Phlebotomine (Diptera: Psychodidae) species and their blood meal sources in a new leishmaniasis focus in Los Montes de María, Bolívar, in northern Colombia / Especies de flebotomíneos y sus fuentes de ingesta sanguínea en un nuevo foco de leishmaniasis en Los Montes de María (Bolívar) al norte de Colombia.

Introduction. El Alférez, a village in Los Montes de María (Bolívar, Colombia) and a macro-focus of leishmaniasis, recorded its first case in 2018, evidencing changes in the distribution and eco-epidemiology of the disease, although interactions between vectors and local fauna remain unknown. Objective. To evaluate the diversity of sandflies and their blood meal sources in the community of El Alférez in the municipality of El Carmen de Bolívar (Bolívar, Colombia). Materials and methods. In 2018, sandflies were collected using LED-based light traps in domestic, peridomestic, and sylvatic ecotopes and identified at the species level. Multiplex polymerase chain reaction targeting the mitochondrial cytochrome B gene was used to analyze blood from the digestive tract. Results. Lutzomyia evansi was the most abundant species (71.85%; n = 485/675), followed by Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci, and Lu.aclydifera. Twenty-five percent of the species had blood meals from Canis familiaris (36.00%; n = 9/25), Ovis aries (36.00%; n=9:/25), Bos taurus (24.00%; n = 6/25), Sus scrofa (20.00%; n = 5/25), and Homo sapiens (8.00%; n = 2/25). Lutzomyia evansi registered the highest feeding frequency (68.00%; n = 17/25), predominantly on a single (44.00%; n = 11/25) or multiple species (24.00%; n = 6/25). Conclusion. Results indicate a eclectic feeding behavior in Lu. evansi, implying potential reservoir hosts for Leishmania spp. and increasing transmission risk. This study is a first step towards understanding the diversity of mammalian blood sources used by sandflies, that may be crucial for vector identification and formulation of effective control measures.

Introducción. En 2018, en la vereda El Alférez de Los Montes de María (Bolívar, Colombia), un macrofoco de leishmaniasis, se reportó el primer caso y se evidenciaron cambios en la distribución y ecoepidemiología de la enfermedad. No obstante, las interacciones entre vectores y fauna local aún son desconocidas. Objetivo. Evaluar la diversidad de flebotomíneos y sus fuentes de alimentación sanguínea en la comunidad de El Alférez del municipio de El Carmen de Bolívar (Bolívar, Colombia). Materiales y métodos. En el 2018, se recolectaron flebotomíneos mediante trampas de luz led ubicadas en el domicilio, el peridomicilio y en el área silvestre, y se identificaron a nivel de especie. Se utilizó la reacción en cadena de la polimerasa múltiple dirigida al gen mitocondrial citocromo B para analizar la sangre del aparato digestivo. Resultados. Lutzomyia evansi fue la especie más abundante (71,85 %; n = 485/675), seguida por Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci y Lu. aclydifera. El 25 % (n = 25/100) de las especies analizadas tuvieron como fuentes de ingesta sanguínea a Canis familiaris (36 %; n = 9/25), Ovis aries (36 %; n = 9/25), Bos taurus (24 %; n = 6/25), Sus scrofa (20 %; n = 5/25) y Homo sapiens (8 %; n = 2/25). Lutzomyia evansi fue la especie con la mayor frecuencia de alimentación (68 %; n = 17/25), predominantemente de una sola especie (44 %; n = 11/25) o de varias (24 %; n = 6/25).

Integrative Taxonomy Suggests Resurrection of Species of the Astyanax bimaculatus Group (Characiformes, Characidae).

Using integrative tools can be effective for species identification, especially in complex groups like Astyanax. Astyanax bimaculatus group is composed of six valid species, including A. lacustris. "A. altiparanae", "A. asuncionensis", and "A. jacuhiensis" are considered as junior synonyms of A. lacustris. Seeking to test the operational taxonomic unit (OTU) status of the junior synonyms of A. lacustris ("A. altiparanae", "A. asuncionensis", and "A. jacuhiensis"), we used analyses through mitochondrial DNA (COI and Cytb), cytogenetic markers (classical and molecular), and morphometry ("truss network"). Analysis of mitochondrial DNA sequences separated A. lacustris from the other synonymized species. The cytogenetic and morphometric analyses did not corroborate the synonymization and suggest that besides A. lacustris, the OTUs A. altiparanae, A. asuncionensis, and A. jacuhiensis are valid species. The analysis of different characters proposed by the integrative taxonomy used on the same individuals could provide greater reliability and minimize the underestimation of biodiversity.

Molecular based identification and phylogenetic relationship by using cytochrome b gene of Pangasius pangasius / Identificação de base molecular e relação filogenética usando o gene citocromo b de Pangasius pangasius

Molecular appraoch for identification of unknown species by using Cytochrome b gene is an effective and reliable as compared with morphological based identification. For DNA barcoding universal molecular genes were used to identify the species. Cytochrome b is a specific gene used for identification purpose. DNA barcoding is a reliable and effective method compared to the different traditional morphological methods of specie identification. So,in the present study which was conducted to identify the species, a total of 50 fish samples were collected from five different sites. DNA was extracted by using the Phenol Chloroform method from muscle tissue. Five sequences were sequenced (one from each site), analyzed, and identified specific species as Pangasius pangasius. Identified sequences were variable in length from 369 bp (Site 1), 364 bp (Site 2), 364 bp (Site 3), 352 bp (Site 4), and 334 bp (Site 5). Identity matches on the NCBI database confirmed the specific specie as P. pangasius. A distancing tree was drawn to show maximum likelihood among the same and different species. Yet, in many cases fishes on diverse development stages are difficult to identify by morphological characters. DNA-based identification methods offer an analytically powerful addition or even an alternative tool for species identification and phylogenetic study. This work intends to provide an updated and extensive overview on the DNA based methods for fish species identification by using Cytochrome b gene as targeted markers for identification purpose.

A abordagem molecular para identificação de espécies desconhecidas usando o gene citocromo b é eficaz e confiável em comparação com a identificação baseada na morfologia. Códigos de barras de DNA de genes moleculares universais foram usados ​​para identificar as espécies. O citocromo b é um gene específico usado para fins de identificação. O código de barras de DNA é um método confiável e eficaz em comparação com os diferentes métodos morfológicos tradicionais de identificação de espécies. Assim, no presente estudo, que foi realizado para identificar as espécies, um total de 50 amostras de peixes foram coletadas em cinco locais diferentes. O DNA foi extraído usando o método Fenol Clorofórmio do tecido muscular. Cinco sequências foram sequenciadas (uma de cada local), analisadas e identificadas espécies específicas, como Pangasius pangasius. As sequências identificadas tinham comprimento variável de 369 bp (Local 1), 364 bp (Local 2), 369 bp (Local 1), 364 bp (Local 3), 352 bp (Local 4) e 334 bp (Local 5). As correspondências de identidade no banco de dados do NCBI confirmaram a espécie específica como P. pangasius. Uma árvore de distanciamento foi desenhada para mostrar a máxima probabilidade entre elas e diferentes espécies. No entanto, em muitos casos, peixes em diversos estágios de desenvolvimento são difíceis de identificar por caracteres morfológicos. Os métodos de identificação baseados em DNA oferecem uma adição analiticamente poderosa ou mesmo uma ferramenta alternativa para identificação de espécies e estudo filogenético. Este trabalho pretende fornecer uma visão geral atualizada e abrangente sobre os métodos baseados em DNA para identificação de espécies de peixes usando o gene citocromo b como marcadores direcionados para fins de identificação.

Hexa-Histidine, a Peptide with Versatile Applications in the Study of Amyloid-ß(1-42) Molecular Mechanisms of Action.

Amyloid ß (Aß) oligomers are the most neurotoxic forms of Aß, and Aß(1-42) is the prevalent Aß peptide found in the amyloid plaques of Alzheimer's disease patients. Aß(25-35) is the shortest peptide that retains the toxicity of Aß(1-42). Aß oligomers bind to calmodulin (CaM) and calbindin-D28k with dissociation constants in the nanomolar Aß(1-42) concentration range. Aß and histidine-rich proteins have a high affinity for transition metal ions Cu2+, Fe3+ and Zn2+. In this work, we show that the fluorescence of Aß(1-42) HiLyteTM-Fluor555 can be used to monitor hexa-histidine peptide (His6) interaction with Aß(1-42). The formation of His6/Aß(1-42) complexes is also supported by docking results yielded by the MDockPeP Server. Also, we found that micromolar concentrations of His6 block the increase in the fluorescence of Aß(1-42) HiLyteTM-Fluor555 produced by its interaction with the proteins CaM and calbindin-D28k. In addition, we found that the His6-tag provides a high-affinity site for the binding of Aß(1-42) and Aß(25-35) peptides to the human recombinant cytochrome b5 reductase, and sensitizes this enzyme to inhibition by these peptides. In conclusion, our results suggest that a His6-tag could provide a valuable new tool to experimentally direct the action of neurotoxic Aß peptides toward selected cellular targets.

Phylogenetic relationships, distribution, and conservation of Roosmalens' dwarf porcupine, Coendouroosmalenorum Voss & da Silva, 2001 (Rodentia, Erethizontidae).

The New World porcupines of the genus Coendou comprise 16 species of arboreal nocturnal rodents. Some of these species are poorly known and have not been included in phylogenetic analyses. Based on recently collected specimens with associated tissue from the Brazilian Amazonia, we investigate the distribution and phylogenetic relationships of Roosmalens' dwarf porcupine, Coendouroosmalenorum, using an integrative approach using mitochondrial gene sequences and morphological data from new specimens and localities. Our results recovered C.roosmalenorum in the subgenus Caaporamys. However, analyses of our molecular and combined datasets produced different topologies. The new record shows the presence of C.roosmalenorum 480 km to the southeast of the Rio Madeira and 95 km away from Rio Juruena in Mato Grosso state, indicating a wider distribution in southern Amazonia than suspected. All known records of C.roosmalenorum are in the Madeira biogeographical province, to which it might be endemic.

Survey of phlebotomine sand fly fauna in a public Zoo in Brazil: Species diversity, seasonality, and host variety.

Leishmaniasis is a dynamic disease in which transmission conditions change due to environmental and human behavioral factors. Epidemiological analyses have shown modifications in the spread profile and growing urbanization of the disease, justifying the expansion of endemic areas and increasing number of cases in dogs and humans. In the city of Belo Horizonte, located in the southeastern state of Minas Gerais (Brazil), visceral leishmaniasis (VL) is endemic, with a typical urban transmission pattern, but with different regional prevalence. This study was conducted at the Zoo of the Foundation of Municipal Parks and Zoobotany of Belo Horizonte (FPMZB-BH), located in the Pampulha region, which is among the areas most severely affected by VL. This study aimed to determine the taxonomic diversity of native phlebotomine sand flies (Diptera: Psychodidae), identify climatic variables that potentially affect the phenology of these insects, and determine the blood meal sources for female phlebotomine sand flies. To achieve this, 10 mammal enclosures in the zoo were selected using the presence of possible leishmaniasis reservoirs as a selection criterion, and sampled using light traps between August 2019 and August 2021. A total of 6034 phlebotomine sand flies were collected, indicating nine species, with Lutzomyia longipalpis being the very abundant species (65.35% of the total). Of the 108 engorged phlebotomine collected females, seven samples (6.5%) were positive for blood meals from humans, marsupials, canids, and birds. Relative humidity and rainfall increased the phenology of phlebotomine sand flies, with population increases in the hottest and wettest months. The data obtained will provide guidelines for competent health agencies to implement vector control measures to reduce the risk of leishmaniasis transmission in the FPMZB-BH.

The recent introduction of Angiostrongylus cantonensis and its intermediate host Achatina fulica into Guadeloupe detected by phylogenetic analyses.

BACKGROUND: Angiostrongylus cantonensis (rat lungworm) is the main pathogen responsible for eosinophilic meningitis in humans. One of its intermediate snail hosts, Achatina fulica, was already present in many countries around the world before it appeared in the West Indies in the late 1980s. In the French territories in the Caribbean and northern South America, the first cases of human neuroangiostrongyliasis were reported in Martinique, Guadeloupe and French Guiana in 2002, 2013 and 2017, respectively. In order to better characterize angiostrongyliasis in Guadeloupe, particularly its geographical origin and route of introduction, we undertook molecular characterization of adult worms of Angiostrongylus cantonensis and its intermediate host Achatina fulica. METHODS: Genomic DNA of adult Angiostrongylus cantonensis and Achatina fulica was extracted and amplified by polymerase chain reaction (PCR) targeting the mitochondrial genes cytochrome B and C for A. cantonensis and 16S ribosomal RNA for A. fulica. The PCR products were sequenced and studied by phylogenetic analysis. RESULTS: Cytochrome B and cytochrome C molecular markers indicate a monophyletic lineage of A. cantonensis adult worms in Guadeloupe. Two sequences of A. fulica were identified. CONCLUSIONS: These results confirm the recent introduction of both Angiostrongylus cantonensis and Achatina fulica into Guadeloupe. Achatina fulica in Guadeloupe shares a common origin with those in Barbados and New Caledonia, while Angiostrongylus cantonensis in Guadeloupe shares a common origin with those in Brazil, Hawaii and Japan.

Morphological and molecular analyses confirm the presence of the newly described bat species Molossus melini in the Espinal ecoregion from Argentina.

Until now, Molossus melini was known only from its type locality, in the south of Santa Fe province, Argentina. Specimens of this species were collected in 2021 from a roost in a Fraxinus tree of the urban woodland of Paraná city, Entre Ríos province, Argentina. Bat identification was made by comparing external and cranial characters and measurements with those reported in the bibliography and corroborated by a phylogenetic analysis based on the cytochrome b gene. Also, multivariate morphometric analyses showed that cranial measurements, but not external ones, are informative enough to discriminate M. melini from the other Molossus species in Argentina (M. currentium, M. fluminensis, and M. molossus). This new record extends the distribution of M. melini from the south of Santa Fe province by 230 km to the northeast and represents the first record of the species in the Espinal ecoregion. Supplementary information: The online version contains supplementary material available at 10.1007/s13364-023-00679-1.

Genetic characterization of Dibothriocephalus latus and Dibothriocephalus dendriticus (Cestoda: Diphyllobothriidae) from Chile based on haplotype analysis using mitochondrial DNA markers.

Dibothriocephalus latus and Dibothriocephalus dendriticus are found throughout the temperate and sub-arctic zones of the northern hemisphere, but they are also found in the southern core countries of South America, Chile and Argentina. Genetic characteristics of D. latus and D. dendriticus from South America have yet to be fully defined. The present study aimed to understand the genetic characteristics of D. latus and D. dendriticus from Chile by haplotype network analysis of mitochondrial cytochrome c oxidase subunit I gene (cox1) and cytochrome b gene (cob), as well as their origins. Dibothriocephalus latus and D. dendriticus plerocercoid larvae were obtained from feral and/or wild salmonids captured in Lake Llanquihue in Región de Los Lagos, and Lake Panguipulli in Región de Los Ríos, located south of central Chile. Haplotype analysis of D. latus revealed that H1 in cox1 and H2 in cob are the key haplotypes common to D. latus across the world, including Chile, and both genes exhibited limited genetic diversity in D. latus. It was assumed that D. latus was brought into South America by European and Russian immigrants in the 19th century as previously reported. In contrast, both the cox1 and cob of D. dendriticus display considerable genetic diversity, with no common haplotypes between D. dendriticus populations from Chile and the northern hemisphere. More intriguingly, two cob haplotypes (H24, H25) detected in Chilean D. dendriticus were closely linked to haplotypes (H30, H31) detected in North American D. dendriticus, strongly implying that D. dendriticus in Chile was brought by piscivorous migrating birds from North America. It has also been estimated that the D. dendriticus from Chile genetically diverged from the D. dendriticus from the northern hemisphere approximately 1.11 million years ago, long before humans migrated to the southern parts of South America.

Cytotaxonomy and Molecular Analyses of Mycteria americana (Ciconiidae: Ciconiiformes): Insights on Stork Phylogeny.

Although molecular information for the wood stork (Mycteria americana) has been well described, data concerning their karyotypical organization and phylogenetic relationships with other storks are still scarce. Thus, we aimed to analyze the chromosomal organization and diversification of M. americana, and provide evolutionary insights based on phylogenetic data of Ciconiidae. For this, we applied both classical and molecular cytogenetic techniques to define the pattern of distribution of heterochromatic blocks and their chromosomal homology with Gallus gallus (GGA). Maximum likelihood analyses and Bayesian inferences (680 bp COI and 1007 bp Cytb genes) were used to determine their phylogenetic relationship with other storks. The results confirmed 2n = 72, and the heterochromatin distribution pattern was restricted to centromeric regions of the chromosomes. FISH experiments identified fusion and fission events involving chromosomes homologous to GGA macrochromosome pairs, some of which were previously found in other species of Ciconiidae, possibly corresponding to synapomorphies for the group. Phylogenetic analyses resulted in a tree that recovered only Ciconinii as a monophyletic group, while Mycteriini and Leptoptlini tribes were configured as paraphyletic clades. In addition, the association between phylogenetic and cytogenetic data corroborates the hypothesis of a reduction in the diploid number throughout the evolution of Ciconiidae.

Surveillance of avian malaria and related haemoparasites in common terns (Sterna hirundo) on the Atlantic coast of South America.

Haemosporidia (Apicomplexa, Haemosporida) are protozoa that infect vertebrate blood cells and are transmitted by vectors. Among vertebrates, birds possess the greatest diversity of haemosporidia, historically placed in 3 genera: Haemoproteus, Leucocytozoon and Plasmodium, the causative agent of avian malaria. In South America, existing data on haemosporidia are spatially and temporally dispersed, so increased surveillance is needed to improve the determination and diagnosis of these parasites. During the non-breeding season in 2020 and 2021, 60 common terns (Sterna hirundo) were captured and bled as part of ongoing research on the population health of migratory birds on the Argentinian Atlantic coast. Blood samples and blood smears were obtained. Fifty-eight samples were screened for Plasmodium, Haemoproteus and Leucocytozoon, as well as for Babesia parasites by nested polymerase chain reaction and by microscopic examination of smears. Two positive samples for Plasmodium were found. The cytochrome b lineages detected in the present study are found for the first time, and are close to Plasmodium lineages found in other bird orders. The low prevalence (3.6%) of haemoparasites found in this research was similar to those reported for previous studies on seabirds, including Charadriiformes. Our findings provide new information about the distribution and prevalence of haemosporidian parasites from charadriiforms in the southernmost part of South America, which remains understudied.

Taxonomic status of Tamarinus imperator subgrisescens (Lönnberg, 1940) (Cebidae, Callitrichinae)

The emperor tamarin, Tamarinus imperator, is composed of two subspecies, the nominal type, T. i. imperator, distribut-ed between the Acre and Purus Rivers, whose range is limited between the Brazilian state of Acre and Peru are unbounded, and T. i. subgrisescens, occurring in Peru, Bolivia, and Brazil, in the Brazilian states of Acre and Amazonas. Morphologically, both taxa are easily identifiable by the pelage pattern (chromogenetic fields), and even being easily distinguishable, both lineages are con-sidered subspecies according to the criterion based on the Biological Concept of Species from the 1970s, even without presenting some necessary criteria, such as the intergradation zone. Here we analyzed pelage traits, cranial morphometry, Cytochrome-b di-vergence, and distributional pattern data applying the premises of integrative taxonomy to elucidate the taxonomic status of both lineages. We hypothesize that both lineages are considered full species through a series of criteria for species recognition, such as distinguishability, level of phenotypical divergences of several morphological complexes with congruence among them, and some genetic divergence. The hybridization is unknown and the low or the lack of sampling in target areas does not allow us to determine whether a hybridization or even contact zone between the two lineages exists indeed. All character sets analyzed were congruent with each other and reinforced the high level of divergences between the two subspecies including several pelage differences, mor-phometry (descriptive statistics, PCA, and MANOVA), and mitochondrial DNA Cytochrome-b divergence. Most of the distribution in both lineages are allopatric, and the levels of intra-lineage phenotypical variation are much lower than between the lineages.(AU)

Chromosome Evolution of the Liolaemus monticola (Liolaemidae) Complex: Chromosomal and Molecular Aspects.

Chromosomal rearrangements can directly influence population differentiation and speciation. The Liolaemus monticola complex in Chile is a unique model consisting of several chromosome races arranged in a latitudinal sequence of increasing karyotype complexity from south to north. Here, we compared chromosomal and mitochondrial cytochrome b data from 15 localities across the northern geographic distribution of L. monticola. We expanded the distribution of the previously described Multiple Fissions race (re-described as MF2), in the Coastal range between the Aconcagua River and the Petorca River, and described a new Multiple Fissions 1 (MF1) race in the Andean range. Both races present centric fissions in pairs 1 and 2, as well as a pericentric inversion in one fission product of pair 2 that changes the NOR position. Additionally, we detected a new chromosomal race north of the Petorca River, the Northern Modified 2 (NM2) race, which is polymorphic for novel centric fissions in pairs 3 and 4. Our results increase the number of chromosomal races in L. monticola to seven, suggesting a complex evolutionary history of chromosomal rearrangements, population isolation by barriers, and hybridization. These results show the relevant role of chromosome mutations in evolution, especially for highly speciose groups such as Liolaemus lizards.

Importancia clínica del diagnóstico de la metahemoglobinemia / Clinical importance of the diagnosis of methemoglobinemia

Introducción: La metahemoglobina es una forma de hemoglobina en la que el grupo hemo, usualmente en forma ferrosa, es oxidado a forma férrica, lo que afecta el transporte de oxígeno. El incremento por encima de los valores de referencia se denomina metahemoglobinemia. Objetivo: Actualizar conceptos como prevención, manifestaciones clínicas, diagnóstico de laboratorio y tratamiento de elección de esta enfermedad, con la información disponible de la última década. Métodos: Se realizó una revisión de la literatura en inglés y español, a través del sitio web PubMed, el motor de búsqueda Google académico y Scielo, de artículos publicados en los últimos 10 años. Los términos de búsqueda usados incluyeron metahemoglobinemia, déficit de citocromo b5 reductasa, cianosis y cooximetría. Análisis y síntesis de la información: La metahemoglobinemia se puede clasificar en congénita y adquirida, esta última es la más frecuente. Es importante el diagnóstico de esta enfermedad que aunque es un padecimiento poco común, puede cursar con complicaciones graves e incluso la muerte. Puede ser evitable con diagnóstico temprano y tratamiento oportuno para reducir las complicaciones asociadas a este cuadro. Conclusiones: El diagnóstico y el tratamiento, profiláctico y terapéutico de la metahemoglobinemia en su etapa aguda o de mantenimiento, requieren la adecuada actualización del profesional de la salud(AU)

Introduction: Methemoglobin is a form of hemoglobin in which the heme group, usually in the ferrous form, is oxidized to the ferric form, which affects oxygen transport. The increase above the reference values ​​is called methemoglobinemia. Objective: To update concepts such as prevention, clinical manifestations, laboratory diagnosis and treatment of choice for this disease, with the information available from the last decade. Methods: A review of the literature in English and Spanish was carried out, through the PubMed website, the academic Google search engine and Scielo database, of articles published in the last 10 years. Search terms used included methemoglobinemia, cytochrome b5 reductase deficiency, cyanosis, and co-oximetry. Analysis and synthesis of information: Methemoglobinemia can be classified into congenital and acquired, the latter being the most common. It is important to diagnose this disease, which, although it is a rare condition, can cause serious complications, and even death, which are avoidable with early diagnosis and timely treatment that reduce the complications associated with this condition. Conclusions: The diagnosis and treatment, prophylactic and therapeutic, of methemoglobinemia, in its acute or maintenance stage, require adequate updating of the health professional(AU)

First report of Babesia bigemina in Lama glama in a zoological garden of Veracruz, Mexico.

Zoological gardens represent specialised centres for the preservation of biological inventories and genetic diversity, allowing the recognition of multiple species in critical conservation categories. However, the close coexistence of multiple species of vertebrates that may be associated with various species of ectoparasites may be the cause of the transmission of multiple infectious agents, among which tick-borne pathogens stand out. In these areas, several animal species usually live in a small space and proximity to other wildlife, visitors and keepers. In Mexico, little is known about the disease agents transmitted by arthropods in zoological gardens. For this reason, the aim of this study was to identify the presence of Babesia/Theileria in animals maintained in captivity. As a part of a project identifying vector-borne pathogens in wildlife, 24 animals were sampled in the Miguel Angel de Quevedo zoo. Molecular identification of Babesia/Theileria was realised through amplification of a fragment of the mitochondrial cytB gene and the ribosomal 18S-rDNA. Two neotropical camelids (Lama glama) tested positive (2/3 = 66.6%) to Babesia bigemina. Our results represent the first record of B. bigemina in animals in captivity in a zoological garden in Mexico and the first finding of this haemoparasite in neotropical camelids in Mexico.

Iron Deprivation Modulates the Exoproteome in Paracoccidioides brasiliensis.

Fungi of the Paracoccidioides genus are the etiological agents of the systemic mycosis paracoccidioidomycosis and, when in the host, they find a challenging environment that is scarce in nutrients and micronutrients, such as Fe, which is indispensable for the survival of the pathogen. Previous studies have shown that fungi of this genus, in response to Fe deprivation, are able to synthesize and capture siderophores (Fe3+ chelators), use Fe-containing host proteins as a source of the metal, and use a non-canonical reductive pathway for Fe3+ assimilation. Despite all of these findings, there are still gaps that need to be filled in the pathogen response to metal deprivation. To contribute to the knowledge related to this subject, we obtained the exoproteome of Paracoccidioides brasiliensis (Pb18) undergoing Fe deprivation and by nanoUPLC-MSE. One hundred forty-one proteins were identified, and out of these, 64 proteins were predicted to be secreted. We also identified the regulation of several virulence factors. Among the results, we highlight Cyb5 as a secreted molecule of Paracoccidioides in the exoproteome obtained during Fe deprivation. Cyb5 is described as necessary for the Fe deprivation response of Saccharomyces cerevisiae and Aspergillus fumigatus. Experimental data and molecular modeling indicated that Cyb5 can bind to Fe ions in vitro, suggesting that it can be relevant in the arsenal of molecules related to iron homeostasis in P. brasiliensis.

Gene expression changes in Epinephelus marginatus (Teleostei, Serranidae) liver reveals candidate molecular biomarker of iron ore contamination.

Wastes from iron ore mining activities are potentially damaging to adjacent aquatic ecosystems. We aimed to determine biomarkers of environmental exposure to this xenobiotic in the dusky grouper Epinephelus marginatus by differential gene expression analysis. For this, fish were exposed to iron ore (15.2 mg/L) and gene expression in liver was assessed by RNA-Seq and compared to the control group. A total of 124 differentially expressed genes were identified, from which 52 were upregulated and 72 were downregulated in response to iron ore. From these, ferritin (medium subunit), cytochrome b reductase and epoxide hydrolase genes were selected for validation by RT-qPCR that confirmed the upregulation of epoxide hydrolase in fish exposed to iron ore.

Two new species of Peromyscus (Cricetidae: Neotominae) from the Transverse Volcanic Belt of Mexico.

Specimens of the Peromyscus boylii species group distributed in the western and northeastern montane regions of Michoacán, México, historically have been assigned to P. levipes. Previous studies indicated that these specimens possessed mitochondrial DNA haplotypes that were distinct from both P. levipes and P. kilpatricki, a recently named species in the P. boylii species group from northeastern Michoacán and western Morelos. Herein karyotypic, DNA sequence, and morphological data were analyzed from those populations to evaluate their taxonomic affinity. Karyotypic data indicated that individuals from western Michoacán (Dos Aguas and Aguililla) and from a newly discovered population in northeastern Michoacán (Zinapécuaro) were chromosomally similar to P. carletoni (FN = 68) but distinct from other taxa assigned to the P. boylii species group. Analyses of cranial characteristics indicated that, relative to other species in the P. boylii species group, two morphologically distinct groups were present that corresponded to the Dos Aguas/Aguililla and Zinapécuaro populations, respectively. The latter population, although represented by a small sample size (n = 5 specimens), appeared to exhibit some trenchant morphological distinctions compared with other cryptic species in the P. boylii group. Phylogenetic analyses (parsimony, Bayesian, and likelihood) of DNA sequences obtained from the mitochondrial cytochrome-b gene indicated that although the individuals from Dos Aguas/Aguililla and Zinapécuaro formed a sister group relationship, they formed monophyletic clades that differed genetically (2.54%)-a level approaching that seen between other sister species of Peromyscus. Further, the Dos Aguas/Aguililla and Zinapécuaro clade was more closely aligned with a clade containing representatives of P. carletoni and P. levipes instead of with those from closer geographic proximities (P. kilpatricki) located in eastern Michoacán. Together, these results indicated that these two populations seemingly represent two undescribed species in the P. boylii species group for which we propose the names Peromyscus greenbaumi for populations in western Michoacán (circa Dos Aguas and Aguililla) and Peromyscus ensinki for populations in northeastern Michoacán (circa Zinapécuaro).

Los especímenes del grupo de especies de Peromyscus boylii distribuidos en las regiones montañosas occidentales y el noreste de Michoacán, México, históricamente fueron asignados a P. levipes. Sin embargo, estudios previos han indicado que estos especímenes poseen haplotipos de ADN mitocondrial que son distintos de P. levipes y P. kilpatricki, una especie recientemente nombrada en el grupo de especies P. boylii del noreste de Michoacán y el oeste de Morelos. Los datos cariotípicos indicaron que los individuos del oeste de Michoacán (Dos Aguas y Aguililla) y de una población recién descubierta en el noreste de Michoacán (Zinapécuaro) eran cromosómicamente similares a P. carletoni (FN = 68), pero distintos de otros taxones asignados al grupo de especies P. boylii. Los análisis de las características craneales indicaron que en relación con otras especies del grupo de especies P. boylii, dos grupos morfológicamente distintos estaban presentes, y que correspondían a las poblaciones de Dos Aguas/Aguililla y Zinapécuaro, respectivamente. Aunque representada por un tamaño de muestra pequeño (n = 5 especímenes), esta última población pareció exhibir algunas diferencias morfológicas en comparación con las otras especies crípticas en el grupo P. boylii. Los análisis filogenéticos (parsimonia, inferencia bayesiana y verosimilitud) de secuencias de ADN obtenidas del gen mitocondrial citocromo-b, indicaron que, aunque los individuos de Dos Aguas/Aguililla y Zinapécuaro tienen una relación de grupo hermano, estos forman clados monofiléticos que difieren genéticamente (2.54%), nivel que se acerca al observado entre otras especies hermanas de Peromyscus. Además, el clado de Dos Aguas/Aguililla y Zinapécuaro está más estrechamente alineado con un clado que contenía representantes de P. carletoni y P. levipes en lugar de aquellos de proximidades geográficas más cercanas (P. kilpatricki) ubicados en el este de Michoacán. Estos resultados indicaron que estas dos poblaciones aparentemente representan dos especies no descritas en el grupo de especies P. boylii para las cuales proponemos los nombres Peromyscus greenbaumi para poblaciones en el oeste de Michoacán (hacia Dos Aguas y Aguililla) y Peromyscus ensinki para poblaciones en el noreste de Michoacán (circa Zinapécuaro).

International interlaboratory validation of a nested PCR for molecular detection of Babesia bovis and Babesia bigemina, causative agents of bovine babesiosis.

Babesia bovis and B. bigemina are tick-transmitted parasites causing bovine babesiosis, characterized by significant morbidity and mortality leading to economic losses to the livestock industry in tropical and subtropical regions worldwide. Animals that recover from acute infection remain carriers with low parasitemia acting as a source of transmission, and often escape detection. An improved diagnosis of a B. bovis and/or B. bigemina infection of carrier animals is enabled by the availability of detection methods with high sensitivity. To this end, two nested PCR assays targeting the cytochrome b (cytb) genes of B. bovis and B. bigemina (cytb-nPCR), have been recently developed and an increased sensitivity with respect to reference protocols has been shown (Romero-Salas et al., 2016). In this study, the specificity against a panel of hemoparasites that potentially co-occur with B. bovis and B. bigemina was demonstrated to ensure applicability of the cytb-nPCR assays in a wide range of regions where bovine babesiosis is endemic. Furthermore, we compared both reported cytb-nPCR assays with reference nPCR and qPCR protocols for (i) their capability to detect carrier animals in the field, and (ii) their reproducibility when performed in different laboratories by independent operators. We show that, in a panel of bovine field samples (n = 100), the cytb-nPCR assays detected a considerably higher number of 25% B. bovis and 61% B. bigemina-positive animals compared to 7% and 20% B. bovis and 55% and 49% B. bigemina-positive animals when tested by reference nPCR and qPCR protocols, respectively. Cytb-nPCRs were also found superior in the detection of carrier animals when field samples from Africa were analyzed. In addition, both the B. bovis and B. bigemina cytb-nPCR assays were independently validated in a single blinded study in three laboratories. Importantly, no significant differences in the number/percentage of infected animals was observed using cytb-nPCR assays. In summary, the cytb-nPCR assays detected a considerably higher number of chronically infected B. bovis and B. bigemina carrier animals compared to reference nPCR and qPCR protocols, when applied in different epidemiological field situations. Furthermore, a high reproducibility between laboratories could be demonstrated.

Diagnostic locus for identification of silverside hybrids (Odontesthes humensis and Odontesthes bonariensis) in a natural population / Locus diagnóstico para identificação de híbridos de peixe-rei (Odontesthes humensis and Odontesthes bonariensis) em população natural

Hybridization is a natural phenomenon that occurs more often in fish than in other vertebrates. The use of nuclear and mitochondrial molecular markers provides valuable results in the detection of these events. The aim of this study was to investigate the occurrence of interspecific hybrids in natural populations of silverside. The samples of Odontesthes humensis, Odontesthes bonariensis, and indivi-duals that were morphologically different from pure species were collected in the Mangueira lagoon, located in southern Brazil. Result: Six tetranucleotide microsatellite loci were synthesized and tested. The UFPEL_OH3 locus proved to be diagnostic for the detection of silverside hybrids, and it was possi-ble to distinguish between pure and hybrid species. The mitochondrial marker gene cytb synthesized from conserved Odontesthes sequences in the GenBank genetic database showed no differences in the genetic sequence of the samples, needing further studies to confirm the hypothesis.(AU)

A hibridação é um fenômeno natural que ocorre mais frequentemente em peixes do que em outros vertebrados. Para detectá-la, são utilizados marcadores moleculares nucleares e mitocondriais, os quais fornecem resultados valiosos. O objetivo deste estudo foi investigar a ocorrência de híbridos in-terespecíficos em populações naturais de peixe-rei. As amostras de Odontesthes humensis, Odontesthes bonariensis e de indivíduos morfologicamente diferentes das espécies puras foram coletadas na lagoa Mangueira, localizada no Sul do Brasil. Foram sintetizados e testados seis loci microssatélites tetranu-cleotídeos. O locus UFPEL_OH3 mostrou-se um diagnóstico para detectar híbridos de peixe-rei, pos-sibilitando a distinção de espécies puras de híbridas. O marcador mitocondrial gene cytb, sintetizado com base nas sequências conservadas de Odontesthes no banco de dados genéticos do GenBank, não apresentou diferenças na sequência genética das amostras, portanto, são necessários mais estudos para confirmar a hipótese de hibridação.(AU)