Length and caliber of the rectosigmoid colon among patients with Chagas disease and controls from areas at different altitudes

Introduction In this study, we investigated radiological changes in the sigmoid colon in chagasic patients by comparing their colon lengths and caliber with those of non-chagasic living in the same region and non-chagasic living at high altitudes. Methods A total of 317 individuals were evaluated using clinical, serological and radiological methods and divided into three groups: 1) one hundred and nine non-chagasic individuals from Uberaba, Brazil; 2) sixty-one non-chagasic from Puno, Peru; 3) one hundred forty-seven chagasics examined in Uberaba, being 62 without megacolon (3A), 72 with megacolon (3B) and 13 with doubtful diagnosis of megacolon (3C). Results In group 2, the sigmoid colon had a significantly larger caliber (p=0.001) and the rectosigmoid colon was longer (p<0.001) than group 1. In subgroup 3A, the sigmoid colon (p<0.001) and rectum (p<0.001) had a significantly larger caliber and the rectosigmoid was longer (p<0.001) than that of the non-chagasic individuals. In subgroup 3B, the rectosigmoid was longer in all patients, and the caliber of the sigmoid was significantly larger than that of subjects in subgroups 3A and 3C (p<0.001). Conclusions Morphometric analysis confirms that Chagas disease may increase the caliber and length of the rectosigmoid. Our results suggest that altitude, ethnicity and diet may have influenced the size and length of the rectosigmoid of andean patients.

Biology of Triatoma carcavalloi Jurberg, Rocha & Lent, 1998 under laboratory conditions

Introduction Triatoma carcavalloi is a wild species that is found in sympatry with Triatoma rubrovaria and Triatoma circummaculata, which are vectors of Trypanosoma cruzi currently found in rural areas of Rio Grande do Sul, Brazil. Methods Fertility was assessed and to determine the incubation period, the eggs were observed until hatching. The first meal was offered to 1st stage nymphs. The intermolt period was also determined. The number of blood meals was quantified at each nymphal stage and the resistance to fasting as the period between ecdysis and death. Mortality was assessed and longevity was determined by recording the time that elapsed from molting to the adult stage and until death. The developmental cycle was assessed by recording the length in days of each stage from molting to adult hood. Results The average incubation period was 22.7 days. The average first meal occurred 3.1 days after hatching. The 5th stage nymph to adult intermolting period was the longest at 193.4 days. The average number of feedings during nymphal development was 13.4. The resistance to fasting assay indicated that the 3rd, 4th and 5th stage nymphs presented higher resistance than did adults. The highest mortality rate was observed in the 3rd stage nymphs (22.2%). The average length of adult survival was 25.6 weeks, and the average total life cycle lasted 503.4 days. Conclusions This study is the first report on the biology of T. carcavalloi that fed on mice. The presented findings expand the bionomic knowledge of these species.

Practice variations in the production of health

This study investigates three questions related to medical practice variation. First, it tests whether average length of stay across Portuguese National Health Service hospitals varies when controlling for differences in patients’ characteristics. Second, it looks at hospital-level characteristics in order to find out whether these are able to explain differences in average length of stay across hospitals. Finally, it proposes a best practice average length of stay for each of the six episodes of care analyzed. To perform the analysis, administrative data from the Diagnosis-Related groups’ data set for the year of 2012 was used. A replication of a hierarchical two-stage model with hospital fixed effects was carried out. The results show that after taking patients’ characteristics into account, variation in average length of stay across hospitals exists. This variation cannot be explained by hospital-level characteristics.

Analysis of outcome and cancer stem cells status in patients with resectable pancreatic adenocarcinoma

RESUMO: Actualmente, a única possibilidade de cura para doentes com adenocarcinoma do pâncreas (PDAC) é a ressecção cirúrgica, no início deste estudo, perguntamo-nos se os predictores clínico-patológicos clássicos de prognostico poderiam ser validados em uma grande cohort de doentes com cancro do pâncreas ressecável e se outros predictores clínicos poderiam ter um papel na decisão de que doentes beneficiariam de ressecção cirúrgica. No capítulo 2, observamos que até 30% dos doentes morrem no primeiro ano após a ressecção cirúrgica, pelo que o nosso objectivo foi determinar factores pré-operatórios que se correlacionam com mortalidade precoce após ressecação cirúrgica com recurso a um instrumento estatisticamente validado, o Charlson-Age Comorbidity Index (CACI), determinamos que um CACI score superior a 4 foi preditivo de internamentos prolongados (p <0,001), complicações pós-operatórias (p = 0,042), e mortalidade em 1 ano pós- ressecção cirúrgica (p <0,001). Um CACI superior a 6 triplicou a mortalidade no primeiro ano pós-cirurgia e estes doentes têm menos de 50% de probabilidade de estarem vivos um ano após a cirurgia. No capítulo 3, o nosso objectivo foi identificar uma proteína de superfície que se correlacionasse estatisticamente com o prognostico de doentes com adenocarcinoma do pâncreas e permitisse a distinção de subgrupos de doentes de acordo com as suas diferenças moleculares, perguntamo-nos ainda se essa proteína poderia ser um marcador de células-estaminais. No nosso trabalho anterior observamos que as células tumorais na circulação sanguínea apresentavam genes com características bifenotípica epitelial e mesenquimal, enriquecimento para genes de células estaminais (ALDH1A1 / ALDH1A2 e KLF4), e uma super-expressão de genes da matriz extracelular (colagénios, SPARC, e DCN) normalmente identificados no estroma de PDAC. Após a avaliação dos tumores primários com RNA-ISH, muitos dos genes identificados, foram encontrados co-localizando em uma sub-população de células na região basal dos ductos pancreáticos malignos. Além disso, observamos que estas células expressam o marcador SV2A neuroendócrino, e o marcador de células estaminais ALDH1A1/2. Em comparação com tumores negativos para SV2, os doentes com tumores SV2 positivos apresentaram níveis mais baixos de CA 19-9 (69% vs. 52%, p = 0,012), tumores maiores (> 4 cm, 23% vs. 10%, p = 0,0430), menor invasão de gânglios linfáticos (69% vs. 86%, p = 0,005) e tumores mais diferenciados (69% vs. 57%, p = 0,047). A presença de SV2A foi associada com uma sobrevida livre de doença mais longa (HR: 0,49 p = 0,009) bem como melhor sobrevida global (HR: 0,54 p = 0,018). Em conjunto, esta informação aponta para dois subtipos diferentes de adenocarcinoma do pâncreas, e estes subtipos co-relacionam estatisticamente com o prognostico de doentes, sendo este subgrupo definido pela presença do clone celular SV2A / ALDH1A1/2 positivo com características neuroendócrinas. No Capítulo 4, a expressão de SV2A no cancro do pâncreas foi validado em linhas celulares primárias. Demonstramos a heterogeneidade do adenocarcinoma do pâncreas de acordo com características clonais neuroendócrinas. Ao comparar as linhas celulares expressando SV2 com linhas celulares negativas, verificamos que as linhas celulares SV2+ eram mais diferenciadas, diferindo de linhas celulares SV2 negativas no que respeita a mutação KRAS, proliferação e a resposta à quimioterapia. No capítulo 5, perguntamo-nos se o clone celular SV2 positivo poderia explicar a resistência a quimioterapia observada em doentes. Observamos um aumento absoluto de clones celulares expressando SV2A, em múltiplas linhas de evidência - doentes, linhas de células primárias e xenotransplantes. Embora, tenhamos sido capazes de demonstrar que o adenocarcinoma do pâncreas é uma doença heterogénea, consideramos que a caracterização genética destes clones celulares expressando SV2A é de elevada importância. Pretendemos colmatar esta limitação com as seguintes estratégias: Após o tratamento com quimioterapia neoadjuvante na nossa coorte, realizamos microdissecação a laser das amostras primarias em parafina, de forma a analisar mutações genéticas observadas no adenocarcinoma pancreático; em segundo lugar, pretendemos determinar consequências de knockdown da expressão de SV2A em nossas linhas celulares seguindo-se o tratamento com gemicitabina para determinação do papel funcional de SV2A; finalmente, uma vez que os nossos esforços anteriores com um promotor - repórter e SmartFlare ™ falharam, o próximo passo será realizar RNA-ISH PrimeFlow™ seguido de FACS e RNA-seq para caracterização deste clone celular. Em conjunto, conseguimos provar com várias linhas de evidência, que o adenocarcinoma pancreático é uma doença heterogénea, definido por um clone de células que expressam SV2A, com características neuroendócrinas. A presença deste clone no tecido de doentes correlaciona-se estatisticamente com o prognostico da doença, incluindo sobrevida livre de doença e sobrevida global. Juntamente com padrões de proliferação e co-expressão de ALDH1A1/2, este clone parece apresentar um comportamento de células estaminais e está associado a resistência a quimioterapia, uma vez que a sua expressão aumenta após agressão química, quer em doentes, quer em linhas de células primárias.----------------------------- ABSTRACT: Currently, the only chance of cure for patients with pancreatic adenocarcinoma is surgical resection, at the beginning of my thesis studies, we asked if the classical clinicopathologic predictors of outcome could be validated in a large cohort of patients with early stage pancreatic cancer and if other clinical predictors could have a role on deciding which patients would benefit from surgery. In chapter 2, we found that up to 30% of patients die within the first year after curative intent surgery for pancreatic adenocarcinoma. We aimed at determining pre-operative factors that would correlate with early mortality following resection for pancreatic cancer using a statistically validated tool, the Charlson-Age Comorbidity Index (CACI). We found that a CACI score greater than 4 was predictive of increased length of stay (p<0.001), post-operative complications (p=0.042), and mortality within 1-year of pancreatic resection (p<0.001). A CACI score of 6 or greater increased 3-fold the odds of death within the first year. Patients with a high CACI score have less than 50% likelihood of being alive 1 year after surgery. In chapter 3 we aimed at identifying a surface protein that correlates with patient’s outcome and distinguishes sub-groups of patients according to their molecular differences and if this protein could be a cancer stem cell marker. The most abundant class of circulating tumor cells identified in our previous work was found to have biphenotypic features of epithelial to mesenchymal transition, enrichment for stem-cell associated genes (ALDH1A1/ALDH1A2 and KLF4), and an overexpression of extracellular matrix genes (Collagens, SPARC, and DCN) normally found in the stromal microenvironment of PDAC primary tumors. Upon evaluation of matched primary tumors with RNA-ISH, many of the genes identified were found to co-localize in a sub-population of cells at the basal region of malignant pancreatic ducts. In addition, these cells expressed the neuroendocrine marker SV2A, and the stem cell marker ALDH1A1/2. Compared to SV2 negative tumors, patients with SV2 positive tumors were more likely to present with lower CA 19-9 (69% vs. 52%, p = 0.012), bigger tumors (size > 4 cm, 23% vs. 10%, p= 0.0430), less nodal involvement (69% vs. 86%, p = 0.005) and lower histologic grade (69% vs. 57%, p = 0.047). The presence of SV2A expressing cells was associated with an improved disease free survival (HR: 0.49 p=0.009) and overall survival (HR: 0.54 p=0.018) and correlated linearly with ALDH1A2. Together, this information points to two different sub-types of pancreatic adenocarcinoma, and these sub-types correlated with patients’ outcome and were defined by the presence of a SV2A/ ALDH1A1/2 expressing clone with neuroendocrine features. In Chapter 4, SV2A expression in cancer was validated in primary cell lines. We were able to demonstrate pancreatic adenocarcinoma heterogeneity according to neuroendocrine clonal features. When comparing SV2 expressing cell lines with SV2 negative cell lines, we found that SV2+ cell lines were more differentiated and differ from SV2 negative cell lines regarding KRAS mutation, proliferation and response to chemotherapy. In Chapter 5 we aimed at determining if this SV2 positive clone could explain chemoresistance observed in patients. We found an absolute increase in SV2A expressing cells, with multiple lines of evidence, in patients, primary cell lines and xenografts. Although, we have been able to show evidence that pancreatic adenocarcinoma is a heterogeneous disease, our findings warrant further investigation. To further characterize SV2A expressing clones after treatment with neoadjuvant chemotherapy in our cohort, we have performed laser capture microdissection of the paraffin embedded tissue in this study and will analyze the tissue for known genetic mutations in pancreatic adenocarcinoma; secondly, we want to know what will happen after knocking down SV2A expression in our cell lines followed by treatment with gemcitabine to determine if SV2A is functionally important; finally, since our previous efforts with a promoter – reporter and SmartFlare™ have failed, we will utilize a novel PrimeFlow™ RNA-ISH assay followed by FACS and RNA sequencing to further characterize this cellular clone. Overall our data proves, with multiple lines of evidence, that pancreatic adenocarcinoma is a heterogeneous disease, defined by a clone of SV2A expressing cells, with neuroendocrine features. The presence of this clone in patients’ tissue correlates with patient’s disease free survival and overall survival. Together with patterns of proliferation and ALDH1A1/2 co-expression, this clone seems to present a stem-cell-like behavior and is associated with chemoresistance, since it increases after chemotherapy, both in patients and primary cell lines.

The Larval Development of Palaemonid Shrimps from the Amazon region reared In the laboratory. VII. Abbreviated development of Pseudopalaemon amazonensis Ramos-Porto, 1979 (Crustacea: Decapoda: Caridea)

Larval development of the freshwater shrimp Pseudopalaemon amazonensis Ramos-Porto was studied in the laboratory based on the offspring of ovigerous females collected in a small “terra-firme” forest stream near Manaus, Brazil. Ovigerous females with a mean total length of 36.5 ± 1.9 mm carried 13-19 eliptical, yolk-rich eggs measuring 2.55 ± 0.16 x 1.64 ± 0.11 mm. The larval period consisted of 3 benthie stages and the larvae accomplished metamorphosis after 7-8 days without feeding. The newly-hatched larva had sessile eyes and all appendages, except for the uropods; chelipeds were present as uniramous buds, but walking legs were fully developed and functional. Descriptions and illustrations of the 3 larval and first juvenile stages are presented.

Bioecology of the crab Ucides cordatus (Crustacea, Decapoda) in mangroves influenced by the Amazon River, Brazil

The mangrove crab (Ucides cordatus) is a valuable fishery resource, overfished along the Brazilian coast. This study aimed to obtain bioecological data on this crab along the coast of the State of Amapá. Six bimonthly samplings were conducted between December 2008 and January 2010. Transects were used to estimate the density (burrows m-2) and population abundance (individuals m-2). All the animals were subjected to biometrics, with females being classified according to their stage of gonadal maturation. The mean density (1.09 burrows m-2) and abundance (0.31 individuals m-2) were influenced by the climate with the highest values in summer (1.17 burrows m-2 and 0.34 individuals m-2). The male to female ratio was 1.38:1 showing significant difference in the proportion of sexes. The individuals showed sexual dimorphism, with linear measurements significantly higher in males. The sampled animals also had larger carapace length and width (CL and CW) compared to crabs studied in other Brazilian states. There was a positive relationship between CW and CL and individual weight (IW) and CW for males (R² = 0.83 and 0.90) and females (R² = 0.79 and 0.84). The growth was negative allometric (CL increases to a lesser extent than CW) for both sexes. The highest frequency of ovigerous females (78%) and in maturation stage IV (38%) occurred in the CW size class between 59.8 and 67.5 mm. The peak of mature females occurred in May and August, showing a reproductive period different from those in other Brazilian states.

Ichthyometry and electrical bioimpedance analysis to estimate the body composition of tambatinga

Body composition analysis is relevant to characterize the nutritional requirements and finishing phase of fish. The aim of this study was to investigate the relationship between ichthyometric (weight, total and standard length, density and yields), bromatological (fat, protein, ash and water content) and bioelectrical-impedance-analysis (BIA) (resistance, reactance, phase angle and composition indexes) variables in the hybrid tambatinga (Colossoma macropomum × Piaractus brachypomus). In a non-fertilized vivarium, 520 juveniles were housed and fed commercial rations. Then, 136 days after hatching (DAH), 15 fish with an average weight of 37.69 g and average total length of 12.96 cm were randomly chosen, anesthetized (eugenol) and subjected to the first of fourteen fortnightly assessments (BIA and biometry). After euthanasia, the following parts were weighed: whole carcass with the head, fillet, and skin (WC); fillet with skin (FS); and the remainder of the carcass with the head (CH). Together, FS and CH were ground and homogenized for the bromatological analyses. Estimates of the body composition and yields of tambatinga, with models including ichthyometric and BIA variables, showed correlation coefficients ranging from 0.81 (for the FS yield) to 1,00 (for the total ash). Similarly, models that included only BIA variables had correlation coefficients ranging from 0.81 (FS and CH yields) to 0.98 (for the total ash). Therefore, in tambatinga, the BIA technique allows the estimation of the yield of the fillet with skin and the body composition (water content, fat, ash, and protein). The best models combine ichthyometric and BIA variables.

Reproductive biology of Macrobrachium surinamicum (Decapoda: Palaemonidae) in the Amazon River mouth

Macrobrachium surinamicum is an indigenous prawn distributed from the lower Amazon and Tocantins river basins to Venezuela in the Orinoco Delta region. It is common bycatch fauna of Macrobrachium amazonicum artisan fishing in the states of Pará and Amapá. The aim of this study was to investigate aspects on reproductive biology (reproductive period, size of sexual maturity population, fecundity, reproductive output and recruitment) of M. surinamicum from four important areas to artisanal prawn fishing located at the Amazon River mouth (Amapá and Pará). The specimens were captured using 20 handcrafted traps called "matapi". A number of 675 prawns were captured, 258 males, 409 females and eight juveniles, resulting in 1:1.6 (Male: Female) sex ratio. The reproductive peak period occurred from March to July, coinciding with the higher rainfall period. The juvenile prawn occurred only in May and July. Total length of egg-bearing females ranged from 12.12 to 38.30 mm, with mean female length at first maturity (L50) of 23.7 mm. Fecundity increased with prawn size and varied between 174 and 1780 eggs per female. Mean egg volume increased gradually from 0.031 (Stage I) to 0.060 mm³ (Stage III) during embryogenesis. Macrobrachium surinamicum depends on brackish water to complete the larval development. Irrespective of female size, reproductive output of M. surinamicum varied between 4.3 % and 35.5 % of their body weight for egg production. The knowledge of the reproductive biology reported in the present study is an important tool to define strategies to preserve M. surinamicum in Amazon River mouth.

Resource partitioning and ecomorphological variation in two syntopic species of Lebiasinidae (Characiformes) in an Amazonian stream

ABSTRACTResource partitioning is important for species coexistence. Species with similar ecomorphology are potential competitors, especially when phylogenetically close, due to niche conservatism. The aim of this study was to investigate the resource partitioning among populations of two species of lebiasinids (Copella nigrofasciata and Pyrrhulina aff. brevis) that co-occur in a first-order Amazonian stream, analyzing the trophic ecology, feeding strategies and ecomorphological attributes related to the use of food and space by these species. Fish were captured in May and September 2010. The stomach contents of 60 individuals were analyzed and quantified volumetrically to characterize the feeding ecology of both species. Eleven morphological attributes were measured in 20 specimens and combined in nine ecomorphological indices. Both species had an omnivorous-invertivorous diet and consumed predominantly allochthonous items. Both showed a tendency to a generalist diet, but intrapopulational variation in resource use was also detected. Overall feeding niche overlap was high, but differed between seasons: low during the rainy season and high in the dry season. In the latter, the food niche overlap was asymmetric because C. nigrofasciata consumed several prey of P. aff. brevis, which reduced its food spectrum. The ecomorphological analysis suggests that C. nigrofasciatahas greater swimming capacity (greater relative length of caudal peduncle) than P. aff. brevis, which has greater maneuverability and tendency to inhabit lentic environments (greater relative depth of the body). Our results demonstrate that these species have similar trophic ecology and suggest a spatial segregation, given by morphological differences related to locomotion and occupation of habitat, favoring their coexistence.

Age, growth and reproductive biology of whiting Merlangius merlangus (Linnaeus 1758) in the Celtic Sea

Age, growth and reproductive biology were investigated for whiting (Merlangius merlangus) captured from the Celtic Sea (ICES division Vllg), for the period January 2001 to January 2002. Females dominated the sex ratio of 1: 2.25. The relative abundance of females exceeded the number of males in all length classes. The relationship between weight (g) and total length (cm) was the same for male and female whiting. A total of 973 fish were aged and the maximum age recorded was 11 years. Results from an intercalibration exercise showed 87% agreement in age readings between the author and an expert in ageing whiting at the Marine Institute. Females were dominated by 2 year olds, while males were dominated by 3 year olds. Lx was estimated as 38cm and a growth rate was calculated K = 0.3769 year'1. Females were fully recruited to the fishery at 3 years of age, while the age at full recruitment (tr) for males was 4 years. Female whiting spawned from late February to June 2001 and matured at a total length of 23 cm in their first year. Female whiting reached L5o at a total length of 28 cm and 2.7 years of age. Male whiting spawned from February to June 2001. They matured at a total length of 21 cm and in their first year. Male whiting reached L50 at a total length of 30.4 cm and 3.6 years of age. The following critical points should be taken into account in the management of the Celtic Sea whiting stock: An Fpa should be established in order to assess the current level of fishing mortality; The maturity ogives need further study; The extent of gutting of large fish before landing by fishers in the fleet should be investigated and the apparent decline in size of 4 - 7 year old fish in the Celtic Sea between 1996 and 2001 needs to be assessed.

Age, growth and reproductive biology of Plaice, (Pleuronectes platessa L.) in Irish waters, 2003-2005

The current study presents data on age and growth for plaice (Pleuronectes platessa L.) sampled between November 2003 and February 2005 in ICES areas Via (northwest coast of Ireland), Vila (Irish Sea), Vllg (Celtic Sea), VDj (southwest coast of Ireland) and VHb (west coast of Ireland), and data on the reproductive biology and maturity of plaice in ICES area Vllb (west coast of Ireland). This is the first detailed account of the biology of plaice for some of these areas. It is intended that this study will improve understanding of the life cycle of plaice and help fisheries scientists to better predict the effect of fishing effort on Irish plaice stocks. The overall length range found for plaice was 9-51.99cm TL, with a length range of 9-5 lcm TL for females and 9-40cm for males. In all ICES areas the length range for female fish was larger than for male fish. The age range of plaice sampled during this study was 1 to 16 years. In all ICES areas females had a greater range in ages and fish in the larger age groups. From analysis of length and age data it was concluded that there was a significant difference (P=0.000) in growth rate of males and females between ICES areas sampled in March 2004. The highest rate of fishing mortality was determined for ICES area Via (F=1.06) and the lowest for ICES area Vila (F=0.56). In each ICES area male and female plaice have fully recruited to the population by age 4, with the exception of females in ICES area Via, for which a tr value of 5 years was determined. Length at first maturity (L50%) was determined to be 23cm and 21cm for males and females respectively. Age at first maturity (A50%) was determined to be 3 years for both males and females. It was found that males and females in ICES areas Vllb, Vila and Via are well above the length and age at first maturity when they are recruited to the fishery. In ICES area Vllb female plaice spawn from November to March, with peak spawning occurring in February, and male plaice spawn from November to April, with peak spawning occurring in November. Spawning females had an age range of 2 to 10 years and spawning males had an age range of 2 to 7 years. From the oocyte length frequency distributions, it was determined that the plaice is a determinate batch spawner. During this investigation a total of 177 ovaries and 127 testes were staged using both macroscopic and histological criteria. The overall percentage of maturity stages which compared favorably between the two assessment methods was 22.03% for female plaice and 37.80% for male plaice. In general, the findings of this study indicate that there was a very poor match between the macroscopic and histological assessment methods. Given that the histological determination of these stages is based on the observation of a distinct set of developmental features, it is expected that it would be more accurate to use histologically assessed gonads to calculate the annual percentage maturity assessment. The biology of plaice in the areas studied is compared with previous studies of plaice in Irish and European waters.

The relation of the concentration of macronutrients in the substrate and in the foliage to cell wall thickness and cellulose concentration in the xylem of slash pine (Pinus elliotti)

Sand culture experiments, using a sub-irrigation technique, were installed in order to find out the effects of the macronutrients N, P, K, Ca, Mg and S on growth, aspect, mineral composition, length of fibers, thickness of cell wall and cellulose concentration in slash pine. The aim was to obtain, under controlled conditions, basic information which could eventually lead to practical means designed to increase the rate of growth and to make of slash pine a richer source of cellulose. Nitrogen, Phosphorus, Potassium Experiment A 3 x 3 x 3 factorial design with two replicates was used. Nitrogen was supplied initially at the levels of 25, 50 and 100 ppm; phosphorus was given at the rates of 5, 10 and 20 ppm; potassium was supplied at the rates of 25, 50 and 100 ppm; six months after the experiment was started the first level for each element was dropped to zero. Others macro and all micronutrients were supplied at uniform rates. Fifteen hours of illumination per day were provided. The experimental technique for growing the slash pine seedlings proved quite satisfactory. Symptoms of deficiency of nitrogen, phosphorus and potassium were observed, described and recorded in photographs and water colors. These informations will help to identify abnormalities which may appear under field conditions. Chemical analysis of the several plant parts, on the other hand, give a valuable means to assess the nutritional status of slash pine, thus confirming when needed, the visual diagnosis. The correctness of manurial pratices, on the other hand, can be judged with the help of the analytical data tabulated. Under the experimental conditions nitrogen caused the highest increases on growth, as measured by increments in height and dry weights, whereas the effects of phosphorus and potassium were less marked. Cellulose concentration was not significantly affected by the treatments used. Higher levels of N seemed to decrease both length of fiber elements and the thickness of cell wall. The effects of P and K were not well defined. Calcium, Magnesium, Sulfur Experiment A 3 x 3 x 3 factorial design with two replicates was used. Calcium was supplied initially at the levels of 12.5, 25 and 50 ppm; magnesium and sulfur were given at the rates of 6, 12.5 and 25 ppm. Other macro and micronutrients were supplied at uniform rates, common to all treatments. Three months after starting the experiment the first level for each element was dropped to zero. Symptoms of deficiency of calcium, magnesium and sulfur were observed, described and recorded as in the case of the previous experiment. Chemical analysis were made, both for mineral content and cellulose concentration. Length of fibers and thickness of cell wall were measured. Both calcium and magnesium increase height, sulfur failing to give significant response. Dry weight was beneficially affected by calcium and sulfur. The levels of calcium, magnesium and sulfur in the needles associated with deficiency and maximum growth are comparable with those found in the literature. Cellulose concentration increased when the level of sulfur in the substrate was raised. The thickness of cell wall was negatively affected by the treatments; no effect was observed with regards to length of fibers.

Biologia reprodutiva de Apidoras fuscoguttatus (Siluriformes, Callichthyidae) em riacho de cabeceira da bacia do alto Rio Paraná

The reproductive biology of Aspidoras fuscoguttatus Nijssen & Isbrücker, 1976 from a stream in São José do Rio Preto, northwestern São Paulo State, Brazil, was monthly investigated in the period of August 1999 to July 2000. Measurements of total length, body weight, gonadal weight and macroscopic assessment of gonadal maturation were performed. Environmental parameters were considered in order to verify associations with the reproductive period. Populational structure showed total length amplitude between 14.2 and 50.8 mm. Pronounced sexual dimorphism was verified. The largest mean values of gonadosomatic relation for females coincided with the rainy season (November to March). Mean length at first sexual maturity was different for males (30.5 mm) and females (37.1 mm). Fecundity varied between 51 and 166 oocytes. Gonadal maturation curve, frequency of maturation stages and size frequency distributions of oocytes in mature ovaries revealed a long reproductive period, suggesting fractional spawning.

Dinâmica reprodutiva de Oligosarcus jenynsii e O. robustus (Characiformes, Characidae) na lagoa Fortaleza, Rio Grande do Sul, Brasil

Oligosarcus jenynsii (Günther, 1864) and O. robustus Menezes, 1969 are fish species distributed in Rio Grande do Sul, Brazil, Uruguay, and northern Argentina. The reproductive period and recruitment, sex ratio, absolute and relative fecundity, and body length at first gonadal maturation of the two carnivorous species from Fortaleza Lagoon were analized. The specimens were sampled monthly, from May 2000 to April 2001, with fishing effort of 24 hours/month, using stationary gillnets of different mesh sizes and seine net (three samples per edge). The records of each individual included total length, total weight, gonad weight, sex and gonadal maturity stage. The reproductive period of both O. jenynsii and O. robustus ranges from May/June to November/December, according to the bimonthly variation of the mean values of gonosomatic index, and the relative frequencies of the gonadal maturation stages. Recruitment of new individuals to the population occurs from November/December to March/April. The sex ratio is different from 1:1 for O. jenynsii and similar to 1:1 for O. robustus. The mean absolute fecundity, calculated by counting sub-sampled oocytes from mature females, was 14,483 oocytes for O. jenynsii, and 16,308 oocytes for O. robustus. The first maturation curve shows that O. jenynsii begins to reproduce between 84 mm and 104 mm (total length), and O. robustus between 126 mm and 146 mm, probably at similar ages.

Morphometric analysis of Capitella capitata (Polychaeta, Capitellidae)

Accurate size measurements are fundamental in characterizing the population structure and secondary production of a species. The purpose of this study was to determine the best morphometric parameter to estimate the size of individuals of Capitella capitata (Fabricius, 1780). The morphometric analysis was applied to individuals collected in the intertidal zones of two beaches on the northern coast of the state of São Paulo, Brazil: São Francisco and Araçá. The following measurements were taken: the width and length (height) of the 4th, 5th and 7th setigers, and the length of the thoracic region (first nine setigers). The area and volume of these setigers were calculated and a linear regression analysis was applied to the data. The data were log-transformed to fit the allometric equation y = ax b into a straight line (log y = log a + b * log x). The measurements which best correlated with the thoracic length in individuals from both beaches were the length of setiger 5 (r² = 0.722; p<0.05 in São Francisco and r² = 0.795; p<0.05 in Araçá) and the area of setiger 7 (r² = 0.705; p<0.05 in São Francisco and r² = 0.634; p<0.05 in Araçá). According to these analyses, the length of setiger 5 and/or the area of setiger 7 are the best parameters to evaluate the growth of individuals of C. capitata.