Detection of oral spirochete (Treponema denticola) by modified Fontana staining technique from sub-gingival plaques of patients suffering from periodontal disease and to analyse the morphology of spirochetes with respect to the disease’s status

Background:A large proportion of fixed dose combinations (FDCs) are manufactured and used widely in Nepal. This study aimed to evaluate the FDCs and its utilization in medicine department of tertiary care hospital. Methods:A cross-sectional study was conducted for 50 days among admitted patients in the medicine department of tertiary care hospital, Nepal. A predesigned form was used to collect the data at the time of patient discharge. Only the oral FDCs were selected for study.Microsoft Excel 2007 was used for statistical analysis and data were presented as number and percentage in tabulated and figure forms.Results:Oral FDCs were used in 27.08% of admitted patients. A total of 295 FDCs were prescribed in 208 patients with 44 FDC items in 58 different brand names. Categorically, the most commonly used FDCs were of analgesics (34.24%) followed by antibiotics (25.76%) and vitamin supplements (22.71%). The 27.27% of FDCs prescribed contain more than two active pharmaceutical ingredients (APIs) up to nine and the highest number of APIs were found in vitamin supplements. All FDCs were prescribed in the brand names. The very few 2.27% and 4.55% of FDCs were prescribed from the essential medicine list of Nepal and world health organization, respectively.Conclusions:The use of FDCs listed in essential medicine list was very poor. Similarly, generic prescribing was also zero. The regulatory body must study the rationality of FDC before production, marketing, importing, and utilization in hospital.

Leptospira patógena en murciélagos de Campeche y Yucatán, México / Pathogenic Leptospira in bats from Campeche and Yucatán, Mexico

RESUMEN Objetivo. Reportar la infección con Leptospira en ríñones de murciélagos de Campeche y Yucatán, México, a través de la amplificación por PCR de dos fragmentos distintos del gen 16S RNA ribosomal. Materiales y métodos. Se realizaron capturas en un sitio de Campeche y dos de Yucatán. A los murciélagos capturados se les aplicó la eutanasia y se les realizó una necropsia para recolectar tejido renal que se usó en la extracción de ADN total. Se realizaron dos PCR convencionales para la amplificación de los fragmentos de 16S RNA ribosomal. Se obtuvieron las secuencias de algunos productos positivos y se analizaron con herramientas bioinformáticas para identificar la especie infectante de Leptospira. Resultados. Se capturaron 69 murciélagos pertenecientes a cuatro familias y a ocho especies distintas. La familia con mayor diversidad fue Phyllostomidae con cinco especies. La especie con mayor frecuencia de captura fue Artibeusjamaicensis (41, 59.4%). Las PCR arrojaron una frecuencia global de infección de 21.7%. Las especies infectadas fueron A. jamaicensis, Pteronotus parnellii y Chiroderma villosum. El análisis bioinformático arrojó un 99.0% de identidad para Leptospira noguchii, Leptospira borgpetersenii y Leptospira santarosai. Conclusiones. Algunas especies de murciélagos de Yucatán y Campeche son portadores renales de leptospiras patógenas, por lo que podrían participar en el ciclo silvestre de transmisión en la región. La frecuencia de infección encontrada en los riñones de los murciélagos utilizados es mayor en comparación con aquellas obtenidas en otros reservorios de Yucatán y Campeche. Nuevas especies de murciélagos son reportadas como portadores de Leptospira para México.

ABSTRACT Objective. To report the infection with Leptospira in the kidneys of bats from Campeche and Yucatán, Mexico, through the amplification by PCR of two different 16S RNA ribosomal gene fragments. Materials and methods. Bat captures were carried out at one site in Campeche and two sites in Yucatán. Euthanasia was applied to the captured bats and a necropsy was performed to collect a renal tissue sample that was used in the total DNA extraction. Two different conventional PCR were performed for the amplification of the 16S RNA ribosomal gene fragments. Some sequences from positive products were obtained and analyzed with bioinformatics tools to identify the infectious species of Leptospira. Results. Sixty-nine bats belonging to four families and eight different species were captured. The family with the greatest diversity was Phyllostomidae with five species. The most captured species was Artibeus jamaicensis (41, 59.4%). Both PCR showed a global infection frequency of 21.7%. The infected species were A. jamaicensis, Pteronotus parnellii, and Chiroderma villosum. The bioinformatic analysis of the positive products yielded a 99.0% identity for Leptospira noguchii, Leptospira borgpetersenii, and Leptospira santarosai. Conclusions. Some bat species of Yucatán and Campeche, Mexico, are renal carriers of pathogenic Leptospira, therefore participating in the transmission cycle in the region. The frequency of infection found in the renal tissue of the captured bats is higher than the one obtained from other reservoirs captured in Yucatán and Campeche. New species of bats are reported as renal Leptospira carriers in Mexico.

Evaluation of fecal microbiomes associated with obesity in captive cynomolgus monkeys (Macaca fascicularis)

Microorganisms play important roles in obesity; however, the role of the gut microbiomes in obesity is controversial because of the inconsistent findings. This study investigated the gut microbiome communities in obese and lean groups of captive healthy cynomolgus monkeys reared under strict identical environmental conditions, including their diet. No significant differences in the relative abundance of Firmicutes, Bacteroidetes and Prevotella were observed between the obese and lean groups, but a significant difference in Spirochetes (p < 0.05) was noted. Microbial diversity and richness were similar, but highly variable results in microbial composition, diversity, and richness were observed in individuals, irrespective of their state of obesity. Distinct clustering between the groups was not observed by principal coordinate analysis using an unweighted pair group method. Higher sharedness values (95.81% ± 2.28% at the genus level, and 79.54% ± 5.88% at the species level) were identified among individual monkeys. This paper reports the association between the gut microbiome and obesity in captive non-human primate models reared under controlled environments. The relative proportion of Firmicutes and Bacteroidetes as well as the microbial diversity known to affect obesity were similar in the obese and lean groups of monkeys reared under identical conditions. Therefore, obesity-associated microbial changes reported previously appear to be associated directly with environmental factors, particularly diet, rather than obesity.

Comparison of Immunoprotection of Leptospira Recombinant Proteins with conventional vaccine in experimental animals.

Leptospirosis is a bacterial disease caused by bacteria of the genus Leptospira affecting humans and animals. Untreated leptospirosis may result in severe kidney damage, meningitis, liver failure, respiratory distress, and even death. Virulent leptospirosis can rapidly enter kidney fibroblasts and induce a programmed cell death. Thus, it is a challenge for immunologists to develop an effective and safe leptospirosis vaccine.Here, we compared the commercial canine leptospira vaccine and recombinant proteins (OmpL1 and LipL41) with and without adjuvant in terms of immune response and challenge studies in hamsters and immune response studies alone in experimental dogs. The outer membrane proteins viz., lipL41 and OmpL1 of leptospira interrogans serovars icterohaemorrhagiae were amplified. The primers were designed in such a way that amplified products of OmpL1 and lipL41 were ligated and cloned simultaneously into a single vector. The cloned products were expressed in E.coli BL21 cells. The immunoprotection studies were conducted for both recombinant proteins and commercial vaccine. The challenge experiment studies revealed that combination of both rLip41 and rOmpL1 and commercial vaccine gave 83% and 87% protection, respectively. Histopathological investigation revealed mild sub lethal changes were noticed in liver and kidney in commercially vaccinated group alone. The immune responses against recombinant leptospiral proteins were also demonstrated in dogs.

A case report of Ludwig’s angina.

Ludwig angina is a rapidly progressing submaxillary, submandibular, and sublingual necrotizing cellulitis of the floor of the mouth that can have lethal consequences due to airway obstruction Various aerobic and anaerobic microorganisms, and less often fungi, have been implicated to cause Ludwig angina, including oral flora such as Streptococci and Staphylococci. Early recognition and use of parenteral antibiotics can prevent mortality and morbidity. We report a case of 30 years old male who was admitted to hospital with chief complaints of neck swelling, toothache, dysphagia and difficulty in opening mouth. Blood counts showed leukocytosis with neutrophilia along with raised ESR. Pus was drained after incision in submental and submandibular space and was transported to Microbiology department for further processing. Gram staining of pus showed many pus cells, spirochetes and fusiform shaped bacilli.

Digital dermatitis of the accessory digits of dairy cows / Dermatite digital nos dígitos acessórios de vacas leiteiras

This report characterizes the digital dermatitis (DD) lesions in the accessory digits of dairy cows and presents data on the applied therapy. Fifteen Holstein cattle with DD affecting the accessory digits of the hindlimbs from four dairy farms with previous history of DD were evaluated. Lesions were excised, the wounds were sutured, and a topical application of oxytetracycline powder covered by bandaging was associated with a single parenteral administration of long acting oxytetracycline IM (20mg/kg). Tissue samples were obtained for histopathology and transmission electronic microscopy (TEM). Lesions from all the animals were recuperated 15 days after surgical procedure. Overal, most DD lesions were papillomatous epidermal projections or wartlike verrucous lesions. Histopathologically, samples revealed hyperplasia of epidermis with hyperkeratosis, several mitoses in the stratum basale and elongated rete ridges in the superficial and middle dermis. TEM revealed long, thin spirochete-like bacteria. Morphologic features of lesions and its response to therapy were comparable to those described for DD.

Esse estudo caracteriza lesões de dermatite digital (DD) nos dígitos acessórios de vacas leiteiras, além de apresentar a terapia aplicada. Foram utilizados 15 bovinos leiteiros da raça Holandês com DD nos dígitos acessórios dos membros pélvicos. Os animais eram provenientes de quatro fazendas leiteiras com histórico prévio de dermatite digital (DD). Todos os quinze animais foram tratados da mesma forma: após excisão das lesões e sutura das feridas cutâneas, aplicou-se oxitetraciclina pó topicamente sob bandagem e oxitetraciclina (20mg/kg) de longa ação, via intramuscular. Obtiveram-se amostras de tecidos para histopatologia, inclusive por microscopia eletrônica de transmissão (MET). Observou-se cicatrização em todos os animais após 15 dias do procedimento cirúrgico. A maioria das lesões macroscópicas foram projeções papilomatosas ou em forma de verrugas. Os achados histopatológicos de todas as amostras revelaram hiperplasia da epiderme com hiperceratose, inúmeras mitoses no estrato basal, com invasões alongadas em forma de rede na derme superficial e intermediária. A MET evidenciou organismos com formas longas, afiladas e espiraladas, presumivelmente espiroquetas. Tanto as características morfológicas, quanto a resposta à terapia das lesões foram comparáveis às descritas para DD.

How to Denominate "Four Pathogens and One Bacterium" / 微生物学通报

Mycoplasma, Chlamydia, Rickettsia, Spirochetes and Actinobacteria were generally called "four pathogens and one bacterium". It was always difficult to be denominated and classified rightly in textbooks, while it was also a key interfering with students to grasp the concept of bacteria exactly. So we raise the question and hope to learn from each other by an exchange of views here.

Borrelia theileri: observação em carrapatos do gênero Boophilus microplus no município de Guaíba, RS, Brasil / Borrelia theileri: observation on Boophilus microplus ticks in Guaiba, RS, Brazil

Espiroquetas da espécie Borrelia theileri identificadas em uma estirpe de carrapatos Boophilus microplus provenientes do município de Guaíba, RS. A observação ocorreu no exame de hemolinfa de fêmeas adultas com 10 dias pós-repleçâo, corada por Giemsa. Não foram observadas espiroquetas em ovos provenientes de teleóginas infectadas. A detecção da estirpe infectada sugere a presença de borreliose em rebanhos bovinos, fato que eventualmente pode interferir em resultados de diagnóstico ou tornar-se motivo de preocupação em produtos derivados de sangue bovino tais como vacinas vivas contra anaplasmose e babesiose bovina.

Spirochetes of species Borrelia theileri were identifica in afield-strain of the caule tick Boophilus microplus, in Guaíba, RS, Brazil. Hemolymph smears from females 10 days post-repletion were collected by gentty section of the tarsal-metatarsaijoint, and dropped onto a microscope slide, and stained by Giemsa. No spirochetes were observed in eggs squashed and stained by Giemsa from the same infected strain. The detection of B. microplus adult females infected with Borrelia theileri suggesfs the likely presence of borreliosis in bovine heras what might eventually interfere with the interpretation of diagnosis results or become cause for concern in blood products such as anaplasmosis and babesiosis live vaccines.