados para larvas alimentadas com folhas tratadas com penicilina, em comparação com outros antibióticos. Cepas bacterianas isoladas mostraram estreita relação com Serratia spp., Bacillus spp. e Pseudomonas spp.

Antibacterial activity of Ricinus communis plant extract against antibiotic resistant Helicobacter pylori and Gluconobacter oxydans isolated from fresh apple juices samples

Abstract Bacteria were isolated from samples of Fresh Apple juices from shops of three different localities of Lahore. Analysis of samples from Liberty, Anarkali and Yateem khana Markets show different levels of contamination. There were pathogenic and non-pathogenic bacteria in all samples and were identified by the morphological and biochemical tests. Most of the plasmids of pathogenic bacteria were 4kb in their molecular size. Ribotyping of 16S ribosomal RNA gene sequencing was done to confirm Helicobacter pylori strain and Gluconobacter oxydans. The highest sensitivity of 210mm was shown by Enterobacter sp. against Aztheromysine disk (15µg) while Micrococcus sp. was highly resistant against all of the Antibiotics applied. The antibiotic resistance of pathogenic bacteria was also checked against Ricinus communis plant's extracts, all isolated bacterial pathogens were resistant but only, E.coli was inhibited at 300µl of the extracts. Presence of pathogenic bacteria in Apple juice samples was due to contamination of sewage water in drinking water while some of these pathogenic bacteria came from Apple's tree and other from store houses of fruits.

Resumo As bactérias foram isoladas de amostras de suco de maçã fresco de lojas de três diferentes localidades de Lahore. A análise de amostras dos mercados Liberty, Anarkali e Yateem khana mostram diferentes níveis de contaminação. Havia bactérias patogênicas e não patogênicas em todas as amostras e foram identificadas pelos testes morfológicos e bioquímicos. A maioria dos plasmídeos de bactérias patogênicas tinha 4 kb em seu tamanho molecular. A ribotipagem do sequenciamento do gene do RNA ribossômico 16S foi realizada para confirmar a cepa de Helicobacter pylori e Gluconobacter oxydans. A maior sensibilidade de 210 mm foi mostrada por Enterobacter sp. contra disco de azteromisina (15µg) enquanto Micrococcus sp. foi altamente resistente a todos os antibióticos aplicados. A resistência a antibióticos de bactérias patogênicas também foi verificada contra extratos de plantas de Ricinus communis, todos os patógenos bacterianos isolados foram resistentes, mas apenas E. coli foi inibida em 300µl dos extratos. A presença de bactérias patogênicas nas amostras de suco de maçã deveu-se à contaminação da água de esgoto na água potável, enquanto algumas dessas bactérias patogênicas vieram da árvore da maçã e outras de armazéns de frutas.

Unlocking the molecular realm: advanced approaches for identifying clinically and environmentally relevant bacteria

Rapid, effective, and specific identification of clinical and environmental bacterial pathogens is of major importance for their control. Traditionally, bacteria have been identified by phenotypic methods based on morphological, biochemical, and metabolic properties. While these methods are very useful in clinical practice, they have limitations including a poor ability to differentiate within and between species and time-consuming workflows. Newly developed molecular methods can greatly improve the accuracy of taxonomic characterization, identifying specific strains of medical or environmental importance. However, due to high costs and the need for trained professionals, these methods are not yet routine in diagnostic laboratories. Thus, disseminating knowledge on advances in molecular identification techniques is pivotal to make these methodologies accessible. The objective of this work was to review and discuss current molecular techniques for bacteria identification aiming to track and monitor microbial agents in clinical and environmental samples.

Spectrum of bacterial keratitis in North India: A retrospective analysis of six years at a tertiary care eye center

Purpose: To analyze the pattern of bacterial pathogens causing infective keratitis and their resistance to the recommended antibiotics over six years. Methods: It was a retrospective study of 9,357 cases of bacterial keratitis from January 2015 to December 2020, at a tertiary care ophthalmic center. A total of 9,547 corneal specimens were obtained from the study subjects. Demographic details of the patients, pathogenic bacteria isolated, and their antimicrobial susceptibility were noted and analyzed. Results: Bacterial pathogens were identified in 23.52% of the specimens. The most common isolates were coagulase?negative Staphylococci (60.75%), followed by Pseudomonas aeruginosa (14.23%), Staphylococcus aureus (13.92%), gram negative bacilli of the family Enterobacterales (8.64%), Streptococcus spp. (1.72%), Acinetobacter spp. (0.13%), and other non?fermenting gram?negative bacilli (0.57%). In Staphylococci, 55–80% of isolates were resistant to erythromycin, and 40–70% to fluoroquinolones, while no resistance was observed against vancomycin. 40–60% of isolates of P. aeruginosa were resistant to cephalosporins, 40–55% to fluoroquinolones, and 30–60% to aminoglycosides. Also, 40–80% of isolates of Enterobacterales were resistant to cephalosporins, and 50–60% to fluoroquinolones. Most gram?negative isolates were susceptible to carbapenems and polymyxin B. Conclusion: To the best of our knowledge, our study is the largest compilation of microbiological profile of bacterial keratitis from North India. It highlights the current trend of the bacterial pathogens that cause infectious keratitis. Staphylococci and Pseudomonas were found to be the most common pathogens. Increased resistance was seen against some of the commonly prescribed empirical antibiotics. Such evidence is useful for restructuring the empirical prescription practices from time to time.

Isolation and characterization of aerobic actinomycetes with probiotic properties in Nile tilapia

This study scoped the isolation of aerobic actinomycetes with probiotic properties against bacterial pathogens in Niletilapia. Eleven rhizosphere soil samples were collected from the agricultural sites in three provinces (Chanthaburi,Nan, and Chachoengsao) of Thailand. A total of 157 actinomycete-like colonies were successfully isolated. Theantibacterial testing against four tested bacteria (Streptococcus agalactiae 2809, Aeromonas jandaei 1929, Aeromonasveronii 1930, and Edwardsiella ictaluri 2234) was carried out by a modified cross-streaked method. The results showedthat 108 strains possessed antibacterial activity against at least one of the bacterial pathogens. Seventeen active isolateswere identified in the actinomycetes by the analysis of the partial 16S ribosomal rRNA gene, and the phylogeneticrelationships of the isolates and their closely related strains were confirmed by the neighbor-joining method. IsolatesLNW002 and YNW004 survived in the liquid cultivation with International Streptomyces Project 2 at pH 2 and thepresence of 0.3% bile salt for 2 hours, which mimics the gastric acidity and bile salt in the gastrointestinal tract of Niletilapia in vitro. In conclusion, these strains might be further investigated for their efficacy as probiotics in Nile tilapia.This study is the first to report on anti-E. ictaluri activity in streptomycetes.

Assessment and incidence of fish associated bacterial pathogens at hatcheries of Azad Kashmir, Pakistan / Avaliação e incidência de patógenos bacterianos associados a peixes em incubadoras da Caxemira de Azad, Paquistão

Abstract Fish is the most indispensable source of proteins for individuals and have high nutritional value. On the other hand, the fish culturing raised issues of fish health due to close contact between the aquatic environment and the fish pathogens. So, the aim of the current study was to identify the bacterial pathogens and screen the injured Rainbow trout rearing in different trout hatcheries run under fisheries department of the government of Azad Jammu and Kashmir, Pakistan. Seven bacterial pathogens such as Shigella flexneri, Enterobacter amnigenus, Salmonella Typhimurium, Serratia odorifera, Pseudomonas aeruginosa, Streptococcus pyogenes, and Bacillus cereus were isolated and identified. Results revealed that the injury of fish specimens was due to overcrowding. Instead of rainbow coloration, specimens have darker black in color. The water of ponds was not clean and clear and such conditions was because of the greater quantity of feed thrown in the water. It was concluded that poor hygienic water condition and overloading allowed the opportunistic bacterial contaminations to succeed which cause a serious threat to hatcheries.

Resumo O peixe é a fonte mais indispensável de proteínas para os indivíduos e tem alto valor nutricional. Por outro lado, a cultura dos peixes levantou questões sobre a saúde dos peixes devido ao próximo contato entre o ambiente aquático e os agentes patogênicos desses peixes. Assim, o objetivo do presente estudo foi identificar os patógenos bacterianos e rastrear a criação da truta arco-íris que apresentou lesões em diferentes incubadoras de trutas, com supervisão do departamento de pesca do governo de Azad Jammu e Caxemira, Paquistão. Sete patógenos bacterianos foram isolados e identificados, tais como: Shigella flexneri, Enterobacter amnigenus, Salmonella typhimurium, Serratia odorifera, Pseudomonas aeruginosa, Streptococcus pyogenes e Bacillus cereus. Os resultados revelaram que a lesão de espécimes de peixes foi devido à superlotação. Em vez da coloração do arco-íris, os espécimes tiveram uma coloração preta mais escura. A água das lagoas não era limpa e nem clara, e tais condições ocorreram devido a maior quantidade de alimento lançada na água. Concluiu-se que a precária condição higiênica da água e também a sobrecarga permitiram que as contaminações bacterianas oportunistas fossem bem-sucedidas, causando séria ameaça às incubadoras.

Antibacterial Activity of Glycyrrhiza Glabra Roots against food-borne bacterial pathogens / Actividad Antibacteriana de la Raíz de Glycyrrhiza glabra Contra las Bacterias Patógenas Transmitidas por los Alimentos

ABSTRACT Objective: The antibacterial activity of Glycyrrhiza glabra (Liquorice) roots was evaluated against several food-borne bacterial pathogens. Methods: The in vitro anti-bacterial activity was evaluated by determining the zone diameter of inhibition (ZDI), minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) using the aqueous. Ethanolic and methanolic extracts of the roots of Glycyrrhiza glabra. Results: Therefore, significant increase in inhibitory feature was observed because of increase in extracts concentration. In addition, the aqueous extract was more effective than the others; while, among the tested bacteria, Staphylococcus aureus and Pseudomonas aeruginosa were the most sensitive and the most resistant, respectively. Conclusion: Extracts of Glycyrrhiza glabra roots can potentially be used in the pharmaceutical and food industries as preservatives or antimicrobial agents.

RESUMEN Objetivo: La actividad antibacteriana del extracto de raíces de Glycyrrhiza glabra (regaliz) fue evaluada frente a varias bacterias patógenas trasmitidas por los alimentos. Métodos: La actividad antimicrobiana se evalúa determinando el diámetro de la zona de inhibición (DZI), y la concentración bactericida mínima (CBM). Extractos acuosos, etanólicos y metanólicos de la raíz de Glycyrrhiza glabra fueron analizados en su actividad antibacteriana in vitro. Resultados: Por lo tanto, se observó un aumento significativo en la característica inhibitoria debido al aumento en la concentración de extractos. Además, el extracto acuoso fue más eficaz que los otros, en tanto que, entre las bacterias probadas, Staphylococcus aureus y Pseudomonas aeruginosa fueron las más sensibles y las más resistentes, respectivamente. Conclusión: Los resultados sugieren que los extractos de raíz de Glycyrrhiza glabra tienen un uso potencial en la industria farmacéutica y alimentaria, y pueden ser útiles como conservantes o agentes antimicrobianos.

Microbial Etiology and Antibiogram of SSIs following LSCS in a Tertiary Care Hospital in Amritsar

Surgical Site Infections (SSIs) are the second most commonly reported nosocomial infections after Urinary tract infection following a delivery by caesarean section. They add significantly to the morbidity, mortality and socioeconomic consequences in both patients as well as health care systems. Aim: To study the microbial etiology and antibiogram of post caesarean SSIs. Methods: This prospective study was conducted on pregnant women undergoing elective/emergency caesarean section irrespective of the indication during a study period from Jan 2017 to June 2018. 250 pregnant women undergoing LSCS developing signs and symptoms suggestive of SSI till the 30th post-operative day were included as cases. Under all aseptic precautions, two pus swabs were collected from every post caesarean women enrolled in the study. The first swab was used for gram staining while the other was inoculated on 5% blood agar and Macconkey agar. The isolates were identified by their colony characters and their standard biochemical tests. Antimicrobial susceptibility pattern was performed on Mueller Hinton agar plates by Kirby Bauer disc diffusion methods and interpreted as per latest CLSI guidelines. Results: Among 250 cases suspected of having SSIs, 93 aerobic isolates were obtained. Staphylococcus aureus (41.9%) was the most common organism isolated followed by Acinetobacter baumanii (15.1%), Escherichia coli(11.8%) Klebsiella pneumoniae(9.7%). 15/39(38.4%) of Staphylococcus aureus were observed to be Methicillin resistant. 15/45 (33.3%) of Gram negative isolates were Extended Spectrum Beta- Lactamase producers. Conclusion: Since MRSA is the leading cause of post caesarean SSI and the incidence of multidrug resistant pathogens as a cause of post-operative wound infections is rising with the increased prevalence of ESBL, therefore rational and judicious use of antimicrobials, active surveillance and reporting of SSI, enforcement of aseptic measures, implementation of infection control strategies and their periodic review is the need of an hour to reduce the rate of post caesarean SSIs.

Bacterial pathogens of the lower respiratory tract of calves from Brazilian rural settlement herds and their association with clinical signs of bovine respiratory disease / Patógenos bacterianos do trato respiratório inferior de bezerros criados em rebanhos de assentamentos brasileiros e sua associação com os sinais clínicos da doença respiratória dos bovinos

Bovine respiratory disease (BRD) is considered the major cause of economic losses in dairy and beef cattle production. The study aimed to detect the most important bacteria related to respiratory disease in tracheobronchial fluid samples of healthy and dairy calves with clinical signs of BRD in Brazilian rural settlements. Hundred and forty-one mongrel dairy calves were randomly selected from 42 family farm dairy herds from Brazilian settlements. Physical examination was performed and calves were classified as healthy (n=100) and BRD (n=41). Tracheobronchial fluid samples were collected. Isolation and molecular detection of Mycoplasma dispar, M. bovis and M. mycoides subsp. mycoides SC besides isolation of other aerobic bacteria were performed. Abnormal lung sounds (crackle/snoring/whistle), mucopurulent/purulent nasal discharge, body temperature >39.5°C and respiratory rate >40 breaths/min were higher in BRD calves compared to healthy calves (P<0.05). Bacillus sp., Staphylococcus intermedius and non-fermentative Gram-negative were the most prevalent bacteria isolated. Non-identified species from Enterobacteriaceae family was higher in BRD calves compared to healthy calves (P<0.05). Mollicutes were isolated in 7.4% of samples and only M. dispar was detected. Mollicutes was associated with purulent/mucopurulent nasal discharge (P=0.017). Pantoea agglomerans was associated to tachypnea (P=0.020), and Streptococcus spp. was associated with hyperthermia. Statistical tendencies were observed to M. dispar and tachypnea (P=0.066), and P. agglomerans and tachycardia (P=0.066). The obtained results describe the microorganisms found in tracheobronchial fluid of calves with BRD in some herds of Brazilian family farming and their relation to clinical signs of BRD.(AU)

A doença respiratória dos bovinos (DRB) é considerada a principal causa de perdas econômicas nas produções de leite e carne. O objetivo deste estudo foi detectar as mais importantes bactérias relacionadas a doença respiratória presentes em amostras de lavado traqueobrônquico de bezerros sadios e com sinais clínicos da DRB de assentamentos brasileiros. Cento e quarenta e um bezerros leiteiros sem raça definida foram randomicamente selecionados de 42 rebanhos leiteiros de assentamentos brasileiros. Exame físico foi realizado e os animais foram classificados em sadios (n=100) e com DRB (n=41). Amostras de lavado traqueobrônquico foram coletadas. Foram realizados o isolamento e a detecção molecular de Mycoplasma dispar, M. bovis e M. mycoides subsp. mycoides SC além de isolamento de outras bactérias aeróbias. Ruídos pulmonares anormais (crepitação/ ronco/sibilo), secreção nasal mucopurulenta/purulenta, temperatura corporal >39.5°C e frequência respiratória >40 movimentos respiratórios/min foram observados com maior frequência em bezerros com DRB comparado aos animais sadios (P<0.05). Bacillus sp, Staphylococcus intermedius e bactérias Gram-negativas não fermentadoras foram as bactérias mais prevalentes. Bactérias da família Enterobacteriaceae cuja espécie não fora identificada foram mais frequentes em bezerros com DRB comparado aos bezerros sadios (P<0.05). Mollicutes foram isolados em 7,4% das amostras e somente M. dispar foi detectado. Mollicutes foi associado à secreção nasal purulenta/mucopurulenta (P=0.017). Pantoea agglomerans foi associada a taquipneia (P=0.020), e Streptococcus spp. Foi associado a hipertermia. Tendência estatística foi observada para M. dispar e taquipneia (P=0.066), e P. agglomerans e taquicardia (P=0.066). Os resultados obtidos descrevem os micro-organismos encontrados no lavado traqueobrônquico de bezerros com DRB em rebanhos de agricultura familiar brasileira e sua relação com as manifestações clínicas da DRB.(AU)

Comparing invasive effects of five foodborne bacterial pathogens in human embryonic intestine 407 cells and human ileocecum HCT-8 cells

Objective: To refine the infectious doses of enteric bacterial pathogens in animal assays and vaccine clinical trials by studying the invasion kinetics of five bacterial pathogens with human intestinal cells. Methods: Utilizing in vitro cultured cell invasion assays with gentamicin-killing step, the invasive effects were analyzed in foodborne pathogens including Salmonella, Shigella, Yersinia, Escherichia coli (E. coli) O157 and opportunistic pathogens Cit-robacter in human embryonic intestine 407 cells and ileocecum HCT-8 cells at multi-plicities of infection(MOIs)of 0.04–4 000.00 E.coli HS served as a noninvasive control. Results: The study results showed that the bacterial invasive efficiency and the average number of internalized bacteria per host cell changed with different starting MOIs.Higher starting MOIs did not always produce more bacterial internalization. The bacterial in-vasion effects varied with different bacterial strains and host cell lines.E.coli O157:H7 did invade human ileocecum HCT-8 cells. Conclusions: This study shows that these bacteria possess different invasive patterns at various starting MOIs and also in different cell lines.The results could help to figure out the appropriate infectious doses of the bacteria in animal assays and in vaccine clinical trials.The bacterial invasion kinetics is also valuable in evaluating the safety and efficacy of live attenuated bacterial vaccines.

Bacteriological profile of surgical site infections

Background: The susceptibility and severity of Rheumatoid arthritis are determined by both genetic and environmental factors. Dermatoglyphic patterns of individuals which are formed early in the fetal life are also determined by both genetic and environmental factors. Since both are genetically acquired and environmentally modified, it has been shown that there are particular dermatoglyphic patterns associated with Rheumatoid arthritis. If it is so, dermatoglyphics can serve as an additional tool in the early diagnosis and management of such a disabling disease like rheumatoid arthritis. Although there are some studies which mentioned dermatoglyphic pattern variation in the disease, the results are contradicting. Therefore, the present study was undertaken to find out a possible correlation of some quantitative and qualitative dermatoglyphic variables with Rheumatoid Arthritis. Aim of the study: To study the dermatoglyphic patterns in patients with Rheumatoid Arthritis and control population and to study the correlation between dermatoglyphic patterns and Rheumatoid Arthritis. Materials and methods: We studied 60 patients with Rheumatoid arthritis and 60 controls. All were subjected to detailed medical history and clinical examination. Both quantitative (finger ridge count and pattern intensity) and qualitative (fingerprint pattern) dermatoglyphic parameters were studied and the same were compared with age, sex and disease matched controls. Results: Out of the total 60 cases 12(20%) were male and 48(80%) were females. Of the total 60 controls, 12(20%) were males and 48(80%) were females. Analysis of the qualitative parameters revealed: Significant increase in the number of whorls in both the hands of female patients cases compared to the controls (p-value for right hand 0.001, p-value for left hand 0.004). The decrease in the number of radial loops in both the hands of male and female patients and the decrease was more in the left hand in males and right hand in females (p-value male left hand 0.002, female right hand Saritha K. Narayanan, Christopher C. Pais, Pradeep Kumar Shenoy. Use of palmar dermatoglyphics in rheumatoid arthritis - A case-control study. IAIM, 2017; 4(12): 70-76. Page 71 0.003). Decrease in the number of arches in the left hand of female patients compared to the controls (p-0.10). Analysis of the quantitative parameters showed: A statistically significant increase in the finger ridge count of individual hand and the total finger ridge count in both male and female patients compared to the controls (p-value males: right hand 0.003, left hand 0.004, right plus left hand 0.002; p-value females right hand.0000, left hand 0.000, right plus left hand 0.000). A statistically significant increase in the pattern intensity of fingers in female patients compared to the controls (p-value: right hand 0.006, left hand 0.001, right plus left hand 0.000). Conclusion: The findings of this work demonstrate the association between some of the qualitative and quantitative parameters of dermatoglyphics and Rheumatoid arthritis suggesting that dermatoglyphics can represent an anatomical, non-invasive, inexpensive tool for screening high-risk population and thus facilitate early detection and management.

Assessment and incidence of fish associated bacterial pathogens at hatcheries of Azad Kashmir, Pakistan

Abstract Fish is the most indispensable source of proteins for individuals and have high nutritional value. On the other hand, the fish culturing raised issues of fish health due to close contact between the aquatic environment and the fish pathogens. So, the aim of the current study was to identify the bacterial pathogens and screen the injured Rainbow trout rearing in different trout hatcheries run under fisheries department of the government of Azad Jammu and Kashmir, Pakistan. Seven bacterial pathogens such as Shigella flexneri, Enterobacter amnigenus, Salmonella Typhimurium, Serratia odorifera, Pseudomonas aeruginosa, Streptococcus pyogenes, and Bacillus cereus were isolated and identified. Results revealed that the injury of fish specimens was due to overcrowding. Instead of rainbow coloration, specimens have darker black in color. The water of ponds was not clean and clear and such conditions was because of the greater quantity of feed thrown in the water. It was concluded that poor hygienic water condition and overloading allowed the opportunistic bacterial contaminations to succeed which cause a serious threat to hatcheries.

Resumo O peixe é a fonte mais indispensável de proteínas para os indivíduos e tem alto valor nutricional. Por outro lado, a cultura dos peixes levantou questões sobre a saúde dos peixes devido ao próximo contato entre o ambiente aquático e os agentes patogênicos desses peixes. Assim, o objetivo do presente estudo foi identificar os patógenos bacterianos e rastrear a criação da truta arco-íris que apresentou lesões em diferentes incubadoras de trutas, com supervisão do departamento de pesca do governo de Azad Jammu e Caxemira, Paquistão. Sete patógenos bacterianos foram isolados e identificados, tais como: Shigella flexneri, Enterobacter amnigenus, Salmonella typhimurium, Serratia odorifera, Pseudomonas aeruginosa, Streptococcus pyogenes e Bacillus cereus. Os resultados revelaram que a lesão de espécimes de peixes foi devido à superlotação. Em vez da coloração do arco-íris, os espécimes tiveram uma coloração preta mais escura. A água das lagoas não era limpa e nem clara, e tais condições ocorreram devido a maior quantidade de alimento lançada na água. Concluiu-se que a precária condição higiênica da água e também a sobrecarga permitiram que as contaminações bacterianas oportunistas fossem bem-sucedidas, causando séria ameaça às incubadoras.

Comparing invasive effects of five foodborne bacterial pathogens in human embryonic intestine 407 cells and human ileocecum HCT-8 cells

Objective To refine the infectious doses of enteric bacterial pathogens in animal assays and vaccine clinical trials by studying the invasion kinetics of five bacterial pathogens with human intestinal cells. Methods Utilizing in vitro cultured cell invasion assays with gentamicin-killing step, the invasive effects were analyzed in foodborne pathogens including Salmonella, Shigella, Yersinia, Escherichia coli (E. coli) O157 and opportunistic pathogens Citrobacter in human embryonic intestine 407 cells and ileocecum HCT-8 cells at multiplicities of infection (MOIs) of 0.04–4 000.00 E. coli HS served as a noninvasive control. Results The study results showed that the bacterial invasive efficiency and the average number of internalized bacteria per host cell changed with different starting MOIs. Higher starting MOIs did not always produce more bacterial internalization. The bacterial invasion effects varied with different bacterial strains and host cell lines. E. coli O157:H7 did invade human ileocecum HCT-8 cells. Conclusions This study shows that these bacteria possess different invasive patterns at various starting MOIs and also in different cell lines. The results could help to figure out the appropriate infectious doses of the bacteria in animal assays and in vaccine clinical trials. The bacterial invasion kinetics is also valuable in evaluating the safety and efficacy of live attenuated bacterial vaccines.

Terapia de manutenção periodontal: perfil microbiológico e efeito do tabagismo na perda dental / Periodontal maintenance therapy: microbial profile and effects of smoking on tooth loss

Esta Tese reporta dois estudos distintos relacionados à terapia de manutenção periodontal (TMP): achados microbiológicos longitudinais de 6 anos relacionados à cooperação dos indivíduos e efeito do tabagismo na perda dental. A justificativa para estes estudos é baseada em dois aspectos pontuais: (1) Embora a associação do tabagismo como fator de risco para a periodontite e pior resposta à TMP tenha sido demonstrada em muitos estudos, o efeito isolado deste sobre a perda dental em indivíduos submetidos a TMP ainda não foi reportado em revisões sistemáticas; (2) Poucos estudos têm sido conduzidos avaliando mudanças longitudinais na microbiota subgengival de indivíduos em TMP e, até o momento, nenhum estudo relacionou níveis de bactérias associadas à periodontite com o grau de cooperação dos indivíduos em TMP. Neste sentido, os objetivos deste estudo são apresentados em dois artigos científicos: (1) realizar uma revisão sistemática e meta-análise do efeito do tabagismo na perda dental em indivíduos em TMP, com a seguinte questão focal: ¿Qual é o efeito do tabagismo na perda dental em indivíduos em terapia de manutenção periodontal?¿; (2) avaliar longitudinalmente, durante 6 anos, o efeito da cooperação em TMP na frequência das bactérias Actinomyces naeslundii, Porphyromonas gingivalis, Tanerella forsythia e Treponema dentícola. Na revisão sistemática as bases de dados MEDLINE, WEB OF SCIENCE, COCHRANE LIBRARY e SCOPUS foram pesquisadas, incluindo artigos até fevereiro de 2017. A estratégia de busca identificou 728 referências. Após a remoção das duplicatas, restaram 591 para a seleção baseada nos títulos e resumos, e desses, foram selecionados 36 para leitura completa do texto. Após a leitura, foram incluídos 10 artigos para revisão sistemática e 3 artigos para meta-análise. A qualidade da evidência científica foi moderada para indivíduos tabagistas em TMP apresentarem maior chance de perda dental que indivíduos não tabagistas. A metodologia do estudo microbiológico englobou um total de 56 indivíduos, todos recrutados de um coorte prospectivo com 212 participantes de um programa de TMP. Estes indivíduos foram acompanhados durante 6 anos, em 5 tempos de avaliações: T1 (anterior à terapia periodontal ativa), T2 (após a terapia periodontal ativa), T3 (última visita à TMP em 2 anos), T4 (última visita à TMP em 4 anos) e T5 (última visita à TMP em 6 anos). Assim, 28 indivíduos cooperadores regulares (CR) foram randomizados e pareados, pelo gênero e idade, a 28 indivíduos cooperadores irregulares (CI). Durante a TMP, os exames periodontais avaliaram os parâmetros clínicos: índice de placa (IP), profundidade de sondagem (PS), nível clínico de inserção (NCI) e sangramento à sondagem (SS). A quantificação de bactérias em carga total e dos níveis de A. naeslundii, P. gingivalis, T. forsythia e T. dentícola foi executada pela reação em cadeia da polimerase quantitativa (qPCR). Este estudo concluiu que CI apresentaram piores parâmetros clínicos periodontais e maiores níveis de carga bacteriana total que CR, refletindo o papel benéfico da cooperação na TMP em manter a estabilidade da condição periodontal

This Thesis report two distinct issues related to periodontal maintenance therapy (PMT): microbiological findings of 6 years related to compliance of individuas and effect of smoking on tooth loss. The rattionale for these studies is based on two specific aspects: (1) Although the association of smoking as a risk factor for periodontitis and worse response to PMT has been demonstrated in many studies, the isolated effect of smoking on tooth loss in patients undergiong PMT has not been reported in systematic reviews; (2) Few studies have been conducted on longitudinal changes in the subgingival microbiota of individuals in PMT and to date, no study related levels of bacteria to the degree of cooperation of individuls in PMT...

Screening and antibacterial efficacy of selected Indian medicinal plants

Objective: To evaluate the antibacterial efficacy of five Indian medicinal plants such as Acalypha indica L. (A. indica), Aerva lanata (L.) Juss. ex Schult. (A. lanata), Clerodendrum inerme (L.) Gaertn., Pergularia daemia (Forsk.) Chiov. and Solanum surattense Burm. f. against opportunistic bacterial pathogens isolated from HIV infected patients for the potential phytoconstituents in plant extracts. Methods: The opportunistic bacterial pathogens such as Escherichia coli (E. coli), Pseudomonas aeruginosa, Salmonella typhi and Serratia marcescens from Gram-negative group and Staphylococcus aureus from Gram-positive group were isolated from HIV infected patients. The antibacterial efficacy of ethanolic extracts of selected medicinal plants was carried out by disc diffusion method. The potential phytoconstituents of medicinal plant extracts were identified by gas chromatography and mass spectrometry (GC-MS) analysis. Results: Among the five medicinal plants tested, A. indica and A. lanata showed the significant antibacterial activity. A. indica showed potential activity against Staphylococcus aureus and E. coli. A. lanata significantly exhibited antibacterial activity against E. coli, Salmonella typhi and Pseudomonas aeruginosa. A total of 19 phytoconstituents were identified in the ethanolic extract of A. indica and A. lanata by GC-MS analysis respectively. Conclusions: The results of the present investigation revealed that A. indica and A. lanata, possessed significant antibacterial activity when compared with the other plant extracts tested. The presence of 3-O-methyl- d-glucose by GC-MS analysis in both A. indica and A. lanata extracts has not been reported elsewhere in the literature and the findings in this study could be the first one to report.

Liquid based formulations of bacteriophages for the management of waterborne bacterial pathogens in water microcosms.

Water resources are contaminated by life-threatening multidrug resistant pathogenic bacteria. Unfortunately, these pathogenic bacteria do not respond to the traditional water purification methods. Therefore, there is a need of environmentally friendly strategies to overcome the problems associated with the antimicrobial resistant bacterial pathogens. In the present study, highly potent lytic phages against multidrug-resistant Salmonella enterica serovar Paratyphi B, Pseudomonas aeruginosa and Klebsiella pneumoniae were isolated from the Pavana river water. They belonged to the Podoviridae and Siphoviridae families. These phages were purified and enriched in the laboratory. Monovalent formulations of φSPB, BVPaP-3 and KPP phages were prepared in three different liquids viz., phage broth, saline and distilled water. The phages were stable for almost 8-10 months in the phage broth at 4 °C. The stability of the phages in saline and distilled water was 5-6 months at 4 °C. All of the phages were stable only for 4-6 months in the phage broth at 30 °C. The monovalent phage formulation of φSPB was applied at MOI < 1, as disinfectant against an exponential and stationary phase cells of Salmonella enterica serovar Paratyphi B in various water microcosms. The results indicated that there was almost 80 % reduction in the log phase cells of Salmonella serovar Paratyphi B in 24 h. In stationary phase cells, the reduction was comparatively less within same period. At the same time, there was concomitant increase in the phage population by 80% in all the microcosms indicating that φSPB phage is highly potent in killing pathogen in water. Results strongly support that the formulation of φSPB in the phage broth in monovalent form could be used as an effective biological disinfectant for preventing transmission of water- borne bacterial pathogens, including antimicrobial resistant ones.

Fecal indicators and bacterial pathogens in bottled water from Dhaka, Bangladesh

Forty-six bottled water samples representing 16 brands from Dhaka, Bangladesh were tested for the numbers of total coliforms, fecal indicator bacteria (i.e., thermotolerant Escherichia coli and Enterococcus spp.) and potential bacterial pathogens (i.e., Aeromonas hydrophil, Pseudomonas aeruginos, Salmonella spp., and Shigella spp.). Among the 16 brands tested, 14 (86%), ten (63%) and seven (44%) were positive for total coliforms, E. coil and Enterococcus spp., respectively. Additionally, a further nine (56%), eight (50%), six (37%), and four (25%) brands were PCR positive for A. hydrophila lip, P. aeruginosa ETA, Salmonella spp. invA, and Shigella spp. ipaH genes, respectively. The numbers of bacterial pathogens in bottled water samples ranged from 28 ± 12 to 600 ± 45 (A. hydrophila lip gene), 180 ± 40 to 900 ± 200 (Salmonella spp. invA gene), 180 ± 40 to 1,300 ± 400 (P. aeruginosa ETA gene) genomic units per L of water. Shigella spp. ipaH gene was not quantifiable. Discrepancies were observed in terms of the occurrence of fecal indicators and bacterial pathogens. No correlations were observed between fecal indicators numbers and presence/absence of A. hydrophila lip (p = 0.245), Salmonella spp. invA (p = 0.433), Shigella spp. ipaH gene (p = 0.078), and P. aeruginosa ETA (p = 0.059) genes. Our results suggest that microbiological quality of bottled waters sold in Dhaka, Bangladesh is highly variable. To protect public health, stringent quality control is recommended for the bottled water industry in Bangladesh.

Studies on extended beta lactamase producing, biofilm forming clinical bacterial pathogens and its invitro inhibition by Actinobacterial extracts.

At present scenario, the extended spectrum beta lactamase (ESBL) producing bacterial pathogen causes various life threatening infections especially by the members of the family Enterobacteriaceae in hospital settings. In order to study the prevalence of ESBLs in Kanchipuram hospital, the bacterial strains were isolated from patients having Urinary tract infections (UTI), diabetic foot ulcer, pregnancy women’s, surgical wound infections, deep wounds, and genitourinary tract problems. Totally 40 bacterial isolates were recovered from 30 samples and the isolates were identified as Escherichia coli (45%), Pseudomonas sp, (25%) and Klebsiella sp (30%). The ESBL production was confirmed with third generation cephalosporins (cetixime, cephoxitin, ceftazidime, cetepime, ceftriaxone, ceftazidime/clavulanic acid) using the Kirby- bauer disc diffusion method and also by double disc diffusion method. The highest ESBL production was found among E. coli (42%), followed by Pseudomonas sp. (25%) and Klebsiella sp (20%). All the ESBL producer were tested for biofilm formation by tube method in which E.coli (43%) was found to be the good biofilm producer followed by the Klebsiella sp (31%) and Pseudomonas sp (25%). An attempt was also made to study the in-vitro inhibition of biofilm forming ESBL pathogens by actinobacterial extracts by disc diffusion method. Of the five actinobacterial extracts tested, extracts produced from the strain MA7 inhibited (8-12 mm zone of inhibition) all the biofilm forming ESBL pathogens. Further purification and characterisation of active compound from actinobacterial strain MA7 is in progress.