Population and genetic diversity of rhizobia nodulating chickpea in Indo-Gangetic plains of India.

Chickpea is a crucial leguminous crop and India is the leading producer, with an average yield of 1.18 tons/ha. It is renowned for its specific nodulation with rhizobia. Despite its significance, studies on chickpea-nodulating rhizobia often focused on small-scale investigations within restricted geographical areas. This study delves into the population, genetic diversity, and symbiotic efficiency of chickpea-nodulating rhizobia in the Indo-Gangetic Plains (IGP) of India. The study revealed a low population of chickpea rhizobia (ranging from 11 to 565 cells/g dry soil) across the examined area. Only three samples exhibited a population exceeding 300 cells/g, emphasizing the potential need for inoculation of rhizobia with efficient and competitive strains. Correlation analysis highlighted a significant positive correlation between rhizobial population and organic carbon content, among various soil parameters like pH, electrical conductivity, available nitrogen (N), phosphorus (P), potassium (K), and organic carbon content. Among the 79 presumptive rhizobia isolated from 24 IGP locations, 61 successfully nodulated chickpea cultivar Pusa 362. 16S rRNA gene sequencing categorized 54 isolates as Mesorhizobium, four as Rhizobium, and three as Ensifer. Genetic diversity assessed by BOX-PCR revealed sixteen distinct banding patterns, underscoring substantial variability among the strains. The strains exhibited plant growth-promoting activities, salt tolerance up to 3% NaCl, and pH tolerance between 4 and 10. Six symbiotically efficient strains were identified based on their positive impact on nodulation and dry biomass. This study provides crucial insights into the diversity, genetic makeup, and symbiotic efficiency of chickpea rhizobia in the IGP, supporting the potential use of indigenous rhizobia for sustainable chickpea productivity in the region.

Crop rotation and inoculation increase soil bradyrhizobia population, soybean grain yields, and profitability.

Crop rotation and rhizobial inoculation are strategies to increase yield by means of organic matter addition and modulation of microbial diversity. However, the extent to which these agricultural practices change soil Bradyrhizobium populations, soybean grain yield, and economic benefits to farmers is unclear. Thus, this study aimed to evaluate the interaction between crop rotation and inoculation of soybean (Glycine max) cultivated in two contrasting soils (clayey and sandy soil) on biological nitrogen fixation components, grain yields, and profits. Field experiments with a three-year crop rotation system were carried out to compare effects of inoculation and crop rotations on soil chemical attributes, bradyrhizobia most probable number (MPN) and diversity, soybean nodulation, grain yield, and economic indicators of inoculation in different crop rotations. The crop rotation did not affect the soil MPN cells of bradyrhizobia, but the inoculation and the soil sampling time did, ranging from 3.61-4.42 to 4.40-4.82 in the sandy soil, while in the clayey soil they were from 5.19-6.34 to 6.61-7.14 in Log10 per g of soil with higher population after harvest of summer crops. In the clayey soil, crop rotation influenced soybean nodulation. The grain yield of inoculated soybean in the clayey soil was higher than that in the sandy soil. Soybean inoculation with Bradyrhizobium spp. increased the profitability of agricultural production systems by up to 45% in clayey soil and up to 7% in sandy soil.

Occurrence of diverse Bradyrhizobium spp. in roots and rhizospheres of two commercial Brazilian sugarcane cultivars.

The genus Bradyrhizobium harbors many endosymbionts of legumes, but recent research has shown their widespread presence in soils and in non-legumes, notably in roots of sugarcane. This study aimed to investigate the Bradyrhizobium sp. community density in the endosphere and the rhizosphere of two commercial sugarcane cultivars. Samples of the rhizosphere and root endosphere of two Brazilian sugarcane cultivars (RB867515 and IACSP95-5000) were collected, serially diluted, and inoculated on axenic cowpea (Vigna unguiculata) and the induction of nodules was evaluated. Based on the results, a density was estimated of at least 1.6 × 104 rhizobia g root-1 in rhizosphere samples and up to 105 rhizobia g root -1 in endosphere. BOX-PCR profiling of 93 Bradyrhizobium isolates revealed genetic variability, with some dominant (up to 18 representants) and less dominant genotypes. 16S rRNA and ITS sequence analyses confirmed nine phylotypes, six of which pertained to the B. elkanii clade and three to the B. japonicum clade. Five isolates were genetically similar to the recently described species B. sacchari. There was no effect of the factors "plant cultivar" and "root compartment" on Bradyrhizobium sp. community composition and the most abundant genotypes occurred both in rhizosphere and endosphere of both cultivars. Therefore, this study confirms the natural presence of diverse Bradyrhizobium spp. in sugarcane root systems (mainly the rhizosphere) and indicates that certain Bradyrhizobium phylotypes have a special affinity for sugarcane root colonization.

Comparação de métodos para quantificação de bactérias nitrificantes / Comparison of methods for quantification of nitrifying bacteria

RESUMO A quantificação de bactérias nitrificantes é de extrema importância para o monitoramento de sistemas biológicos de tratamento que promovam a nitrificação. Neste trabalho, 15 amostras de efluentes coletadas em sistema de tratamento por lodos ativados (LA) foram analisadas de modo a quantificar bactérias nitrificantes por meio de duas técnicas: tubos múltiplos ou técnica do número mais provável (NMP); e hibridação in situ fluorescente (FISH). Os resultados sugerem que houve uma tendência de se obter valores diferentes para bactérias oxidadoras de amônia por meio da NMP em comparação com a FISH. Não obstante, a análise estatística revelou que a diferença de quantificação encontrada entre as técnicas não foi significativa, indicando que ambas podem ser usadas. Para as oxidadoras de nitrito, não foi possível realizar comparação, uma vez que os gêneros que estavam sendo determinados em cada uma das técnicas provavelmente eram diferentes. Sendo assim, as técnicas NMP e FISH se mostraram métodos relativamente simples e adequados para quantificação de microrganismos nitrificantes, com vantagens e limitações inerentes a cada uma.

ABSTRACT The quantification of nitrifying bacteria is of utmost importance for monitoring biological treatment systems designed to promote nitrification. In this study, 15 activated sludge samples were analyzed in order to quantify nitrifying bacteria by two different methods: the most-probable number (MPN); and the fluorescence in situ hybridization (FISH). The results suggest that there was a tendency to obtain different values for ammonia-oxidizing bacteria by MPN compared to FISH. However, statistical analysis of these data revealed that the difference found between the two techniques was not significant, indicating that both can be used for quantification of ammonia-oxidizing bacteria. For nitrite-oxidizing bacteria it was not possible to make this comparison, since the bacterial genera that were being determined in each technique were likely different. Thus, MPN and FISH techniques proved to be relatively simple and suitable for quantification of nitrifying microorganisms in sludge samples, each of them with advantages and limitations.

Quantification of Vibrio species in oysters from the Gulf of Mexico with two procedures based on MPN and PCR.

Oysters can accumulate potentially pathogenic water bacteria. The objective of this study was to compare two procedures to quantify Vibrio species present in oysters to determine the most sensitive method. We analyzed oyster samples from the Gulf of Mexico, commercialized in Mexico City. The samples were inoculated in tubes with alkaline peptone water (APW), based on three tubes and four dilutions (10-1 to 10-4). From these tubes, the first quantification of Vibrio species was performed (most probable number (MPN) from tubes) and bacteria were inoculated by streaking on thiosulfate-citrate-bile salts-sucrose (TCBS) petri dishes. Colonies were isolated for a second quantification (MPN from dishes). Polymerase chain reaction (PCR) was used to determine species with specific primers: ompW for Vibrio cholerae, tlh for Vibrio parahaemolyticus, and VvhA for Vibrio vulnificus. Simultaneously, the sanitary quality of oysters was determined. The quantification of V. parahaemolyticus was significantly higher in APW tubes than in TCBS dishes. Regarding V. vulnificus counts, the differences among both approaches were not significant. In contrast, the MPNs of V. cholerae obtained from dishes were higher than from tubes. The quantification of MPNs through PCR of V. parahaemolyticus and V. vulnificus obtained from APW was sensitive and recommendable for the detection of both species. In contrast, to quantify V. cholerae, it was necessary to isolate colonies on TCBS prior PCR. Culturing in APW at 42 °C could be an alternative to avoid colony isolation. The MPNs of V. cholerae from dishes was associated with the bad sanitary quality of the samples.

Quantitative distribution of Salmonella spp. and Escherichia coli on beef carcasses and raw beef at retail establishments.

Salmonella is a foodborne pathogen that commonly inhabits the gastrointestinal tract of a healthy feedlot cattle and can be transferred to the carcass surface during hide removal and evisceration procedures. Numerous investigations on Salmonella prevalence throughout different stages of the beef chain have been conducted. In contrast, limited studies are available on quantitative determinations of Salmonella at different steps in raw meat production. Quantitative data, particularly for pathogenic bacteria such as Salmonella are important for quantitative risk assessment. Salmonella spp. and Escherichia coli populations were enumerated on beef carcass samples collected at abattoirs and also in beef chunks and ground beef samples collected from butcher's shops at retail in Jalisco State, Mexico. Sponge samples from beef carcass sides (n=142) were collected immediately after final water wash and before chilling at three non-federally inspected abattoirs following USDA-FSIS sampling protocols. Beef chunks (n=84) and ground beef (n=65) samples were obtained from 86 butcher's shops. Salmonella enumeration was conducted by the Most Probable Number method and E. coli counts were determined using Petrifilm plates. Salmonella was isolated from 18% of beef carcasses, 39% of beef chunks and 71% of ground beef samples. Salmonella mean counts were 1.3±0.9 Log MPN/300 cm(2) on beef carcasses, 1.9±0.9 and 2.3±1.1 Log MPN/25 g in beef chunks and ground beef samples, respectively. Twenty-six Salmonella serotypes and 11 serogroups were identified among 432 isolates recovered. Salmonella typhimurium (14%), Salmonella sinstorf (12%) and S. Group E1 monophasic (10%) were the most frequent. Escherichia coli was present on 97, 84 and 100% of beef carcasses, beef chunks and ground beef samples, respectively. Escherichia coli mean counts were 3.2±0.7 Log CFU/300 cm(2), 3.9±1.1 and 4.5±1.2 Log CFU/25 g on beef carcasses, beef chunks and ground beef, respectively. Salmonella prevalence and mean counts found in raw beef were higher than previously reported in studies from other countries. The data collected in this study show a trend in the prevalence of Salmonella to be higher as meat processing is extended at retail. This, together with the diversity of serotypes found, indicates that raw meat is exposed to multiple contamination sources during slaughter and retail processing and highlights the necessity to implement Sanitation Standard Operating Procedures for those establishments. Finally, this study provides quantitative information for future risk assessments associated with the risk of human salmonellosis.

Número mais provável miniaturizado e microbiologia convencional para isolamento de Salmonella spp. em abatedouros de frangos de corte / Miniaturized most probable number and conventional microbiology for isolation of Salmonella spp. in poultry slaughterhouses

Os produtos de origem avícola podem ser importantes veículos de transmissão de Salmonella spp. para humanos e, dentre os vários parâmetros que determinam a qualidade de um alimento, destacam-se os que definem suas características microbiológicas. Objetivou-se detectar e quantificar Salmonella spp. na tecnologia de abate de frangos de corte por microbiologia convencional (MC) e número mais provável miniaturizado (mNMP). As coletas foram realizadas em duas visitas a três abatedouros sob Inspeção Federal e em seis pontos de coleta em triplicata, definidos como: recepção das aves (swabs de cloaca e esponjas de gaiolas de transporte antes e após a higienização) e carcaças (após pré resfriamento em chiller, após o gotejamento e antes da embalagem primária e congeladas a -12oC por 24 horas), totalizando 108 amostras. Identificou-se Salmonella spp. em três dos seis pontos do fluxograma de abate e em dois dos três estabelecimentos amostrados, independentemente do método utilizado, perfazendo 5,5% de positividade, onde destaca-se a contaminação nas gaiolas de transporte das aves após a higienização. Não foi possível correlacionar os resultados da microbiologia convencional e do mNMP ou mesmo quantificar a contaminação ao longo da tecnologia de abate, o que indica a necessidade de se utilizar um método qualitativo aliado ao método de quantificação quando Salmonella estiver presente em quantidades inferiores ao limite de detecção do mNMP proposto (0,13 NMP/mL). Os sorovares identificados foram Typhimurium, Panama, Lexington e Rissen, consideradas paratíficos e, portanto, potencialmente capazes de causar infecções em humanos, embora estes sorovares não tenham sido isolados em produtos finais e sim na chegada dos frangos aos abatedouros (swabs de cloaca e gaiolas de transporte). A identificação de Salmonella spp. nas gaiolas de transporte após a higienização é um indicativo da necessidade de revisão e adequação dos métodos automatizados de lavagem atualmente utilizados nos abatedouros.(AU)

Poultry products can be important modes of transmission of Salmonella spp. to humans and, among several parameters used to determine food quality, microbiological characteristics play an essential role. The aim of this study was to determine and quantify Salmonella spp. at broiler slaughtering facilities. This was done by conventional microbiology and by the miniaturized most probable number (mMPN) methods. Three federally-inspected slaughterhouses were visited, where samples were collected in triplicate from six sites: reception of live birds (cloacal swabs and sponge samples from transport cages before and after sanitation) and carcass processing (after pre-chiller, after dripping, and before primary packaging and refrigeration at -12oC for 24h), totaling 108 samples. Three of the six surveyed sites and two of the three slaughterhouses were contaminated with Salmonella spp., showing an infection rate of 5.5% independently of the method used, and revealing that transport cages were contaminated after sanitation. No correlation could be established between the results of conventional microbiology and mMPN methods, and contamination along the slaughtering line could not quantified. This indicates the importance of combining qualitative and quantitative methods for the enumeration of Salmonella when detection rates are lower than the proposed mMPN limit (0.13 MPN/mL). Typhimurium, Panama, Lexington and Rissen, which are paratyphoid organisms and are potentially infectious to humans, were identified. However, these serovars were isolated at the reception of live birds (from cloacal swabs and from transport cages) rather than from the end products. Given that Salmonella spp. was detected in transport cages after sanitation, it is paramount that automated washing procedures currently used in slaughterhouses be reassessed and adjusted.(AU)

Número mais provável miniaturizado e microbiologia convencional para isolamento de Salmonella spp. em abatedouros de frangos de corte / Miniaturized most probable number and conventional microbiology for isolation of Salmonella spp. in poultry slaughterhouses

Os produtos de origem avícola podem ser importantes veículos de transmissão de Salmonella spp. para humanos e, dentre os vários parâmetros que determinam a qualidade de um alimento, destacam-se os que definem suas características microbiológicas. Objetivou-se detectar e quantificar Salmonella spp. na tecnologia de abate de frangos de corte por microbiologia convencional (MC) e número mais provável miniaturizado (mNMP). As coletas foram realizadas em duas visitas a três abatedouros sob Inspeção Federal e em seis pontos de coleta em triplicata, definidos como: recepção das aves (swabs de cloaca e esponjas de gaiolas de transporte antes e após a higienização) e carcaças (após pré resfriamento em chiller, após o gotejamento e antes da embalagem primária e congeladas a -12oC por 24 horas), totalizando 108 amostras...

Poultry products can be important modes of transmission of Salmonella spp. to humans and, among several parameters used to determine food quality, microbiological characteristics play an essential role. The aim of this study was to determine and quantify Salmonella spp. at broiler slaughtering facilities. This was done by conventional microbiology and by the miniaturized most probable number (mMPN) methods. Three federally-inspected slaughterhouses were visited, where samples were collected in triplicate from six sites: reception of live birds (cloacal swabs and sponge samples from transport cages before and after sanitation) and carcass processing (after pre-chiller, after dripping, and before primary packaging and refrigeration at -12oC for 24h), totaling 108 samples...

Miniaturized most probable number for the enumeration of Salmonella sp in artificially contaminated chicken meat

Salmonella is traditionally identified by conventional microbiological tests, but the enumeration of this bacterium is not used on a routine basis. Methods such as the most probable number (MPN), which utilize an array of multiple tubes, are time-consuming and expensive, whereas miniaturized most probable number (mMPN) methods, which use microplates, can be adapted for the enumeration of bacteria, saving up time and materials. The aim of the present paper is to assess two mMPN methods for the enumeration of Salmonella sp in artificially-contaminated chicken meat samples. Microplates containing 24 wells (method A) and 96 wells (method B), both with peptone water as pre-enrichment medium and modified semi-solid Rappaport-Vassiliadis (MSRV) as selective enrichment medium, were used. The meat matrix consisted of 25g of autoclaved ground chicken breast contaminated with dilutions of up to 10(6) of Salmonella Typhimurium (ST) and Escherichia coli (EC). In method A, the dilution 10-5 of Salmonella Typhimurium corresponded to >57 MPN/mL and the dilution 10-6 was equal to 30 MPN/mL. There was a correlation between the counts used for the artificial contamination of the samples and those recovered by mMPN, indicating that the method A was sensitive for the enumeration of different levels of contamination of the meat matrix. In method B, there was no correlation between the inoculated dilutions and the mMPN results.(AU)

Miniaturized most probable number for the enumeration of Salmonella sp in artificially contaminated chicken meat

Salmonella is traditionally identified by conventional microbiological tests, but the enumeration of this bacterium is not used on a routine basis. Methods such as the most probable number (MPN), which utilize an array of multiple tubes, are time-consuming and expensive, whereas miniaturized most probable number (mMPN) methods, which use microplates, can be adapted for the enumeration of bacteria, saving up time and materials. The aim of the present paper is to assess two mMPN methods for the enumeration of Salmonella sp in artificially-contaminated chicken meat samples. Microplates containing 24 wells (method A) and 96 wells (method B), both with peptone water as pre-enrichment medium and modified semi-solid Rappaport-Vassiliadis (MSRV) as selective enrichment medium, were used. The meat matrix consisted of 25g of autoclaved ground chicken breast contaminated with dilutions of up to 10(6) of Salmonella Typhimurium (ST) and Escherichia coli (EC). In method A, the dilution 10-5 of Salmonella Typhimurium corresponded to >57 MPN/mL and the dilution 10-6 was equal to 30 MPN/mL. There was a correlation between the counts used for the artificial contamination of the samples and those recovered by mMPN, indicating that the method A was sensitive for the enumeration of different levels of contamination of the meat matrix. In method B, there was no correlation between the inoculated dilutions and the mMPN results.

Miniaturized most probable number for the enumeration of Salmonella sp in artificially contaminated chicken meat

Salmonella is traditionally identified by conventional microbiological tests, but the enumeration of this bacterium is not used on a routine basis. Methods such as the most probable number (MPN), which utilize an array of multiple tubes, are time-consuming and expensive, whereas miniaturized most probable number (mMPN) methods, which use microplates, can be adapted for the enumeration of bacteria, saving up time and materials. The aim of the present paper is to assess two mMPN methods for the enumeration of Salmonella sp in artificially-contaminated chicken meat samples. Microplates containing 24 wells (method A) and 96 wells (method B), both with peptone water as pre-enrichment medium and modified semi-solid Rappaport-Vassiliadis (MSRV) as selective enrichment medium, were used. The meat matrix consisted of 25g of autoclaved ground chicken breast contaminated with dilutions of up to 10(6) of Salmonella Typhimurium (ST) and Escherichia coli (EC). In method A, the dilution 10-5 of Salmonella Typhimurium corresponded to >57 MPN/mL and the dilution 10-6 was equal to 30 MPN/mL. There was a correlation between the counts used for the artificial contamination of the samples and those recovered by mMPN, indicating that the method A was sensitive for the enumeration of different levels of contamination of the meat matrix. In method B, there was no correlation between the inoculated dilutions and the mMPN results.

Prevalence and quantification of Listeria monocytogenes in beef offal at retail level in Selangor, Malaysia

A total of 63 beef offal samples (beef liver = 16; beef lung = 14; beef intestine = 9; beef tripe = 15; beef spleen = 9) from three wet markets (A, B, and C) in Selangor, Malaysia were examined for the prevalence and microbial load of Listeria monocytogenes. A combination of the most probable number and polymerase chain reaction (MPN-PCR) method was employed in this study. It was found that L. monocytogenes detected in 33.33% of the beef offal samples. The prevalence of L. monocytogenes in beef offal purchased from wet markets A, B, and C were 22.73%, 37.50% and 41.18% respectively. The density of L. monocytogenes in all the samples ranged from<3upto> 2,400 MPN/g. The findings in this study indicate that beef offal can be a potential vehicle of foodborne listeriosis.

Dinâmica de rizóbios em solo do cerrado de Roraima durante o período de estiagem / Dynamic of rhizobia in the soil during the dry season in cerrado of Roraima

The biological nitrogen fixation in legumes is performed by a group of bacteria known as rhizobia. The survival of these bacteria in soils is affected by several factors, such as temperature, drought and soil fertility. This study was performed to evaluate the dynamics of rhizobia in the soil after soybean cultivation and during a dry season in the cerrado of Roraima. Three areas were sampled: i) native cerrado as reference; ii) an area previously cultivated with soybean for one season; and iii) another one cultivated for two seasons also with soybean. The soil was sampled at a depth of 0-10 cm in five times (0, 45, 90, 135 and 180 days) during the dry season (September 2006 to March 2007). The rhizobial density in the soil was evaluated by the most probable number method with infection of soybean and cowpea plants. It was observed very low number of soybean nodulating bacteria in the reference area, but a high density, of up to several hundred rhizobia capable to nodulate cowpea was measured in this same area. Cropping of soybean with inoculated seeds increased rhizobial density evaluated by both trapping hosts. In cropped areas, an intense reduction of rhizobium density was observed just after soybean harvest, and this reduction continued until the end of the period of evaluation. It was concluded that soybean cultivation increases the density of rhizobial in the cerrado soil; however, this density is drastically reduced, during the dry season, by 99% at the end of the dry period.(AU)

A fixação biológica de nitrogênio que ocorre em leguminosas é realizada por um grupo de bactérias conhecidas como rizóbios. A sobrevivência destas bactérias no solo é influenciada por diversos fatores como a temperatura, umidade e fertilidade do solo. O objetivo deste trabalho foi avaliar a dinâmica da população de rizóbios em solo após o cultivo de soja, durante o período de estiagem no cerrado de Roraima. Foram amostradas três áreas: i) cerrado nativo como referência; ii) área cultivada uma vez com soja inoculada com rizóbio; iii) e área cultivada duas vezes com soja inoculada com rizóbio em anos consecutivos. O solo foi coletado na profundidade de 0-10 cm em cinco períodos a partir do inicio da estiagem no mês de setembro de 2006 coincidindo com a época de colheita da soja e prolongando-se até março de 2007 (0, 45, 90, 135 e 180 dias). A população de rizóbios no solo foi avaliada pela técnica do número mais provável (NMP) utilizando plantas de soja e de feijão-caupi como espécies isca. Foi observado que na área nativa praticamente não existiam bactérias nodulantes de soja, mas havia uma população capaz de nodular o feijão-caupi de até algumas centenas de rizóbios por gramas de solo. O cultivo da soja utilizando sementes inoculadas elevou a população de rizóbios no solo que foi constatada por ambas às espécies de plantas isca. Nas áreas cultivadas, constatou-se uma intensa redução da população de rizóbios no solo, em especial logo após a colheita da soja, continuando o decréscimo até o último período de avaliação. Conclui-se que o cultivo da soja inoculada com rizóbio eleva a densidade de rizóbios em solo do cerrado, mas durante a estiagem ocorre uma drástica redução dessa população, que pode chegar a mais de 99% considerando o início e final do período.(AU)

Dinâmica de rizóbios em solo do cerrado de Roraima durante o período de estiagem / Dynamic of rhizobia in the soil during the dry season in cerrado of Roraima

The biological nitrogen fixation in legumes is performed by a group of bacteria known as rhizobia. The survival of these bacteria in soils is affected by several factors, such as temperature, drought and soil fertility. This study was performed to evaluate the dynamics of rhizobia in the soil after soybean cultivation and during a dry season in the cerrado of Roraima. Three areas were sampled: i) native cerrado as reference; ii) an area previously cultivated with soybean for one season; and iii) another one cultivated for two seasons also with soybean. The soil was sampled at a depth of 0-10 cm in five times (0, 45, 90, 135 and 180 days) during the dry season (September 2006 to March 2007). The rhizobial density in the soil was evaluated by the most probable number method with infection of soybean and cowpea plants. It was observed very low number of soybean nodulating bacteria in the reference area, but a high density, of up to several hundred rhizobia capable to nodulate cowpea was measured in this same area. Cropping of soybean with inoculated seeds increased rhizobial density evaluated by both trapping hosts. In cropped areas, an intense reduction of rhizobium density was observed just after soybean harvest, and this reduction continued until the end of the period of evaluation. It was concluded that soybean cultivation increases the density of rhizobial in the cerrado soil; however, this density is drastically reduced, during the dry season, by 99% at the end of the dry period.

A fixação biológica de nitrogênio que ocorre em leguminosas é realizada por um grupo de bactérias conhecidas como rizóbios. A sobrevivência destas bactérias no solo é influenciada por diversos fatores como a temperatura, umidade e fertilidade do solo. O objetivo deste trabalho foi avaliar a dinâmica da população de rizóbios em solo após o cultivo de soja, durante o período de estiagem no cerrado de Roraima. Foram amostradas três áreas: i) cerrado nativo como referência; ii) área cultivada uma vez com soja inoculada com rizóbio; iii) e área cultivada duas vezes com soja inoculada com rizóbio em anos consecutivos. O solo foi coletado na profundidade de 0-10 cm em cinco períodos a partir do inicio da estiagem no mês de setembro de 2006 coincidindo com a época de colheita da soja e prolongando-se até março de 2007 (0, 45, 90, 135 e 180 dias). A população de rizóbios no solo foi avaliada pela técnica do número mais provável (NMP) utilizando plantas de soja e de feijão-caupi como espécies isca. Foi observado que na área nativa praticamente não existiam bactérias nodulantes de soja, mas havia uma população capaz de nodular o feijão-caupi de até algumas centenas de rizóbios por gramas de solo. O cultivo da soja utilizando sementes inoculadas elevou a população de rizóbios no solo que foi constatada por ambas às espécies de plantas isca. Nas áreas cultivadas, constatou-se uma intensa redução da população de rizóbios no solo, em especial logo após a colheita da soja, continuando o decréscimo até o último período de avaliação. Conclui-se que o cultivo da soja inoculada com rizóbio eleva a densidade de rizóbios em solo do cerrado, mas durante a estiagem ocorre uma drástica redução dessa população, que pode chegar a mais de 99% considerando o início e final do período.

Calidad bacteriológica de aguas en piscinas públicas y privadas de la ciudad de Cumaná, estado Sucre, Venezuela / Bacteriological quality of water of public and private pools of the city of Cumaná, Sucre state, Venezuela

Para evaluar la calidad bacteriológica de aguas de piscinas en la ciudad de Cumaná, estado Sucre, Venezuela, se recolectaron muestras de agua en 1 piscina pública y 4 privadas, codificadas de la A a la E; se tomaron 2 muestras semanales durante 2 meses, antes y después de la limpieza. Se determinó pH, temperatura y cloro residual; los aerobios mesófilos por contaje en placas, el Número Más Probable (NMP) para coliformes totales (CT) y fecales (CF) e identificación bacteriana por métodos convencionales. El pH osciló entre 6,8 y 7,3, la temperatura de 29 a 31ºC y el cloro residual de 0,3 a 0,5 mg/L. El contaje más elevado de bacterias mesófilas se obtuvo en B con 6x10 2 UFC/mL, y el más bajo en C con 3x10 2 UFC/mL. En relación al NMP, antes de la limpieza, el valor más alto se obtuvo en D con 2,8x10 3 CT/100 ml; E mostró el valor más alto de CF /100 ml. Después de la limpieza B mostró el valor más alto de CT ubicándose en 9,3mLx10 2 y los valores más alto de CF para D y E en 3x10 2 . Los valores de CF antes y después de la limpieza superan lo establecido por la normativa Venezolana (0 NMP/100mL). Estadísticamente, no se evidenciaron diferencias significativas entre las piscinas para CT y CF antes y después de la limpieza. Las bacterias Gramnegativas predominaron, en E (84,21%) y B (71,92%), Klebsiella pneumoniae, Escherichia coli y Pseudomonas aeruginosa. A presentó mayor número de aislados Grampositivos (44,44%), identificándose Staphylococcus epidermidis y Enterococcus faecalis. Estos resultados indican una constante contaminación bacteriana y riesgo sanitario.

To evaluate the bacteriological quality of water pools in the city of Cumaná, Sucre state, Venezuela, water samples were collected in 1 public and 4 private pools, coded A to E, respectively, 2 samples were taken weekly for 2 months before and after a pool cleaning process. We determined pH, temperature and residual chlorine, aerobic mesophilic for total plate count, the Most Probable Number (MPN) for total coliforms (TC) and fecal (FC) and bacterial identification by conventional methods. The pH in the samples ranged between 6.8 and 7.3, temperature of 29 to 31ºC and chlorine residual of 0.3 to 0.5 mg/L The highest count of aerobic mesophilic bacteria was obtained in B with 6x10 2 . CFU/mL. In relation to the MPN, before cleaning, the CT highest value was obtained in D with 2.8 x10 3 and CT/100mL, E showed the highest CF value with 6x10 2 CF/100mL. After cleaning, the results for B indicated the CT highest values, 9.3 x10 2 CT/100mL, and in D and E the results had the CF highest values, 3x10 2 CF/100 mL. CF values in the samples before and after cleaning exceed the standards established by Venezuela (0 NMP/100mL). Statistically, no significant differences were found between pools for CT and CF before and after cleaning. Gram-negative bacteria predominated, being more frequent in E (84.21%) and B (71.92%), mainly, Klebsiella pneumoniae, Escherichia coli and Pseudomonas aeruginosa. Sample A had more isolated Gram-positive (44.44%), identified as Staphylococcus epidermidis and Enterococcus faecalis. These results indicate a constant bacterial contamination and health risk.

Prevalence and quantification of Listeria monocytogenes in beef offal at retail level in Selangor, Malaysia.

A total of 63 beef offal samples (beef liver = 16; beef lung = 14; beef intestine = 9; beef tripe = 15; beef spleen = 9) from three wet markets (A, B, and C) in Selangor, Malaysia were examined for the prevalence and microbial load of Listeria monocytogenes. A combination of the most probable number and polymerase chain reaction (MPN-PCR) method was employed in this study. It was found that L. monocytogenes detected in 33.33% of the beef offal samples. The prevalence of L. monocytogenes in beef offal purchased from wet markets A, B, and C were 22.73%, 37.50% and 41.18% respectively. The density of L. monocytogenes in all the samples ranged from < 3 up to > 2,400 MPN/g. The findings in this study indicate that beef offal can be a potential vehicle of foodborne listeriosis.

Prevalence and quantification of Listeria monocytogenes in beef offal at retail level in Selangor, Malaysia

A total of 63 beef offal samples (beef liver = 16; beef lung = 14; beef intestine = 9; beef tripe = 15; beef spleen = 9) from three wet markets (A, B, and C) in Selangor, Malaysia were examined for the prevalence and microbial load of Listeria monocytogenes. A combination of the most probable number and polymerase chain reaction (MPN-PCR) method was employed in this study. It was found that L. monocytogenes detected in 33.33% of the beef offal samples. The prevalence of L. monocytogenes in beef offal purchased from wet markets A, B, and C were 22.73%, 37.50% and 41.18% respectively. The density of L. monocytogenes in all the samples ranged from 3upto> 2,400 MPN/g. The findings in this study indicate that beef offal can be a potential vehicle of foodborne listeriosis.

Prevalence and quantification of Listeria monocytogenes in beef offal at retail level in Selangor, Malaysia

A total of 63 beef offal samples (beef liver = 16; beef lung = 14; beef intestine = 9; beef tripe = 15; beef spleen = 9) from three wet markets (A, B, and C) in Selangor, Malaysia were examined for the prevalence and microbial load of Listeria monocytogenes. A combination of the most probable number and polymerase chain reaction (MPN-PCR) method was employed in this study. It was found that L. monocytogenes detected in 33.33% of the beef offal samples. The prevalence of L. monocytogenes in beef offal purchased from wet markets A, B, and C were 22.73%, 37.50% and 41.18% respectively. The density of L. monocytogenes in all the samples ranged from 3upto> 2,400 MPN/g. The findings in this study indicate that beef offal can be a potential vehicle of foodborne listeriosis.

Evaluation of microorganisms with sulfidogenic metabolic potential under anaerobic conditions

The aim of this work was to identify groups of microorganisms that are capable of degrading organic matter utilizing sulfate as an electron acceptor. The assay applied for this purpose consisted of running batch reactors and monitoring lactate consumption, sulfate reduction and sulfide production. A portion of the lactate added to the batch reactors was consumed, and the remainder was converted into acetic, propionic and butyric acid after 111 hours of operation These results indicate the presence of sulfate-reducing bacteria (SRB) catalyzing both complete and incomplete oxidation of organic substrates. The sulfate removal efficiency was 49.5% after 1335 hours of operation under an initial sulfate concentration of 1123 mg/L. The SRB concentrations determined by the most probable number (MPN) method were 9.0x10(7) cells/mL at the beginning of the assay and 8.0x10(5) cells/mL after 738 hours of operation.

Standardization of a quantification method for Salmonella spp. and Shigella spp. in specific liquid media / Estandarización de un método de recuento para Salmonella spp. y Shigella spp. en medios de cultivo líquidos especializados

Introduction: Chlorination is the most widely used disinfection process for drinking water production. The formation of chlorination carcinogenic by-products and chlorine intoxication by direct manipulation in small communities has motivated the study of alternative disinfection processes. In this sense, processes of advanced oxidation (PAOs) have yielded promising results. Escherichia coli (E. coli) is customarily used as faecal bacterial indicator to determine the efficiency of disinfection processes. However, it has been shown that E. coli is less resistant to disinfection than other enteric bacteria such as Shigella  spp. and Salmonella  spp. Additionally, the viable non-culturable (VNC) state yields bacteria which are not detectable on many culture media. Objective: The main objective is to standardize a method for counting Salmonella  spp. and Shigella  spp. in specific liquid media to reliably quantify the bacteriological potential risk related to disinfection processes based on PAO. Methods: The study followed a randomized bi-factorial experimental design and the Duncan multiple comparison test. This design allowed the selection of specific liquid media to fittingly standardize the counting of Salmonella  spp. and Shigella  spp. Results: We found that the best broth for counting Salmonella typhimurium strain at different concentrations in pure and mixed cultures was the Rappaport broth RP, the EE broth also allowed growing the two bacterial species tested in this research. Nonetheless, the latter results suggest the use of additional tests for this particular broth. Discussion: There was a variation in the counting results when pure cultures were used compared to those obtained from mixtures of microorganisms. It was also noted that Salmonella typhimurium and Shigella sonnei, were recovered from minimal concentrations in both RP and EE broths, respectively. To some extent, this suggests an additional confirmative method when using the EE® broth...

Introducción: La cloración es el método más usado para desinfectar aguas de consumo. La formación de subproductos cancerígenos y las intoxicaciones por manipulación directa en pequeñas comunidades, han motivado el estudio de procesos alternativos. Los procesos de oxidación avanzada (PAOS), han arrojado resultados prometedores, utilizando el indicador bacteriano Escherichia coli (E. coli), con el método recuento en placa. Sin embargo, también se ha demostrado que E. coli es menos resistente a la desinfección que otras bacterias entéricas como Shigella y Salmonella y que estos procesos generan bacterias viables que no se cultivan durante el proceso, y no se descubren en medios sólidos. Objetivo: Estandarizar un método de recuento de Salmonella sp. y Shigella sp., en medios de cultivo líquidos especializados, que permita valorar de forma confiable el riesgo bacteriológico en procesos de desinfección PAOS. Métodos: En el presente trabajo se ensayaron y seleccionaron medios líquidos especializados, con los que se estandarizó el recuento de Salmonella sp. y Shigella sp., mediante un diseño experimental aleatorizado bifactorial y la prueba de comparaciones múltiples de Duncan. Resultados: Se encontró que el mejor caldo para recuperar a S. typhimurium a diferentes concentraciones, en cultivos puros y mezclas, fue el caldo Rappaport de Merck (RP). El caldo de enriquecimiento para entero bacterias de Oxoid (EE), permitió un buen crecimiento de las dos especies objeto de esta investigación. Lo cual sugiere el empleo de pruebas adicionales cuando se use caldo EE para NMP. Discusión: Se observó una variación en el recuento cuando se usaron cultivos puros, comparado con la obtenida a partir de mezclas de microorganismos. Sin embargo, S. typhimurium. y Shigella sonnei logran ser recuperadas de concentraciones mínimas en los caldos RP, respectivamente...