Percentage Incidences of Bacteria in Mahseer (Tor putitora), Silver carp (Hypophthalmichthys molitrix) Fish Collected from Hatcheries and Local Markets of District Malakand and Peshawar of Khyber Pakhtunkhwa, Pakistan

Abstract Fish is the main source of animal protein for human diet. The aim of this study was to find out prevalence of pathogenic bacteria of two selected economically important fish of Pakistan namely Mahseer (Tor putitora) and Silver carp (Hypophthalmichthys molitrix). Live fish samples from hatcheries and dead fish samples from different markets of study area were randomly collected. The fish samples were analyzed for isolation, identification and prevalence of bacteria. The isolated bacteria from study fish were identified through biochemical test and about 10 species of pathogenic bacteria were identified including the pathogenic bacteria to human and fish namely, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. and Salmonella spp. The bacterial percentage frequency of occurrence in Silver carp and Mahseer fish showed Pseudomonas aeruginosa 21.42%, Staphylococcus epidermidis 17.85%, Escherichia coli 11.90%, Staphylococcus aureus 9.52%, Citrobacter spp. 9.52%, Serratia spp. 8.33%, Streptococcus iniae 7.14%, Stenotrophomonas spp. 5.95%, Bacillus spp. 4.76% and Salmonella spp. 3.57%. The study revealed that Fish samples of Mahseer and Silver carp that were collected from markets have found more isolates (10 bacterial species) than did the fresh fish pond samples (03 bacterial species) of hatcheries. The occurrence of pathogenic bacteria in study fish showed risk factor for public health consumers.

Resumo O peixe é a principal fonte de proteína animal para a alimentação humana. O objetivo deste estudo foi descobrir a prevalência de bactérias patogênicas de dois peixes economicamente importantes selecionados do Paquistão, nomeadamente Mahseer (Tor putitora) e carpa prateada (Hypophthalmichthys molitrix). Amostras de peixes vivos de incubatórios e amostras de peixes mortos de diferentes mercados da área de estudo foram coletadas aleatoriamente. As amostras de peixes foram analisadas quanto ao isolamento, identificação e prevalência de bactérias. As bactérias isoladas dos peixes do estudo foram identificadas através de testes bioquímicos e cerca de 10 espécies de bactérias patogênicas foram identificadas incluindo as bactérias patogênicas para humanos e peixes, nomeadamente, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. e Salmonella spp. A porcentagem de freqüência de ocorrência bacteriana em carpa prateada e peixes Mahseer mostrou Pseudomonas aeruginosa 21,42%, Staphylococcus epidermidis 17,85%, Escherichia coli 11,90%, Staphylococcus aureus 9,52%, Citrobacter spp. 9,52%, Serratia spp. 8,33%, Streptococcus iniae 7,14%, Stenotrophomonas spp. 5,95%, Bacillus spp. 4,76% e Salmonella spp. 3,57%. O estudo revelou que as amostras de peixes de Mahseer e carpa prateada coletadas nos mercados encontraram mais isolados (10 espécies bacterianas) do que as amostras de peixes frescos (03 espécies bacterianas) de incubatórios. A ocorrência de bactérias patogênicas nos peixes do estudo apresentou fator de risco para consumidores de saúde pública.

Percentage Incidences of Bacteria in Mahseer (Tor putitora), Silver carp (Hypophthalmichthys molitrix) Fish Collected from Hatcheries and Local Markets of District Malakand and Peshawar of Khyber Pakhtunkhwa, Pakistan / Incidências percentuais de bactérias em Mahseer (Tor putitora), carpa-prateada (Hypophthalmichthys molitrix), peixes coletados em incubatórios e mercados locais do distrito de Malakand e Peshawar de Khyber Pakhtunkhwa, Paquistão

Abstract Fish is the main source of animal protein for human diet. The aim of this study was to find out prevalence of pathogenic bacteria of two selected economically important fish of Pakistan namely Mahseer (Tor putitora) and Silver carp (Hypophthalmichthys molitrix). Live fish samples from hatcheries and dead fish samples from different markets of study area were randomly collected. The fish samples were analyzed for isolation, identification and prevalence of bacteria. The isolated bacteria from study fish were identified through biochemical test and about 10 species of pathogenic bacteria were identified including the pathogenic bacteria to human and fish namely, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. and Salmonella spp. The bacterial percentage frequency of occurrence in Silver carp and Mahseer fish showed Pseudomonas aeruginosa 21.42%, Staphylococcus epidermidis 17.85%, Escherichia coli 11.90%, Staphylococcus aureus 9.52%, Citrobacter spp. 9.52%, Serratia spp. 8.33%, Streptococcus iniae 7.14%, Stenotrophomonas spp. 5.95%, Bacillus spp. 4.76% and Salmonella spp. 3.57%. The study revealed that Fish samples of Mahseer and Silver carp that were collected from markets have found more isolates (10 bacterial species) than did the fresh fish pond samples (03 bacterial species) of hatcheries. The occurrence of pathogenic bacteria in study fish showed risk factor for public health consumers.

Resumo O peixe é a principal fonte de proteína animal para a alimentação humana. O objetivo deste estudo foi descobrir a prevalência de bactérias patogênicas de dois peixes economicamente importantes selecionados do Paquistão, nomeadamente Mahseer (Tor putitora) e carpa prateada (Hypophthalmichthys molitrix). Amostras de peixes vivos de incubatórios e amostras de peixes mortos de diferentes mercados da área de estudo foram coletadas aleatoriamente. As amostras de peixes foram analisadas quanto ao isolamento, identificação e prevalência de bactérias. As bactérias isoladas dos peixes do estudo foram identificadas através de testes bioquímicos e cerca de 10 espécies de bactérias patogênicas foram identificadas incluindo as bactérias patogênicas para humanos e peixes, nomeadamente, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. e Salmonella spp. A porcentagem de freqüência de ocorrência bacteriana em carpa prateada e peixes Mahseer mostrou Pseudomonas aeruginosa 21,42%, Staphylococcus epidermidis 17,85%, Escherichia coli 11,90%, Staphylococcus aureus 9,52%, Citrobacter spp. 9,52%, Serratia spp. 8,33%, Streptococcus iniae 7,14%, Stenotrophomonas spp. 5,95%, Bacillus spp. 4,76% e Salmonella spp. 3,57%. O estudo revelou que as amostras de peixes de Mahseer e carpa prateada coletadas nos mercados encontraram mais isolados (10 espécies bacterianas) do que as amostras de peixes frescos (03 espécies bacterianas) de incubatórios. A ocorrência de bactérias patogênicas nos peixes do estudo apresentou fator de risco para consumidores de saúde pública.

Isolation and characterization of bacteria residing in the oral, gut, and fecal samples of different pheasant species / Isolamento e caracterização de bactérias residentes nas amostras orais, intestinais e fecais de diferentes espécies de faisão

There is a paucity of research conducted on microbial prevalence in pheasants. The microbiota of captive birds has zoonotic significance and must be characterize. Present study is therefore planned to assess the microbiota from oral, fecal and gut content of captive avian species. It will be helpful in characterization of harmful microbes. Different samples taken from oral, gut and feces of ring-necked pheasants (Phasianus colchicus), green pheasants (Phasianus versicolor), golden pheasant (Chrysolophus pictus) and silver pheasant (Lophura nycthemera). Samples were collected, diluted, and inoculated onto different agar plates (MacConkey, SS agar, MSA and nutrient agar) for cultivation of bacterial species. Colonies of E.coli, Staphylococcus spp. Brachyspira spp. and Campylobacter spp were observed based on colony morphology. Colony forming unit showed E. coli as frequently found bacteria in fecal, oral and gut contents of all the above pheasants. The overall significance difference was found among bacterial species of golden pheasants, green pheasant, ring-necked pheasant, and silver pheasants. It was concluded that E.coli is predominant isolated from heathy pheasants followed by Campylobacter, Staphylococcus and Brachyspira.

Há uma escassez de pesquisas realizadas sobre a prevalência microbiana em faisões. A microbiota de aves em cativeiro tem significado zoonótico e deve ser caracterizada. O presente estudo está, portanto, planejado para avaliar a microbiota do conteúdo oral, fecal e intestinal de espécies aviárias em cativeiro. Será útil na caracterização de micróbios nocivos. Diferentes amostras retiradas da boca, intestino e fezes de faisões de pescoço redondo (Phasianus colchicus), faisões verdes (Phasianus versicolor), faisões dourados (Chrysolophus pictus) e faisão prateado (Lophura nycthemera). As amostras foram coletadas, diluídas e inoculadas em diferentes placas de ágar (MacConkey, ágar SS, MSA e ágar nutriente) para o cultivo de espécies bacterianas. Colônias de E. coli, Staphylococcus spp., Brachyspira spp. e Campylobacter spp foram observados com base na morfologia da colônia. A unidade formadora de colônia mostrou E. coli como bactéria frequentemente encontrada no conteúdo fecal, oral e intestinal de todos os faisões acima. A diferença de significância geral foi encontrada entre as espécies bacterianas de faisões dourados, faisões verdes, faisões de pescoço anelado e faisões prateados. Verificou-se que a E.coli é predominantemente isolada de faisões saudáveis, seguida por Campylobacter, Staphylococcus e Brachyspira.

Isolation and characterization of bacteria residing in the oral, gut, and fecal samples of different pheasant species

Abstract There is a paucity of research conducted on microbial prevalence in pheasants. The microbiota of captive birds has zoonotic significance and must be characterize. Present study is therefore planned to assess the microbiota from oral, fecal and gut content of captive avian species. It will be helpful in characterization of harmful microbes. Different samples taken from oral, gut and feces of ring-necked pheasants (Phasianus colchicus), green pheasants (Phasianus versicolor), golden pheasant (Chrysolophus pictus) and silver pheasant (Lophura nycthemera). Samples were collected, diluted, and inoculated onto different agar plates (MacConkey, SS agar, MSA and nutrient agar) for cultivation of bacterial species. Colonies of E.coli, Staphylococcus spp. Brachyspira spp. and Campylobacter spp were observed based on colony morphology. Colony forming unit showed E. coli as frequently found bacteria in fecal, oral and gut contents of all the above pheasants. The overall significance difference was found among bacterial species of golden pheasants, green pheasant, ring-necked pheasant, and silver pheasants. It was concluded that E.coli is predominant isolated from heathy pheasants followed by Campylobacter, Staphylococcus and Brachyspira.

Resumo Há uma escassez de pesquisas realizadas sobre a prevalência microbiana em faisões. A microbiota de aves em cativeiro tem significado zoonótico e deve ser caracterizada. O presente estudo está, portanto, planejado para avaliar a microbiota do conteúdo oral, fecal e intestinal de espécies aviárias em cativeiro. Será útil na caracterização de micróbios nocivos. Diferentes amostras retiradas da boca, intestino e fezes de faisões de pescoço redondo (Phasianus colchicus), faisões verdes (Phasianus versicolor), faisões dourados (Chrysolophus pictus) e faisão prateado (Lophura nycthemera). As amostras foram coletadas, diluídas e inoculadas em diferentes placas de ágar (MacConkey, ágar SS, MSA e ágar nutriente) para o cultivo de espécies bacterianas. Colônias de E. coli, Staphylococcus spp., Brachyspira spp. e Campylobacter spp foram observados com base na morfologia da colônia. A unidade formadora de colônia mostrou E. coli como bactéria frequentemente encontrada no conteúdo fecal, oral e intestinal de todos os faisões acima. A diferença de significância geral foi encontrada entre as espécies bacterianas de faisões dourados, faisões verdes, faisões de pescoço anelado e faisões prateados. Verificou-se que a E.coli é predominantemente isolada de faisões saudáveis, seguida por Campylobacter, Staphylococcus e Brachyspira.

Isolation and characterization of bacteria residing in the oral, gut, and fecal samples of different pheasant species / Isolamento e caracterização de bactérias residentes nas amostras orais, intestinais e fecais de diferentes espécies de faisão

Abstract There is a paucity of research conducted on microbial prevalence in pheasants. The microbiota of captive birds has zoonotic significance and must be characterize. Present study is therefore planned to assess the microbiota from oral, fecal and gut content of captive avian species. It will be helpful in characterization of harmful microbes. Different samples taken from oral, gut and feces of ring-necked pheasants (Phasianus colchicus), green pheasants (Phasianus versicolor), golden pheasant (Chrysolophus pictus) and silver pheasant (Lophura nycthemera). Samples were collected, diluted, and inoculated onto different agar plates (MacConkey, SS agar, MSA and nutrient agar) for cultivation of bacterial species. Colonies of E.coli, Staphylococcus spp. Brachyspira spp. and Campylobacter spp were observed based on colony morphology. Colony forming unit showed E. coli as frequently found bacteria in fecal, oral and gut contents of all the above pheasants. The overall significance difference was found among bacterial species of golden pheasants, green pheasant, ring-necked pheasant, and silver pheasants. It was concluded that E.coli is predominant isolated from heathy pheasants followed by Campylobacter, Staphylococcus and Brachyspira.

Resumo Há uma escassez de pesquisas realizadas sobre a prevalência microbiana em faisões. A microbiota de aves em cativeiro tem significado zoonótico e deve ser caracterizada. O presente estudo está, portanto, planejado para avaliar a microbiota do conteúdo oral, fecal e intestinal de espécies aviárias em cativeiro. Será útil na caracterização de micróbios nocivos. Diferentes amostras retiradas da boca, intestino e fezes de faisões de pescoço redondo (Phasianus colchicus), faisões verdes (Phasianus versicolor), faisões dourados (Chrysolophus pictus) e faisão prateado (Lophura nycthemera). As amostras foram coletadas, diluídas e inoculadas em diferentes placas de ágar (MacConkey, ágar SS, MSA e ágar nutriente) para o cultivo de espécies bacterianas. Colônias de E. coli, Staphylococcus spp., Brachyspira spp. e Campylobacter spp foram observados com base na morfologia da colônia. A unidade formadora de colônia mostrou E. coli como bactéria frequentemente encontrada no conteúdo fecal, oral e intestinal de todos os faisões acima. A diferença de significância geral foi encontrada entre as espécies bacterianas de faisões dourados, faisões verdes, faisões de pescoço anelado e faisões prateados. Verificou-se que a E.coli é predominantemente isolada de faisões saudáveis, seguida por Campylobacter, Staphylococcus e Brachyspira.

Partial purification and biochemical characterization of an alkaline esterase from Sorghumbicolor

Esterases are enzymes that present good potential for industrial applications since they catalyze the formation or cleavage of ester bonds in water-soluble substrates, and sorghumseeds can represent an alternative source of this enzyme. The extraction of esterase from sorghumseeds is an economical alternative to obtain an enzyme of great interest. Esterases may improve the quality or accelerate the maturation of cheeses, cured bacon and fermented sausages and may also resolve racemic mixtures. Recently, seed esterases have been the focus of much attention as biocatalysts. In some cases, these enzymes present advantages over animal and microbial lipases due to some quite interesting features such as specificity and low cost, being a great alternative for their commercial exploitation as industrial enzymes The esterase studied here was extracted from sorghumseeds and some of its biochemical properties determined using synthetic substrates (p-nitrophenyl butyrate, caprylate, laurate and palmitate). The enzyme presented optimum activity at pH 8.0 and was stable in all the pH ranges studied. The optimum temperature for its activity was 40ºC but it showed low stability at this temperature (40% relative activity). The values derived for Km and Vmax were 0.67mM and 125 U.mg-1, respectively, obtained using p-nitrophenyl butyrate as the substrate. The enzyme showed an increase in activity when K2HPO4was added to the reaction medium, but the ions Mn2+, CO+, Hg+and Fe2+strongly inhibited the enzyme activity. This enzyme showed a preference for the hydrolysis of short chain fatty acids. The characteristics of sorghumesterase are very similar to those of the microbial esterases used in detergent processing.

Screening of bacterial extracellular xylanase producers with potential for cellulose pulp biobleaching

In this study, two hundred fifty-seven bacterial isolates from a suppressive soil library were screened to study their secretion of alkali-thermostable xylanases for potential use in cellulose pulp biobleaching. Xylanase activity was evaluated in solid and liquid media using xylan as the carbon source. Isolates were initially evaluated for the degradation of xylan in solid media by the congo red test. Selected strains were evaluated in liquid media for enzymatic activity and determination of total protein concentration using a crude protein extract (CPE). An isolate identified as Bacillus species TC-DT13 produced the highest amount of xylanase (1808 U mL-1). The isolate was active and stable at 70°C and pH 9.0, conditions which are necessary for the paper industry. This isolate can grow and produce xylanase in medium containing wheat fiber as a substrate. The CPE of this isolate was used in preliminary testing on cellulose pulp bleaching; enzyme treatment of the pulp resulted in a 5% increase of whiteness.

Relative Analysis of Gallic-Acid, Theobromine, Theophylline, and Caffeine Content among 12 Bangladeshi Tea Genotypes Using Reversed-Phase Liquid Chromatography

Aims: The tea industry is of great economic importance worldwide, owing to its possession of both food and medicinal values. Bangladesh is among the world-renowned tea exporting countries. However, the inadequate biochemical data for most cultivated Bangladeshi tea genotypes hinders its competitiveness on the world market. This is as a result of previous research mainly revolving around conventional breeding, mutagenesis, and polyploidization. This research aims to characterise the 12 Bangladeshi tea genotypes according to their biochemical content. Such information is inevitable in driving the demand and preference of these tea products on the world market. Study Design: The study was designed based on relevant research articles and standard laboratory procedures. Place and Duration of Study: This research was conducted at the Molecular Biology and Protein Science Laboratory, Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh, from June 2015 to September 2016. Methodology: Reversed-Phase Liquid Chromatography was used to determine the composition of different tea genotypes. A mixture of 999 ml de-ionised water & 1 ml TFA was used as buffer A, and 80% acetonitrile was used as buffer B in RPLC system. Results: We found that all the 12 tea genotypes are rich in Theophylline, Theobromine, Gallic-Acid, and Caffeine content, but with varying quantities. Conclusion: These results indicate that some of these tea genotypes can be used to produce the decaffeinated tea, a newly introduced tea product on the market that is on high demand. To ascertain the diversity of chemical composition among the various tea genotypes, biochemical characterisation of other Bangladeshi tea genotypes should be performed. Such data will enhance the market value and demand for Bangladeshi tea on the world market.

Thermostable chitinase from Cohnella sp. A01: isolation and product optimization

Abstract Twelve bacterial strains isolated from shrimp farming ponds were screened for their growth activity on chitin as the sole carbon source. The highly chitinolytic bacterial strain was detected by qualitative cup plate assay and tentatively identified to be Cohnella sp. A01 based on 16S rDNA sequencing and by matching the key morphological, physiological, and biochemical characteristics. The cultivation of Cohnella sp. A01 in the suitable liquid medium resulted in the production of high levels of enzyme. The colloidal chitin, peptone, and K2HPO4 represented the best carbon, nitrogen, and phosphorus sources, respectively. Enzyme production by Cohnella sp. A01 was optimized by the Taguchi method. Our results demonstrated that inoculation amount and temperature of incubation were the most significant factors influencing chitinase production. From the tested values, the best pH/temperature was obtained at pH 5 and 70 °C, with Km and V max values of chitinase to be 5.6 mg/mL and 0.87 µmol/min, respectively. Ag+, Co2+, iodoacetamide, and iodoacetic acid inhibited the enzyme activity, whereas Mn2+, Cu2+, Tweens (20 and 80), Triton X-100, and EDTA increased the same. In addition, the study of the morphological alteration of chitin treated by enzyme by SEM revealed cracks and pores on the chitin surface, indicating a potential application of this enzyme in several industries.

Production and partial characterization of proteases from Mucor hiemalis URM3773 / Produção e caracterização parcial de proteases a partir de Mucor hiemalis URM3773

The current study evaluated the proteases production from 11 fungal species belonging to the genera Mucor, Rhizomucor and Absidia. The species were obtained from the Collection of Cultures URM at the Mycology Department-UFPE, Brazil. The best producing species was Mucor hiemalis URM 3773 (1.689 U mL-1). Plackett-Burman design methodology was employed to select the most effective parameter for protease production out of 11 medium components, including: concentration of filtrate soybean, glucose, incubation period, yeast extract, tryptone, pH, aeration, rotation, NH4Cl, MgSO4 and K2HPO4. Filtrated soybean concentration was the significant variable over the response variable, which was the specific protease activity. The crude enzyme extract showed optimal activity in pH 7.5 and at 50ºC. The enzyme was stable within a wide pH range from 5.8 to 8.0, in the phosphate buffer 0.1M and in stable temperature variation of 40-70ºC, for 180 minutes. The ions FeSO4, NaCl, MnCl2, MgCl2 and KCl stimulated the protease activity, whereas ZnCl2 ion inhibited the activity in 2.27%. Iodoacetic acid at 1mM was the proteases inhibitor that presented greater action.The results indicate that the studied enzyme have great potential for industrial application.

Foi avaliada a produção de proteases por 11 espécies fúngicas pertencentes aos gêneros Mucor, Rhizomucor e Absidia, obtidas da Coleção de Culturas URM do Departamento de Micologia- UFPE, Brasil. A melhor espécie produtora foi Mucor hiemalis URM3773 (1,689 U mL-1). A metodologia de planejamento Plackett-Burman foi empregada para selecionar o parâmetro mais efetivo para a produção de proteases através de 11 componentes do meio, incluindo: concentração do filtrado de soja, glicose, período de incubação, extrato de levedura, triptona, pH, aeração, rotação, NH4Cl, MgSO4 e K2HPO4. A variável significante sobre a variável- resposta, atividade proteásica específica, foi a concentração do filtrado de soja. O extrato enzimático bruto apresentou atividade ótima ao pH 7,5 a 50ºC. A enzima foi estável em uma ampla variação de pH de 5,8­8,0 em tampão fosfato 0,1M e termicamente estável a uma variação de 40-70°C, durante 180 minutos. Os íons FeSO4, NaCl, MnCl2, MgCl2 e KCl estimularam a atividade proteásica, enquanto que o íon ZnCl 2 inibiu 2,27% da atividade. O inibidor de proteases que teve maior ação foi o ácido iodoacético a 1mM. Os resultados obtidos indicam que a enzima estudada tem grande potencial de aplicação industrial.

Different Methods for DNA Extraction from Yeast-Candida famata Isolated from Toddy.

Aims: Isolation and biochemical characterization of yeasts from toddy and standardization of best method for DNA extraction from yeast. Study Design: Biochemical characterization of yeast and genomic DNA extraction by manual and kits methods. Place and Duration of Study: Department of Microbiology, Institute of Genetic Engineering, Badu, kol-128, India and Molecular Mycopathology Lab, P. G. Department of Botany, Ramakrishna Mission Vivekananda Centenary College, Rahara, kol-118, India, from November 2012 –April,2013. Methodology: Toddy was collected in sterilized polythene bags from palm tree (Borassus flabellifer L; Family: Arecacea) in the morning, from Badu, 24-parganas (N) India. Isolation of yeasts was done by the method of Beech and Davenport [15] using MA (Malt extract) medium. Biochemical Identification was performed by using basal medium and procedure [1,2,15]. Genomic DNA extraction was done by manual and kits methods (UniflexTM DNA isolation Kit). Quality of extracted DNA was checked by the absorbance ratio (A260 / A280) ranged from 1.8 to 2.0. Results: By performing morphological, microscopical and biochemical characterization the isolated yeast from toddy was identified as Candida famata consulting with the key of yeast published [1,2]. The UniflexTM DNA isolation Kit method is much more convenient way to get pure and high quality DNA than the manual methods. Conclusion: Isolated yeast from toddy was identified as Candida famata. The genomic DNA of Candida famata was extracted purely by UniflexTM DNA isolation Kit. This method was better and more convenient than manual method.

Caracterización de los tipos capsulares de Pasteurella multocida en exudado faríngeo de bovinos productores de carne clínicamente sanos en el estado de Querétaro / Characterization of capsular types of Pasteurella multocida isolated from clinically healthy beef cattle pharyngeal exudate in the state of Querétaro

El objetivo del presente estudio fue identificar y caracterizar los tipos capsulares de P. multocida en exudado faríngeo en bovinos destinados a la producción de carne en el estado de Querétaro. Se obtuvieron, mediante hisopo, 227 muestras de exudado faríngeo de animales clínicamente sanos en una planta de sacrificio ubicada en el municipio de Ezequiel Montes, Querétaro. Las muestras se sembraron en agar sangre y se incubaron a 37°C por 24 h en aerobiosis. Las cepas aisladas fueron identificadas mediante características morfológicas, pruebas bioquímicas convencionales y el microsistema comercial API 20NE. La tipificación de los grupos capsulares A y D se realizó por medio de una PCR múltiple para la amplificación de los genes hyaD-hyaC y dcbF, respectivamente. De acuerdo con los valores establecidos por el software API WEB, se logró la identificación de 14.09% (32/227) de cepas de P. multocida, que mostraron 96% de identidad y una tipicidad de 1 a P. multocida. Por medio de la PCR múltiple se logró la amplificación de los genes hyaD-hyaC correspondientes al grupo capsular A en el 100% (32/32) de las cepas identificadas previamente como P. multocida. No existen datos similares en México sobre la identificación y caracterización de P. multocida en bovinos destinados a la producción de carne. Con los resultados obtenidos se corrobora que, de manera similar a otros países de Europa y América, en México el grupo capsular predominante de P. multocida es el A.

The objective of the present study was to identify and characterize capsular types of P. multocida isolated from beef cattle pharyngeal exudate in the state of Querétaro. Two hundred and twenty seven pharyngeal exudate swab samples from clinically healthy animals in a slaughterhouse in the municipality of Ezequiel Montes, Querétaro were obtained. Samples were seeded in blood agar and incubated at 37°C for 24 h under aerobiosis. Strains were identified through morphological characteristics, conventional biochemical tests and commercial API 20NE Micro-System. Capsular typing of groups A and D was performed by a multiplex PCR for amplification of genes hyaD-hyaC and dcbF, respectively. According to the values established by API WEB software, it was possible to identify 14.09% (32/227) of P. multocida strains, which showed an identification percentage of 96% and a typicality of 1 to P. multocida. By multiplex PCR, the amplification of genes hyaD-hyaC, correspondent to capsular group A in 100% (32/32) of the strains previously identified as P. multocida, was achieved. There are no similar data in Mexico on the identification and characterization of P. multocida in beef cattle. With the results obtained it is confirmed that, in a similar way with other countries of Europe and America, capsular type A of P. multocida is predominant in Mexico.

Cytokinin Dehydrogenase activity in primary roots and characterization of primary metabolites from leaves and rootlets of Ricinus Communis

The aim of this study was to determine the activity of cytokinin dehydrogenase (CKX) and to measure other biochemical components in the primary leaves and radicles of castor seedlings (BRS Energia) in the initial phase of growth. The crude protein extract obtained after a 1-h extraction from the root tissues of seedlings showed no detectable CKX enzymatic activity when incubated with the substrate isopentenyl adenine for 1 h. However, after precipitation with ammonium sulfate at 70% saturation, the pellet showed CKX activity. The peroxidase enzyme activity was higher in the leaves than in the radicles. The total and reducing sugar content was 1.5 times higher in the leaves than in the radicles. The amino acid and protein contents were 6.4 and 9.2 times higher in the leaves than in the radicles, respectively.

Isolation of Microorganisms from Dairy Effluent.

Aim: Every Dairy industry has problems of effluent treatment. This can be revealed by effective treatment of the effluent. The effective treatment can be done by using microorganisms to stabilize the organic and inorganic load of the effluent. The aim of the present work is to study the dairy wastewater micro biota and to identify some new active strains which can bring about fast biodegradation of the organic compounds. Study Design: Isolation and determination of bacteriological characteristics of the dairy effluents. Methodology: Studies were carried out to isolate the microorganisms from collected effluent sample from the dairies under studies. Isolation of microorganisms was done by primary screening, Cultural Characterization, Biochemical characterization and Identified by using Bergey’s Manuals of Systematic Bacteriology. Results: During 2011-2013 from two different districts of Maharashtra (India), dairy industry effluents were collected for the isolation of micro organisms. Effluent samples were collected as per Jacksch and piper method and primary screening was done and totally 7 Isolates were screened out. These isolates were characterized on nutrient agar at room temperature for 24 hrs. Isolates were observed for the cultural characters like size, shape, colour, margin, elevation, opacity and consistency and morphological characters like Gram nature, sporulation, shape and arrangement of cells, motility etc. and were recorded. Physio-biochemical characterization was followed by biochemical tests for enzymatic activities like catalase, oxidase, nitrate reduction, urease, caseinase etc and carbohydrates utilization tests for lactose, maltose, inositol, xylose etc. performed to check their ability for metabolization. On the basis of these characteristics, isolates were identified by using Bergey’s Manuals of Systematic Bacteriology. The identified Bacterial Isolates were of Genus Lactobacillus, Bacillus, Staphylococcus, Enterococcus, Listeria etc. Conclusion: These Bacterial isolates have the ability to utilize the components like nitrate, starch, gelatin, sugars like sucrose, maltose, lactose etc. which was confirmed by the biochemical tests. Bacterial flora from the effluents can be identified and efficiently applied for the biological treatment of the dairy effluents.

Biochemical characterisation of a glucoamylase from Aspergillus niger produced by solid-state fermentation

In this work, glucoamylase was produced by Aspergillus niger in solid-state fermentation. The enzyme was partially purified by ammonium sulphate precipitation and ion exchange and gel filtration chromatographies. Its molecular mass was estimated as 118.17 kDa by electrophoresis. The partially purified enzyme had an optimum pH range of 4.5-5.0 and an optimum temperature of 60 °C, with average activity 152.85 U mL-1. Thermal and pH stability assays with the crude extract showed that more than 60 percent of the activity remained at pH 4.6 and 60 °C, even after an exposition to these conditions longer than 24 h. Yet, after purification, the enzyme was stable at these for at least 4 h, which indicated that its purification for use in starch saccharification was inadvisable. K M and Vmax were 0.34 mg mL-1 and 160.22 U mL-1, respectively.

Biochemical characterization of embryonic chick calvarial cells / 대한치과교정학회지

Chicken calvarial bone is known to contain various cell types, but their exact composition is unknown. By characterizing the chicken calvarial bone biochemically, it can be used to study biochemical, histochemical actions of bone cells in general. Calvaria of I8-day-old white leg horn embryo was aseptically dissected and bone cell populations were isolated by sequential enzymatic digestion. Histochemical study for osteoclast-like bone cell population was performed with tartrate resistant acid phosphatase(TRAP) stain and for osteoblast-like bone cell population, alkaline phosphatase(ALP) stain was performed. Biochemical study for osteoblast-like bone cell population was performed using alkaline phosphatasetal.P) assay. Following conclusions were obtained from this study. 1. TRAP positive multi and mononuclear cells were mostly observed in group I and II, indicating that osteoclast-like bone cell population is mostly found in these groups. 2. All the cultured groups showed almost equal ALP activities and were positive for ALP stain, indicating that osteoblast-like bone cell population is evenly dispersed in all culture groups. 3. Experimental group treated with I,25(OH)2D3 showed increase in ALP activity in contrast to the control group, confirming previous studies that I,25(OH)2D3 increases ALP activities in vitro bone cultures. 4. Results from von Kassa's stain indicated that in vitro bone formation had occured after 3 weeks of culture with beta glycero phosphate.