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[Display omitted] • The Rickettsiales-like organism from Crassostrea corteziensis is a commensal endosymbiont. • The prevalence and intensity of the Rickettsiales-like organism is low. • 16S rRNA amplicon sequencing shows the presence of intracellular parasite OTUs. Fastidious endosymbiotic Rickettsiales-like organisms (RLOs) have been observed in the digestive diverticula of the cultured pleasure oyster (Crassostrea corteziensis) from Nayarit, Mexico since 2007. In a few mollusk species, these bacteria have been associated with mortality events and production losses. The type of relationship between the RLOs and the pleasure oyster is largely unknown and further investigations are needed to determine if these bacteria warrant management concern in C. corteziensis. In this study, the morphological characteristics of the RLOs were studied by histology and SEM, and the taxonomic affiliations of the bacteria were evaluated by 16S rRNA amplicon sequencing. In addition, the prevalence and intensity of the RLOs was recorded from 2007 to 2017 by histology. The RLOs were observed inside circular basophilic cytoplasmic membrane bound vacuoles (MBVs) that had an average length and width of 15.70 ± 15.24 µm and 15.42 ± 14.95 µm respectively. Apart from cellular hypertrophy, no tissue alterations were observed in the areas adjacent to the RLOs. Individual bacteria within the MBVs were coccoid in shape with an average length of 0.65 ± 0.12 µm and an average width of 0.38 ± 0.09 µm. The bacterial microbiota of a selected number of samples (one sample without RLOs and two samples with RLOs) showed the presence of intracellular parasite OTUs corresponding to the families Rickettsiaceae and Anaplasmataceae, suggesting that the RLOs from the pleasure oyster is associated with the order Rickettsiales. A mean prevalence of 5 % was observed throughout the study period and the majority of the organisms (89 %) presented low intensity of Grade 1 (30–61 RLOs) of the MBVs. A higher prevalence of the RLOs was observed during warmer months. The lack of tissue alterations, the low prevalence and the low intensity of the MBVs suggest that the RLOs from C. corteziensis is a commensal endosymbiont that presents little risk for oyster production in Nayarit, México. However, regular monitoring is needed to detect if any variation in this relationship occurs, mainly in a scenario where extreme environmental fluctuations may occur. [ABSTRACT FROM AUTHOR]

Bacteriophages are recognized as major mortality agents of microbes, among them intracellular marine rickettsiales-like bacteria. Recently, a phage hyperparasite of Candidatus Xenohaliotis californiensis (CXc) has been described. This bacterium is considered the causal agent of Withering Syndrome (WS) which is a chronic and potentially lethal disease of abalone species from California, USA and the peninsula of Baja California, Mexico. This hyperparasite which infects CXc could be used as a biocontrol agent for WS. Therefore, it is necessary to obtain genomic information to characterize this phage. In this study, the first complete genome sequence of a novel phage, Xenohaliotis phage (pCXc) was determined. The complete genome of pCXc from red abalone (Haliotis rufescens) is 35,728 bp, while the complete genome of pCXc from yellow abalone (Haliotis corrugata) is 35,736 bp. Both phage genomes consist of double-stranded DNA with a G + C content of 38.9%. In both genomes 33 open reading frames (ORFs) were predicted. Only 10 ORFs encode proteins that have identifiable functional homologues. These 10 ORFs were classified by function, including structural, DNA replication, DNA packaging, nucleotide transport and metabolism, life cycle regulation, recombination and repair, and additional functions. A PCR method for the specific detection of pCXc was developed. This information will help to understand a new group of phages that infect intracellular marine rickettsiales-like bacteria in mollusks. [ABSTRACT FROM AUTHOR]

Candidatus Xenohaliotis californiensis ( C Xc) 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 C Xc and its bacteriophage in natural populations of abalone; furthermore, it is unknown if the bacteriophage was also present in C Xc infecting blue abalone Haliotis fulgens . The objective of this study was to determine the distribution, prevalence and intensity of C Xc, 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 C Xc and the prevalence of the bacteriophage were determined by histology. The identity of C Xc was confirmed by PCR, product sequence analysis and in situ hybridization while the identity of the bacteriophage was corroborated by TEM. The prevalence of C Xc 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 C Xc showed a prevalence of 22% and 31% in blue abalone and yellow abalone respectively. These results show that C Xc 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. [ABSTRACT FROM AUTHOR]

The protozoan parasite Perkinsus marinus is the etiological agent of “dermo disease”. This pathogen is considered by the World Organization for Animal Health (OIE) as reportable due to the high mortalities that it produces in the eastern oyster Crassostrea virginica in the US. In 2006, this parasite was detected in the pleasure oyster Crassostrea corteziensis in Nayarit on the Pacific coast of Mexico, indicating a new host and an extension of its known distribution. Epizootiological data of P. marinus in the pleasure oyster are unknown. With the objective of determining the prevalence and intensity in relation with temperature and salinity throughout time, as well as for studying interactions of host size and sex with the parasite, a monthly sampling was carried out in two aquaculture sites of Nayarit from 2007 to 2014. A total of 7700 oysters were analyzed. In both localities, prevalence was low in winter (<6%) when temperature and salinity fluctuated around 24 °C and 33, respectively; and the highest prevalence values occurred during summer (37%) when temperature and salinity were around 30 °C and 20, respectively. Infection intensity increased in summer, but severe cases remained on average <10%. Larger oysters showed the highest prevalence and intensity, and higher prevalence were generally observed in females. No unusual mortalities directly related with P. marinus were observed. [ABSTRACT FROM AUTHOR]

Transmission electron microscopy analysis (TEM) of the rickettsiales-like prokaryote, Candidatus Xenohaliotis californiensis ( C Xc), pathogen of Haliotis spp. from the West Coast of North America, were found to be infected by a bacteriophage hyperparasite previously described in red abalone from California. The hyperparasite has an icosahedrical-like capsid with a narrow long flexible tail, this morphological characteristic tentatively place this virus in the Family Siphoviridae from the order Caudovirales . TEM images also showed the bacteriophage in different stages of assembly in the cytoplasm of C Xc, demonstrating its lytic cycle. [ABSTRACT FROM AUTHOR]

Filtrations were applied to separate vacuoles of Rickettsiales-like organism Candidatus Xenohaliotis californiensis. Vacuoles were visualized by staining with nucleic acid fluorochrome and their identity was confirmed by Laser Capture Microdissection and PCR. This methodology separates vacuoles and allows studies without the need of isolation in synthetic media or cell lines. [ABSTRACT FROM AUTHOR]

Abstract: The mangrove oyster Saccostrea palmula coexists with the pleasure oyster Crassostrea corteziensis in coastal lagoons of northwest Mexico. Recent discovery of Perkinsus marinus infecting the pleasure oyster in the region prompted evaluation of S. palmula as an alternative P. marinus host. An analysis to determine the possible presence of P. marinus in natural and cultured populations of S. palmula at four coastal lagoons in Sinaloa, Mexico was carried out during October–November 2010. Tissues from apparently healthy S. palmula were evaluated using Ray’s fluid thioglycollate method (RFTM), which revealed a Perkinsus sp. to be present in all four locations at 6.7–20.0% prevalence. Histopathological analysis of these specimens showed tissue alterations and parasite forms consistent with moderate P. marinus infection, which was confirmed by ribosomal non-transcribed spacer (NTS)-based PCR assays on DNA samples from oysters positive by RFTM and histology. DNA sequencing of amplified NTS fragments (307bp) produced a sequence 98–100% similar to GenBank-deposited sequences of the NTS from P. marinus. Fluorescent in situ hybridization for Perkinsus spp. and P. marinus corroborated the PCR results, showing clear hybridization of P. marinus in host tissues. This is the first record of P. marinus infecting a species from genus Saccostrea and the first record of the parasite from coastal lagoons in Sinaloa, Mexico. [Copyright &y& Elsevier]

Abstract: The blue mussel Mytilus galloprovincialis is cultured in Bahía de Todos Santos where it sustains a growing economic activity. This culture depends on seed collection from artificial collectors in the environment. It is known that the settlement season takes place from late autumn to early winter. However, there is no information on the settlement in different points of the culture area or at different depths. Neither is there knowledge on size composition of the settling population under these conditions. This information is useful for improving seed collection practices and for improving the understanding of the substrate colonization. For obtaining this information, pieces of about 105cm2 of synthetic fibrous material were used as collectors and were placed at 2, 5, 7, 9, 10, 11, 13, 15 and 18m depth in three points of the culture area during the settlement season. Additionally, an analysis of size composition of the settling population was carried out. Major settlement occurred in January in the three studied points. Settlement decreased according to the orientation of the sampling point with respect to the current from West to East, and according to depth. Higher values were found at 2m depth and lower values were found at 18m depth in all the three points. Settlement in February dropped nearly 43% with respect to January and it was relatively uniform in the 3 studied points. Size composition of the settling population shows that the greatest percentage (98%) settled directly as competent larvae and only 2% corresponded to drifting post-larvae. Around 40% of the seed was collected below 7m depth; thus, mussel growers could place their collectors between all the studied depths for catching more seed. [Copyright &y& Elsevier]

Burrowing worms that belong to Polydora spp. infest marine mollusks cultured worldwide, causing problems for production and marketing. The clam Chione fluctifraga is semi-cultured in Bahı´a Falsa, Baja California, NW Mexico, and some clams harbor burrowing worms. The present study was carried out to determine the identity of the worm species infesting the clam, the infesting process by cohabitation of infested and non-infested clams in aquaria with a variety of substrates (fine sand, gross sand, plastic bag used for clam culture, and aquarium without substrate) and turbulence conditions, and the occurrence of architomy phenomena in connection with infestation of the clam. The burrowing worm was considered as a nova species due to its singular limbate neurosetae and notosetae in the setiger 5, hooks in the setiger 6, eyes not present, and general pigmentation, among other characteristics. Infestation was similar in all substrates and turbulence conditions, but it was more abundant on clams previously infested than on those free of worms, showing a preferential settlement of worm infesting stages on pre-infested clams. Regeneration was observed in all segments of the worm: anterior (metastomium), medium, and posterior (prostomium); the complete regeneration time occurred in 40 days. This is the first record of architomy in a species of Polydora and this phenomenon could account for the increase of infestation intensity in pre-infested clams at the end of the study period. Infestation of clams by settling polichaete in the conditions studied, and the architomy process in this worm species, shows its great infesting capacity. [Copyright &y& Elsevier]

[Display omitted] • Dark Leathery Surface of Geoduck Clams (LSGC) was diagnosed in Panopea globosa. • A histophagous protozoan , Uronema marinum, was associated with LSGC in Panopea spp. • Our data constitute an extension in the distribution range of LSGC in North America. Dark Leathery Surface of Geoduck Clams (LSGC) is an alteration that affects the periostracum of the mantle and siphon of Panopea generosa from the northwest coast of Canada and Mexico. This alteration affects commercialization and possibly the survival of the clams. The cause of LSGC is unknown but has been correlated with presence of fungi and protozoans. We detected a similar alteration in Panopea globosa from Baja California, Mexico and the histophagous ciliate Uronema marinum was isolated from affected siphon tissue. U. marinum was identified by its morphology and by genetic analysis of the gene 18S rRNA. This is the first record of LSGC in P. globosa and the first identification of a histophagous protozoan associated with it. [ABSTRACT FROM AUTHOR]

Candidatus Xenohaliotis californiensis (C Xc) is a Rickettsiales-like organism (RLO) that is considered the causal agent of withering syndrome (WS), a chronic and potentially fatal disease for several abalone species. C Xc can be infected by a bacteriophage (p C Xc), and both phage-infected and non-phage-infected C Xc can coexist with a presumable different species of RLO tentatively named stippled Rickettsiales-like organism (ST-RLO). Currently, the taxonomic affiliation of the ST-RLO is unknown, and no studies have been conducted to determine its presence in infected abalones in the Peninsula of Baja California, Mexico. To determine whether the ST-RLO is a histological variant of C Xc or a new species of RLO, we performed histology and species-specific in situ hybridization (ISH) for C Xc and p C Xc in red abalone, Haliotis rufescens , infected with C Xc and hyperparasitized by p C Xc, which also showed the presence of the ST-RLO. Additionally, a histological analysis of the presence of all of these RLOs from 2008 to 2019 from an aquaculture facility was performed. The results showed that all ST-RLOs were negative by ISH for C Xc and p C Xc, suggesting that the ST-RLO is a different species of bacterium and that it is not susceptible to infection by p C Xc. Histologically similar alterations to those produced by C Xc in the post esophagus were observed. The three RLOs regularly coexist in all the infected abalones, and there is a relatively constant proportion of each kind of RLO over time, suggesting that equilibrium is reached during coexistence; the largest percentage corresponded to C Xc non phage infected (81%), followed by C Xc infected by p C Xc (16%) and the ST-RLO (3%). In all cases, the ST-RLO infection remained at a subclinical level making it difficult to determine if it acts as pathogen or as a commensal. Therefore, the genetic characterization of the ST-RLO and pathogenicity studies are needed to further elucidate the role of this new RLO in the development of of WS in abalone species of North America. • Confirmation of the presence of an unidentified rickettsia in abalone. • This unidentified rickettsia produces similar damages to those attributed to Candidatus Xenohaliotis californiensis. • This rickettsia coexists with Candidatus Xenohaliotis californiensis non infected and infected by the bacteriophage pCXc. • The role of this rickettsia in the development of abalone Withering Syndrome is unknown. • This is the first study to perform ISH for pCXc and to confirm that the ST-RLO is not susceptible to infection by pCXc. [ABSTRACT FROM AUTHOR]

• Argovit™ silver nanoparticles show dose-dependent behavior after 24 h of exposure. • The nanoparticles cause severe damage to parasite morphology. • Nanoparticles penetrated the parasite cell wall and degraded lipid droplets. • Nanoparticles maintain their lethal effect for up to 24 h in a saline medium. Perkinsus marinus , a World Organisation for Animal Health (OIE) notifiable parasite, infects several species of oyster, including Crassostrea virginica and Crassostrea corteziensis. There is little information on possible treatments for this parasite, but the biocidal properties of silver nanoparticles (AgNP) suggest their potential use. The lethal effects of the Argovit™ formulation of AgNP was evaluated for the first time against hypnospores of P. marinus , a particularly resistant stage of the parasite that persists in the environment until favorable conditions occur for zoosporulation to be induced. Hypnospores were exposed to 1, 10 and 100 µg/mL of silver compounded in Argovit™ (corresponding to 0.009, 0.093 and 0.927 mM of Ag), to 157.47 µg/mL (0.927 mM) of silver nitrate (AgNO 3) used as a positive control, and to polyvinylpyrrolidone (PVP, 1570 µg/mL) used as a vehicle control. Hypnospores in culture medium without treatment served as a negative control. Dose-dependence after 24 h of exposure to AgNP was observed. A concentration of 0.093 mM AgNP resulted in 50% mortality of P. marinus. Treatment with 0.927 mM of silver, as AgNP or AgNO3, was highly lethal, with greater than 90% mortality. Silver nanoparticles were implicated in the deformation of hypnospores. Transmission electron microscopy (TEM) revealed AgNP within the hypnospore wall and involved in the degradation of lipid droplets in the cytoplasm. AgNP were effective in a saline medium, suggesting the utility of detailed studies of the physicochemical interactions of AgNP under these conditions. These results suggest investigations of possible effect of Argovit™ formulation of AgNP against stages of the parasite like trophozoites and tomonts that develop in tissues or hemolymph of infected oysters as well as studies on its effects in the host and environment. [ABSTRACT FROM AUTHOR]

Unfortunately, the family name of the co-author Roberto Cruz-Flores was incorrectly published in the original publication and corrected here by this correction. The original article has been corrected. [ABSTRACT FROM AUTHOR]

Withering syndrome (WS) is a chronic wasting disease affecting abalone species attributed to the pathogen Candidatus Xenohaliotis californiensis ( C Xc). 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 C Xc 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 C Xc 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 C Xc 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 C Xc 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. [ABSTRACT FROM AUTHOR]

Formalin-fixed paraffin-embedded (FFPE) tissues stored in thousands of human and animal pathology laboratories around the globe represent mines of stored genetic information. In recent years, the use of FFPE tissues as a viable source of DNA for diverse genetic studies has attracted attention for interrogating microbiomes from this sample type. These studies have proven that 16S rRNA amplicon sequencing-based microbiome studies are possible from FFPE samples but present some particular challenges. In this review, we summarize all aspects of microbiome studies from FFPE tissues including the challenges associated with working highly degraded DNA, best practices for reducing environmental contamination, and we propose solutions to address these issues. Finally, we discuss how the combination of FFPE microbiome studies and Laser Capture Microdissection and/or Laser Microdissection could enable to determine the spatial heterogeneity underlying complex bacterial communities. [Display omitted] [ABSTRACT FROM AUTHOR]

Coral reefs are threatened by a variety of factors including diseases that have caused significant damage in some regions such as in the Caribbean. At present, no data are available on coral diseases in the Mexican Pacific where Pocillopora spp. is a dominant component of coral communities. Here, we describe gross and microscopic morphology of lesions found in pocilloporids at four sites in the Mexican Pacific. Corals were identified and their lesions photographed and quantified in the field. Tissue samples were collected from healthy and affected colonies for histopathology. We recorded seven species of pocilloporids at the study sites with Isla Isabel being the location with the highest coral diversity (H' = 1.27). Lesions were present in 42% of the colonies and included discoloration (32%), predation-induced tissue loss (30%), unexplained tissue loss (3%) and overgrowth by sponges or algae (35%). The most affected species, P. damicornis (50%), was also one of the most common in the region. No species was more prone to a particular lesion, but there was a significant association between location and the presence of lesions. Northern Islas Marietas (61%) and Isla Isabel (41%) had the highest prevalence of lesions, followed by Manzanillo (37%) and Bahías de Huatulco (23%). Histological changes included atrophy of the surface body wall with depletion of zooxanthellae (91%) in corals with discoloration (bleaching). Ablation of tissue from mesoglea (18%) was also observed. Colonies with unexplained tissue loss showed atrophy and thinning of the epidermis (89%), characterized by cuboidal instead of pseudocolumnar cells normally found in healthy pseudocolumnar ciliated epithelium. Bacterial aggregates between the mesoglea and gastrodermis (11%) were very conspicuous in healthy and diseased corals. Lesions produced by fish bites and gastropods were associated with tissue atrophy (40%) and, in some cases, algal overgrowth near the lesion (20%). No infectious agents associated with cell pathology were detected microscopically. Bleaching and overgrowth by algae and sponges, as well as unexplained tissue loss, are common in Pocillopora. These lesions and anatomical changes warrant further study since their incidence is potentially indicative of reef degradation. [ABSTRACT FROM AUTHOR]