First report of Irpex rosettiformis causing white root rot in avocado trees in Michoacán, México.

In México, avocado production is an important economic source. In the last season it generated $ 3. 27 billion USD of foreign currency in the country. Irpex spp. are wood decay fungi. In the period 2019-2022, in the state of Michoacán (19°13' N; 101°55' W), México, basidiomes of Irpex sp. were observed on the base of trunks and crowns of 5-years-old and older avocado (Persea americana) trees. The trees exhibited disease symptoms that included white root rot, leaf yellowing, small leaves, branch diebacks, generalized defoliation, apical flaccidity, abundant but small sun burnt fruits due to the lack of foliage, and after 2-4 years of first disease appearance, the infected trees died. In the place where fungus was established, abundant white and cottony mycelium was formed, which caused trees decay. The incidence of the disease in the sampled orchards was estimated to be 30% per ha with 350 - 400 trees, which was determined through a simple sampling design focused on trees with signs and symptoms of the disease due to the phytopathogen. Samples of infected tissue (roots and stems) and fungal basidiomes were collected from 90 trees (5-6 per orchard). The symptomatic avocado trees studied were randomly selected from 17 orchards. For the fungal macroscopic characterization, the synoptic keys described by Gilbertson and Ryvarden (1986) and by Largent (1973) were used. The samples showed typical structures corresponding to Irpex sp., including rosettes, annual basidiomes, a system of monomitic hyphae, and subglobose basidiospores. In vitro fungal isolation from basidiomes and infected tree tissues was done according to the protocol of Agrios (2004). The fungal strains were maintained on PDA at 28 °C. At 16 days of incubation the colonies were opaque, whitish with fluffy and corky mycelium. Microscopic analysis of the fungus showed typical yellowish spores, with an ellipsoid shape of 3-4 x 4-5.5 µm (50 accounted structures per isolate [N=19]) and basidia of 20-25 x 4.5-5.5 µm (n=20 basidiomes). For molecular characterization, two molecular markers were used, the internal transcribed spacer rDNA-ITS1 5.8 rDNA-ITS2 (ITS; White et al. 1990) and the large ribosomal subunit (LSU; Vilgalys and Hester 1990). The PCR reaction was performed as described by Martínez-González et al. (2017). The consensus sequences were compared with those deposited in the NCBI-GenBank, using the BLASTN 2.2.19 tool (Zhang et al. 2000), the samples showed 99% match with the species, Irpex rosettiformis. GenBank accession numbers of the submitted isolates are summarized in supplementary Table 4. To test Koch's postulates, 3-months old avocado plants grown in greenhouse conditions were inoculated (n = 10 per each isolate [N= 19]) on the roots with 3 g of I. rosettiformis mycelium. The experiment was done twice with 20 non-inoculated plants as control. After 67 days, basidiomes (50 x 70 x 1.5 mm in average) were observed where the disease incidence was >77%, with subsequent tree decline. The pathogen was re-isolated in vitro in PDA and its identity was confirmed by morphological characteristics of mycelium. This work shows that I. rosettiformis is not only a wood decay fungus, but also a phytopathogen, the causative agent of white root rot disease in P. americana var. drymifolia, cultivar 'Hass', which establishes a precedent for monitoring and preventing its proliferation to other regions in the American continent and the world where nursery avocado seedlings are exported.

Berry species and crop management approaches affect species diversity and abundance of predatory mites (Acari: Phytoseiidae).

Berries comprise an economically important group of crops. Knowledge about their arthropod pests and biological control agents is important in the development of more efficient integrated pest management programs. Identification of potential biocontrol agents based solely on morphological attributes may be difficult and so molecular techniques should be incorporated. Here we studied the species diversity of predatory mites in the family Phytoseiidae, and how this diversity is affected by the berry species and crop management approaches, specifically pesticide application regimes. We sampled 15 orchards in the State of Michoacán, Mexico. Sites were selected based on berry species and pesticide regimes. Mite identification was achieved by combining morphological attributes and molecular techniques. Phytoseiidae diversity was compared amongst blackberry, raspberry and blueberry. Subsequently we studied the effect of berry species and pesticide regime on the abundance of the most prevalent phytoseiid species. We identified 11 species of phytoseiid mites. The greatest species diversity was found in raspberry, followed by blackberry and then blueberry. The most abundant species were Typhlodromalus peregrinus and Neoseiulus californicus. The abundance of T. peregrinus was significantly affected by pesticide application but not by berry species. In contrast, abundance of N. californicus was significantly affected by berry species but not by pesticide regime.

Deforestation increases the abundance of rodents and their ectoparasites in the Lacandon forest, Southern Mexico

Introduction: Tropical forests provide important ecosystem services, including disease control. However, few studies have focused on how deforestation affects species more suitable to be zoonotic vectors. Objective: To evaluate how deforestation affects the abundance and species richness of rodents and their associated ectoparasites in a tropical ecosystem. Methods: We captured rodents in 6 landscape units, 1 km² each, with 0.7; 5; 40; 46; 78 and 95 % tree cover, in Marques de Comillas, Chiapas, Southern Mexico. In each unit we set 90 Sherman traps that were active 24 hours for 7 days during two sampling seasons (October 2019, and September 2020). We manually extracted ectoparasites from all captured rodents. Results: We captured 70 rodents of five species: Sigmodon toltecus, Heteromys desmarestianus, Ototylomys phyllotis, Peromyscus mexicanus, and Oryzomys couesi. Rodent abundance increased with forest loss (R²= 0.706, P= 0.022). The greatest richness of rodent species occurred in sites with intermediate forest cover (40 and 78 %). The most abundant species were: S. toltecus (N= 45) followed by O. couesi (N= 9), these species dominated in sites with less forest cover. We recorded a total of 23 ectoparasite species, three of them known to be zoonotic vectors: Amblyomma sp., Ornithonyssus bacoti, and Androlaelaps fahrenholzi. Conclusions: The ongoing loss of forests promotes the proliferation of zoonotic disease vectors in this tropical ecosystem, which can potentially increase the frequency of affectation among the local population.

Introducción: Un servicio particularmente importante que brindan los bosques tropicales es el control de enfermedades. Sin embargo, pocos estudios se han enfocado en analizar cómo este servicio es afectado por la deforestación. Objetivo: Evaluar el efecto de la deforestación en la abundancia y riqueza de especies de roedores y de sus ectoparásitos en Marqués de Comillas, en el sureste de México. Métodos: Capturamos roedores en 6 unidades del paisaje (UP), cada una de 1 km², con distintos porcentajes de cobertura vegetal (0.7, 5, 40, 46, 78 y 95 %). En cada UP colocamos 90 trampas Sherman, que permanecieron activas las 24 horas por 7 días durante dos muestreos en octubre 2019 y septiembre 2020. Todos los roedores capturados fueron revisados para detectar ectoparásitos en su pelaje que fueron recolectados para su posterior identificación en el laboratorio. Resultados: Capturamos 70 roedores de cinco especies: Sigmodon toltecus, Heteromys desmarestianus, Ototylomys phyllotis, Peromyscus mexicanus y Oryzomys couesi. La abundancia de roedores aumentó con la pérdida de bosque (R² = 0.706, P = 0.022). La mayor riqueza de especies de roedores se presentó en sitios con cobertura forestal intermedia (40 y 78 %). Las especies más abundantes fueron: S. toltecus (N = 45) seguido de O. couesi (N = 9), estas especies dominaron en los sitios con menor cobertura forestal. Registramos un total de 23 ectoparásitos diferentes, identificamos 15 a nivel de especie y ocho a nivel de género. Los sitios con menor cobertura forestal presentaron la menor riqueza de especies de ectoparásitos. Detectamos tres especies de ectoparásitos (Amblyomma sp., Ornithonyssus bacoti y Androlaelaps fahrenholzi) que se sabe que son vectores de enfermedades zoonóticas. Conclusión: Encontramos que la deforestación está promoviendo un aumento en la proliferación de vectores de enfermedades zoonóticas lo que, a su vez, tiene el potencial de incrementar las afectaciones de la población local.

Tick-borne rickettsial pathogens in questing ticks, removed from humans and animals in Mexico.

Tick-borne rickettsial diseases (TBRD) are commonly encountered in medical and veterinary clinical settings. The control of these diseases is difficult, requiring disruption of a complex transmission chain involving a vertebrate host and ticks. The geographical distribution of the diseases is related to distribution of the vector, which is an indicator of risk for the population. A total of 1107 were collected by tick drag from forests, ecotourism parks and hosts at 101 sites in 22 of the 32 states of Mexico. Collected ticks were placed in 1.5 mL cryovials containing 70% ethanol and were identified to species. Ticks were pooled according to location/host of collection, date of collection, sex, and stage of development. A total of 51 ticks were assayed by polymerase chain reaction (PCR) to confirm species identification using morphological methods. A total of 477 pools of ticks were assayed using PCR techniques for selected tick-borne pathogens. Anaplasma phagocytophilum was the most commonly detected pathogen (45 pools), followed by, Ehrlichia (E.) canis (42), Rickettsia (R.) rickettsii (11), E. chaffeensis (8), and R. amblyommii (1). Rhipicephalus sanguineus was the tick most frequently positive for selected pathogens. Overall, our results indicate that potential tick vectors positive for rickettsial pathogens are distributed throughout the area surveyed in Mexico.

Implications of climate change on the distribution of the tick vector Ixodes scapularis and risk for Lyme disease in the Texas-Mexico transboundary region.

BACKGROUND: Disease risk maps are important tools that help ascertain the likelihood of exposure to specific infectious agents. Understanding how climate change may affect the suitability of habitats for ticks will improve the accuracy of risk maps of tick-borne pathogen transmission in humans and domestic animal populations. Lyme disease (LD) is the most prevalent arthropod borne disease in the US and Europe. The bacterium Borrelia burgdorferi causes LD and it is transmitted to humans and other mammalian hosts through the bite of infected Ixodes ticks. LD risk maps in the transboundary region between the U.S. and Mexico are lacking. Moreover, none of the published studies that evaluated the effect of climate change in the spatial and temporal distribution of I. scapularis have focused on this region. METHODS: The area of study included Texas and a portion of northeast Mexico. This area is referred herein as the Texas-Mexico transboundary region. Tick samples were obtained from various vertebrate hosts in the region under study. Ticks identified as I. scapularis were processed to obtain DNA and to determine if they were infected with B. burgdorferi using PCR. A maximum entropy approach (MAXENT) was used to forecast the present and future (2050) distribution of B. burgdorferi-infected I. scapularis in the Texas-Mexico transboundary region by correlating geographic data with climatic variables. RESULTS: Of the 1235 tick samples collected, 109 were identified as I. scapularis. Infection with B. burgdorferi was detected in 45% of the I. scapularis ticks collected. The model presented here indicates a wide distribution for I. scapularis, with higher probability of occurrence along the Gulf of Mexico coast. Results of the modeling approach applied predict that habitat suitable for the distribution of I. scapularis in the Texas-Mexico transboundary region will remain relatively stable until 2050. CONCLUSIONS: The Texas-Mexico transboundary region appears to be part of a continuum in the pathogenic landscape of LD. Forecasting based on climate trends provides a tool to adapt strategies in the near future to mitigate the impact of LD related to its distribution and risk for transmission to human populations in the Mexico-US transboundary region.

First phylogenetic analysis of Ehrlichia canis in dogs and ticks from Mexico. Preliminary study / Primer análisis filogenético de Ehrlichia canis en perros y garrapatas de México. Estudio preliminar

ABSTRACT Objective. Phylogenetic characterization of Ehrlichia canis in dogs naturally infected and ticks, diagnosed by PCR and sequencing of 16SrRNA gene; compare different isolates found in American countries. Materials and methods. Were collected Blood samples from 139 dogs with suggestive clinical manifestations of this disease and they were infested with ticks; part of 16SrRNA gene was sequenced and aligned, with 17 sequences reported in American countries. Two phylogenetic trees were constructed using the Maximum likelihood method, and Maximum parsimony. Results. They were positive to E. canis 25/139 (18.0%) dogs and 29/139 (20.9%) ticks. The clinical manifestations presented were fever, fatigue, depression and vomiting. Rhipicephalus sanguineus Dermacentor variabilis and Haemaphysalis leporis-palustris ticks were positive for E. canis. Phylogenetic analysis showed that the sequences of dogs and ticks in Mexico form a third group diverging of sequences from South America and USA. Conclusions. This is the first phylogenetic analysis of E. canis in Mexico. There are differences in the sequences of Mexico with those reported in South America and USA. This research lays the foundation for further study of genetic variability.

RESUMEN Objetivos. Caracterizar filogenéticamente Ehrlichia canis a partir de perros naturalmente infectados y sus garrapatas, mediante PCR y secuenciación del gene 16SrRNA para compararlos con diferentes aislados encontrados en el continente Americano. Material y métodos. Se colectaron muestras sanguíneas de 139 perros con manifestaciones clínicas sugestivas a Ehrlichiosis, y que estuvieran infestados con garrapatas; una parte del gene 16SrRNA, fue secuenciada y alineada junto con las 17 secuencias reportadas en los países de América. Se construyeron dos árboles filogenéticos utilizando el método de Máxima verosimilitud compuesta, y Máxima parsimonia. Resultados. Fueron positivos a E. canis 25/139 (18.0%) perros y 29/139 (20.9%) garrapatas colectadas sobre los perros. Las manifestaciones clínicas presentadas fueron fiebre, astenia, depresión y vómito. Las garrapatas Rhipicephalus sanguineus, Dermacentor variabilis y Haemaphysalis leporis-palustris fueron positivas para E. canis. El análisis filogenético mostró que las secuencias 16SrRNA de Ehrlichia canis aisladas de perros y garrapatas en este estudio forman un tercer grupo que diverge de las secuencias de Sudamérica y EUA. Conclusiones. Es el primer análisis filogenético de E. canis en México. Hay diferencias entre las secuencias de este estudio, con las reportadas en otros países de Sudamérica y en EUA. Esta investigación sienta las bases para profundizar en el estudio de la variabilidad genética.