Formalin-fixed paraffin-embedded tissues for microbiome analysis in rainbow trout (Oncorhynchus mykiss).

The gut microbiomes of rainbow trout (Oncorhynchus mykiss) reared at 16° and 22 °C were determined using formalin-fixed paraffin-embedded tissues (FFPE) and compared to fresh frozen tissue. The data revealed microbiomes could be successfully determined using FFPE tissue opening a new horizon in studying intestinal microbiota using archived histological samples.

Obtaining bioproducts by slow pyrolysis of coffee and cocoa husks as suitable candidates for being used as soil amendment and source of energy / Obtención de bioproductos por pirólisis lenta de cascarillas de café y cacao para su posible uso como fuentes de energía y fertilizantes / Obtenção de bioprodutos por pirólise lenta de cascas de café e cacau para possível uso como fontes de energia e fertilizantes

Abstract The agricultural economic policy of Cuba pretends to triplicate the crops of coffee and cocoa by 2021, a measure which will bring about both an increase in biomass waste and the need to find a proper method for its disposal. Slow pyrolysis process can transform lignocellulosic waste into added value products as biochar, bio-oil and gas. The present research evaluated the yield and the theoretical potential energy of co-products (bio-char, bio-oil, and gas) from coffee and cocoa seed husks using the pyrolysis process as a source of friendly and renewable energy. Results indicated that coffee husks are more suitable for the production of gas with a yield of 40.4%, while cocoa seed husks are better suited for the production of bio-oil, with a yield of 37.4%. For 2021 the theoretical energy potential estimated is 8291 MWh, equivalent to 716000 kg for the coffee husks, and 1384 MWh equivalent to 121000 kg for the cocoa seed husks. The bio-char characterization indicated high contents of carbon, calcium, potassium, nitrogen, and oxygen, which makes it a suitable candidate for being used as a soil amendment. The conversion of coffee and cocoa seed husks into added value products through slow pyrolysis process will help clean the environment, decrease the greenhouse effect, and will aid farmers in the rural populations by providing them with an additional source of income.

Resumen La política económica agrícola de Cuba pretende triplicar los cultivos de café y cacao para el año 2021, lo cual implica un aumento de residuos de biomasa y la necesidad de encontrar un uso adecuado para su eliminación. El proceso de pirólisis lenta permite convertir los residuos lignocelulósicos en productos de valor agregado como bio-carbón, bio-combustible y gas. En este trabajo se evaluó el rendimiento y el potencial energético teórico de estos co-productos como fuente de energía renovable. Las cascaras de café son más adecuadas para la producción de gas con un rendimiento del 40,4%, mientras que las de semilla de cacao para bio-combustible, con un rendimiento del 37,4%. Para el 2021, el potencial energético teórico estimado es de 8291 MWh, equivalente a 716000 kg para las cascaras de café; y de 1384 MWh equivalente a 121000 kg para las semillas de cacao. La caracterización del bio-carbón indicó altos contenidos de carbono, calcio, potasio, nitrógeno y oxígeno, lo que los convierte en un candidato adecuado para su uso como fertilizante del suelo. La conversión de estas biomasas en productos de valor agregado a través del proceso de pirólisis lenta contribuirá a mantener un ambiente limpio, reducir el efecto invernadero y ayudará a generar un ingreso adicional a los agricultores de las poblaciones rurales.

Resumo A política económica agrícola de Cuba visa triplicar as safras de café e cacau até 2021, o que significa um aumento nos resíduos de biomassa e a necessidade de encontrar usos adequados para sua eliminação. O processo de pirólise lenta permite converter lixo lignocelulósico em produtos de alto valor agregado, como bio-carvão, biocombustível e gás. No presente trabalho, avaliou-se o desempenho e o potencial energético teórico de co-produtos (bio-carvão, biocombustível e gás) do processo de pirólise lenta de casca de café e de cascas de sementes de cacau como fonte de energia renovável. As cascas de café são mais adequadas para a produção de gás com rendimento de 40,4%, enquanto que as cascas de sementes de cacau para biocombustível, com rendimento de 37,4%. Até 2021, o potencial energético teórico calculado é de 8291 MWh equivalente a 716000 kg para cascas de café e 1384 MWh equivalente a 121000 kg para cascas de sementes de cacau. A caracterização de bio-carvão indicou altos teores de carbono, cálcio, potássio, nitrogénio e compostos oxigenados, o que os torna candidatos ideais para uso como fertilizantes do solo. A conversão da casca do café e a casca das sementes de cacau em produtos de maior valor agregado, através do processo de pirólise lenta, ajudará a manter o um ambiente limpo, reduzirá o efeito estufa e contribuirá com renda adicional para os agricultores das populações rurais.

Monomorphic pathogens: The case of Candidatus Xenohaliotis californiensis from abalone in California, USA and Baja California, Mexico.

Withering syndrome (WS) is a chronic wasting disease affecting abalone species attributed to the pathogen Candidatus Xenohaliotis californiensis (CXc). Wild populations of blue (Haliotis fulgens) and yellow (H. corrugata) abalone have experienced unusual mortality rates since 2009 off the peninsula of Baja California and WS has been hypothesized as a possible cause. Currently, little information is available about the genetic diversity of CXc and particularly the possible existence of strains differing in pathogenicity. In a recent phylogenetic analysis, we characterized five coding genes from this rickettsial pathogen. Here, we analyze those genes and two additional intergenic non-coding regions following multi-locus sequence typing (MLST) and multi-spacer typing (MST) approaches to assess the genetic variability of CXc and its relationship with blue, yellow and red (H. rufescens) abalone. Moreover, we used 16S rRNA pyrosequencing reads from gut microbiomes of blue and yellow abalone to complete the genetic characterization of this prokaryote. The presence of CXc was investigated in more than 150 abalone of the three species; furthermore, a total of 385 DNA sequences and 7117 16S rRNA reads from Candidatus Xenohaliotis californiensis were used to evaluate its population genetic structure. Our findings suggest the absence of polymorphism in the DNA sequences of analyzed loci and the presence of a single lineage of CXc infecting abalone from California (USA) and Baja California (Mexico). We posit that the absence of genetic variably in this marine rickettsia may be the result of evolutionary and ecological processes.

Thermal effect of acute and chronic stress on hepatic and renal tissue of the pacific sardine, Sardinops sagax caeruleus (Jenyns, 1842) / Efecto del estrés térmico agudo y crónico sobre el tejido hepático y renal de la sardina del pacífico, Sardinops sagax caeruleus (Jenyns, 1842)

SUMMARY: In the thermal range of the Pacific waters, known for the geographical distribution of Sardinops sagax caeruleus(Jenyns, 1842) (Pacific sardine), could be exposed to a stressing temperature environment with dilated effect. This work examines the liver and kidney of sardines acclimated at different temperatures and exposed to the lethal temperature and critical thermal maximum trials. The liver and kidney tissues of Sardinops sagax caeruleus acclimated for 25 days at 19, 21, 23 and 25 °C were anatomically examined after exposure to acute heat stress (AHS) caused by increasing the water temperature at a rate of 1 °C per min, and the chronic heat stress (CHS) effect by abruptly exposure to constant water temperature different from that of acclimation (AT). We observed in fish exposed to AHS that the liver tissue had vacuolated or necrotic hepatocytes and infiltration of inflammatory blood cells (25 °C) and the kidney tissue showed degenerative changes in the glomeruli and renal tubules and increased melanomacrophage centers. The CHS effect in liver and renal tissues produced damage signs of pyknosis, apoptosis, necrotic areas, and an increase in melanomacrophage centers as well as outbreaks of bacterial infection. The results demonstrate that S. sagax caeruleus did not tolerate an abrupt thermal change of more than 4 °C, independently of the ATs, over 50 % died. The consequences of the experimental acute and chronic thermal stress were histopathological alterations of liver and kidney. It was expected that the chronic stress temperature could produce in fish conspicuous histological changes, and indeed it was the most deleterious.

RESUMEN: En el rango térmico de las aguas del Pacífico, conocido por la distribución geográfica de Sardinops sagax caeruleus (Jenyns, 1842) (sardina del Pacífico), esta especie podría estar expuesta a un ambiente de temperatura estresante con efecto dilatado. Este trabajo examina el hígado y el riñón de las sardinas aclimatadas a diferentes temperaturas y expuestas a la temperaturas letales y crítica máxima. Los tejidos hepático y renal de Sardinops sagax caeruleus aclimatadas durante 25 días a 19, 21, 23 y 25 °C se examinaron anatómicamente después de la exposición al estrés de calor agudo (AHS) causado por el aumento de la temperatura del agua a una velocidad de 1 °C por minuto, y el efecto de estrés de calor crónico (CHS) por la exposición brusca a una temperatura constante del agua diferente a la de la aclimatación (AT). Observamos en peces expuestos a AHS que el tejido hepático tenía hepatocitos vacuolados o necróticos y la infiltración de células sanguíneas (25 °C) y el tejido renal presentaba cambios degenerativos en los glomérulos y túbulos renales y aumento de centros de melanomacrófagos. El efecto CHS en los tejidos hepático y renal produjo signos de daño de picnosis, apoptosis, áreas necróticas y un aumento en los centros de melanomacrófagos, así como brotes de infección bacteriana. Los resultados demuestran que S. sagax caeruleus no toleró un cambio térmico abrupto de más de 4 °C, independientemente de las AT, más del 50 % murió. Las consecuencias del estrés térmico agudo y crónico experimental fueron las alteraciones histopatológicas del hígado y el riñón. Se esperaba que la temperatura de estrés crónica pudiera producir cambios histológicos conspicuos en los peces, y de hecho fue la más perjudicial.

Multigenetic characterization of 'Candidatus Xenohaliotis californiensis'.

'Candidatus Xenohaliotis californiensis' (or Ca.Xc) is the aetiological agent of withering syndrome, a chronic wasting disease affecting most if not all North American species of abalone, and has been described as a Rickettsiales-like prokaryote. Genetic data regarding this species are limited to the 16S rRNA gene. The inability to grow it axenically has hindered its genetic and genomic characterization and, in consequence, a thorough analysis of its systematics. Here, we amplified and sequenced five genes (16S rRNA, 23S rRNA, ftsZ, virD4 and virB11) of Ca.Xc from infected abalone to analyse its phylogenetic position. Phylogenies from concatenated DNA and amino acid sequences with representative genera of most Rickettsiales unequivocally place Ca.Xc in the family Anaplasmataceae. Furthermore, the family has two reciprocally monophyletic lineages: one leading to (Neorickettsia, Ca.Xc) and the other to ((Ehrlichia, Anaplasma), Wolbachia)). A molecular-clock Bayesian reconstruction places Ca.Xc as the most basal lineage in Anaplasmataceae. These phylogenetic hypotheses shed light on patterns of host evolution and of ecological transitions. Specifically, Neorickettsia and Ca.Xc inhabit aquatic hosts whereas the remaining Anaplasmataceae are found in terrestrial hosts. Additionally, our evolutionary timeline places the directly transmitted marine Ca.Xc as the basal Anaplasmataceae, ancestral to both freshwater and terrestrial species with adaptations leading to more complex life cycles involving intermediate vectors or reservoir species; this supports the hypothesis of a marine origin for this bacterial family.

Hyperparasitism by the bacteriophage (Caudovirales) infecting Candidatus Xenohaliotis californiensis (Rickettsiales-like prokaryote) parasite of wild abalone Haliotis fulgens and Haliotis corrugata from the Peninsula of Baja California, Mexico.

Candidatus Xenohaliotis californiensis (CXc) is a Rickettsiales-like prokaryote that is considered the causal agent of Withering Syndrome (WS), a chronic disease of abalone, from the west coast of North America and it is listed by the International Organization for Animal Health (OIE) as a reportable agent due to its pathogenicity. This bacterium in red abalone Haliotis rufescens, black abalone Haliotis cracherodii, and yellow abalone Haliotis corrugata from California, US and Baja California, Mexico has been found to be infected by a bacteriophage. To date, there is no information on the epizootiology of CXc and its bacteriophage in natural populations of abalone; furthermore, it is unknown if the bacteriophage was also present in CXc infecting blue abalone Haliotis fulgens. The objective of this study was to determine the distribution, prevalence and intensity of CXc, as well as to determine the distribution and prevalence of the bacteriophage and to study interactions between host sex and hyperparasitism in blue abalone and yellow abalone. Tissue samples were obtained from seven localities where the commercial capture of wild abalone is carried out. Samplings were conducted throughout the 2012-2013 capture seasons and a total of 182 blue abalone and 170 yellow abalone were obtained. The prevalence and intensity of CXc and the prevalence of the bacteriophage were determined by histology. The identity of CXc was confirmed by PCR, product sequence analysis and in situ hybridization while the identity of the bacteriophage was corroborated by TEM. The prevalence of CXc infected and uninfected by the bacteriophage was 80% in blue abalone and 62% in yellow abalone. Low infection intensities were found in 86% of blue abalone and 82% of yellow abalone. Infection intensity was significantly higher in undifferentiated yellow abalone. The bacteriophage in CXc showed a prevalence of 22% and 31% in blue abalone and yellow abalone respectively. These results show that CXc and its bacteriophage are widely distributed in the peninsula of Baja California and that they are well established in natural populations of blue abalone and yellow abalone. Additionally, this data constitutes the first record of a bacteriophage in blue abalone.

Ontogeny of the digestive system of the Octopus bimaculatus paralarvae (Verril, 1883).

The high mortalities registered in the larval stage during octopus culturing are mainly due to nutritional deficiencies of the food provided. To understand the cause of this problem, we studied the ontogenetic development of the digestive system of Octopus bimaculatus paralarvae. An egg batch was obtained from a gravid female collected in the Bay of Los Angeles, Baja California, Mexico, and it was incubated in the laboratory during the summer of 2011. We observed that the formation of the digestive system began at 33 days post-laying (DPL). The newly hatched paralarvae had already formed the organs involved in food ingestion and digestion, although it was not possible to know accurately their degree of maturity. The present research constitutes the first description at the histological level of the ontogenic development of the digestive system of the O. bimaculatus paralarvae. This serves as a basis for future studies of the digestive physiology of this species.